Skip to main content
NCBI home page
The Journal of Venomous Animals and Toxins Including Tropical Diseases logoLink to The Journal of Venomous Animals and Toxins Including Tropical Diseases
. 2025 Sep 12;31:e20250007. doi: 10.1590/1678-9199-JVATITD-2025-0007

Proteasome-driven modulation of immune and oxidative pathways during scorpion envenomation pathogenesis

Amal Megdad-Lamraoui 1, Sonia Adi-Bessalem 1, Fares Daachi 1, Fatima Laraba-Djebari 1,2,*
PMCID: PMC12431687  PMID: 40951511

Abstract

Background:

Scorpion venom contains a variety of toxin molecules that are the drivers of inflammation and oxidative stress, leading to significant tissue damage. While several mechanisms underlying these responses have been studied, the involvement of the proteasome complex - a key regulator of inflammation - remains poorly understood. This study explored the role of the proteasome in modulating inflammatory and oxidative responses to envenomation by Androctonus australis hector venom.

Methods:

Mice were pretreated intraperitoneally with bortezomib, a proteasome inhibitor, at low (0.05 mg/kg), medium (0.25 mg/kg), or high (0.5 mg/kg) doses, 30 minutes prior to sublethal venom administration (0.5 mg/kg, subcutaneous). Twenty-four hours after venom administration, animals were euthanized, blood and organs were collected to evaluate vascular permeability (via Evans blue dye extravasation), the extent of inflammatory cell infiltration (myeloperoxidase and eosinophil peroxidase enzymatic activities), and oxidative/nitrosative stress markers (nitric oxide, hydrogen peroxide, malondialdehyde, catalase activity, and glutathione). Histopathological examinations were performed to identify structural alterations, such as edema, hemorrhage, and cellular infiltration. Biochemical parameters reflecting organ function, including serum levels of CPK, LDH, ALT, ALP, urea, and creatinine, were also measured to assess the degree of systemic damage.

Results:

Our findings revealed a dose-dependent immune-modulatory role of the proteasome system. A medium dose of bortezomib reduced inflammatory and oxidative stress markers, such as vascular permeability, eosinophil peroxidase, neutrophil peroxidase, nitric oxide, and malondialdehyde in renal tissue, suggesting a reduction in local inflammation and oxidative damage. In contrast, a higher dose showed pronounced preventive effects in cardiopulmonary and hepatic tissues, significantly reducing inflammatory mediators and oxidative markers, restoring antioxidant enzyme activity (catalase) and glutathione, as well as, improving tissue structure and organ function.

Conclusion:

These findings underscore the proteasome involvement in inflammatory regulation, likely through modulation of vascular permeability, immune cell activation, and oxidative stress, making it a key target in scorpion envenomation.

Keywords: Proteasome, Scorpion venom, Cardiopulmonary tissue, Hepatorenal tissue, Immune-inflammatory response, Oxidative stress

Background

Despite several scientific efforts and many therapeutic advances within the last decades, scorpion stings are still a real threat that remains among the causes of death especially in children and elderly persons in tropical and subtropical regions of the world [1-3]. Scorpion envenomation generates various signs ranging from local effects to systemic manifestations including neurotoxicity, nephrotoxicity, cytotoxicity, allergic reactions, cardiopulmonary and hepatorenal system dysfunction [4-9]. Scorpion venom contains short and long chain neurotoxic peptides that modulate the function of ion channels in neuronal terminals and are usually responsible for the main symptoms of envenoming [10, 11]. Besides these neurotoxic actions, venom components can also induce harmful inflammatory effects and excessive production of inflammatory mediators by interacting with innate immunity receptors (TLRs) and involving different inflammatory pathways [12-17].

Previous studies have demonstrated the involvement of several systems in the occurrence of inflammatory pathophysiological disturbances during scorpion envenomation - such as the adrenergic, cholinergic, histaminergic, kinin-kallikrein, renin-angiotensin-aldosterone as well as the complement system - inducing the release of different mediators including histamine, prostaglandins, leukotrienes and cytokines [6, 8, 11, 13, 16, 18-24]. Oxidative stress damage caused by excessive reactive oxygen species (ROS) is also one of the common insults encountered by cells during scorpion envenomation altering tissue integrity [25-29]. It has been shown in various types of cells that modified proteins by oxidation are selectively hydrolyzed by proteolytic systems [30, 31]. The proteasome is the main proteolytic system which is not only involved in abnormal protein degradation, it also plays a key role in several other fundamental cellular processes including the regulation of protein homeostasis, major histocompatibility (MHC) class I antigen processing, cell cycle proliferation, signaling and modulation of the immune and inflammatory responses [32-34].

The proteasome system is also known to be involved in regulation of the nuclear factor-κB (NF-κB) activity, which controls the transcription of a wide range of genes responsible for inflammation, such as cytokines, adhesion molecules (ICAM-1, VCAM-1), inducible nitric oxide synthase (iNOS), cyclo-oxygenase 2 (COX-2) and stress-response proteins [35-37].

The involvement of the proteasome system in the development of inflammation response and oxidative stress during scorpion envenomation pathogenesis has not been investigated. The overall objective of this study was to clarify the immune modulatory role of the proteasome system in peripheral tissues during scorpion envenomation.

Methods

Venom

Lyophilized venom of Androctonus australis hector (Aah) was obtained from the Biochemistry of Biomolecules: Mode of Action, Immunotherapy and Immunodiagnosis team, Laboratory of Cellular and Molecular Biology, Faculty of Biological Sciences, University of Sciences and Technology Houari Boumediene (USTHB). A previous study reported that the median lethal dose (LD₅₀) of this venom is approximately 0.85 mg/kg when administered via the intraperitoneal (i.p.) route [38]. In contrast, our prior work using subcutaneous injection - a route that closely simulates natural envenomation - demonstrated that a dose of 0.5 mg/kg induces mild envenomation without resulting in mortality [6-8, 13, 23, 24, 39].

Chemicals and drugs

Bortezomib (Velcade®, PS-341) provided by Janssen-Cilag (France), is used as a proteasome inhibitor. The chemicals and reagents were mainly from Sigma (St. Louis, MO, USA) and Merck (Mannheim, Germany).

Animals and experimental design procedure

The experiments were performed in Algeria using Naval Medical Research Institute (NMRI) Swiss albino male mice weighing 20-22 g (7-8 weeks old) that did not undergo previous procedures. They were obtained from the animal breeding center of the Faculty of Biological Sciences, FBS-USTHB (Algiers, Algeria). They were housed under a 12-hour light/dark cycle and fed standard pellet chow and tap water ad libitum. All animal experiments were performed according to the Guide for the Care and Use of Laboratory Animals. All animal procedures were conducted in compliance with the ethical standards set forth by the European Parliament and Council Directive on the protection of animals used for scientific purposes (Directive 2010/63/EU). This study was approved by the Deontology and Ethic Committee of the Research Thematic Agency in Health Sciences (ATRSS) formerly National Agency of Research Development in Health (ANDRS).

In order to study the immune-modulatory role of proteasome in peripheral tissues during scorpion envenomation, the animals were randomly divided into five groups of 12 mice each. They were treated as follows: the first group serving as control received an injection of 100 μL/mouse of physiological saline water (NaCl) at 0.9% by subcutaneous route (s.c). The second group represents envenomed mice with a sublethal dose at 0.5 mg/kg (s.c.), while the third, the fourth, the fifth and the sixth groups consist of mice receiving three doses of bortezomib (0.05, 0.25, 0.5 mg/kg) tested intraperitoneally. About 30 minutes later, mice were envenomed with a sublethal dose of Aah venom (0.5 mg/kg, s.c.). Control and envenomed mice were euthanized 24 hours after the injection of physiological saline or venom. In addition to blood, the heart, lungs, liver, kidneys (from control or treated animals with venom at 0.5 mg/kg) were collected 24 hours after NaCl or venom injection, weighed, and then used for further investigations.

Enumeration of the different peripheral blood leukocyte populations

Blood samples were taken in tubes with EDTA 24 hours after the injection of Aah venom. The cell count was carried out by a hemocytometer ERMA INC (full automatic blood cell counter model PCE-210N) analyzer. Blood cells were identified according to the position of the nucleus, its area and its density. The results are expressed as 103 cells/μL of blood.

Serum protein electrophoresis

Serum protein electrophoresis was carried out using an automatic device of the “Capillary SEBIA” type. Serum proteins (albumin and α1, α2, β, γ globulins) are amphoteric molecules, their separation takes place according to their electrical charges. The serum samples from the different groups were placed in a basic medium (cellulose acetate) where the serum proteins acquired an overall negative charge allowing them to migrate from the cathode to the anode under the influence of an electric field. The migration was stopped as soon as the separation was sufficient; a fixation was then carried out with the acid blue dye. Five zones corresponding to the different serum proteins having decreasing negative charges were obtained. The results were expressed in g/L.

Vascular permeability and inflammatory cell infiltration

Vascular permeability

To measure changes in vascular permeability, Evans blue dye was administered intravenously (20 mg/kg) immediately before the injection of NaCl or venom [40]. The organs - heart, lungs, liver and kidneys - were collected after mouse euthanasia. They were then placed in formamide for the extraction of Evans blue, then incubated at 37°C for 72 hours. Absorbance was read at 620 nm and the results were expressed as the concentration of Evans blue per µg of tissue.

Myeloperoxidase activity

The accumulation and activation of neutrophils were analyzed by measuring myeloperoxidase (MPO) in tissue homogenates according to the method described by Coelho et al. [41]. The oxidation of the chromogenic substrate, O-dianisidine, by MPO was measured by spectrophotometry at 460 nm. A volume of 1 mL of 50 Mm phosphate buffer (pH 6.6) containing O-dianisidine dihydrochloride (0.167 mg/mL) and hydrogen peroxide, was added to 1 mL of biological sample. The oxidation of the chromogenic substrate O-dianisidine by MPO was measured spectrophotometrically at 460 nm Changes in absorbance were recorded over 2minutes. MPO activity was expressed as mM/min/100 mg of tissue or mM/min/ mL of serum, using an extinction coefficient (ε) of 11.3 mM⁻¹·cm⁻¹.

Eosinophil peroxidase activity

The evaluation of the eosinophil peroxidase (EPO) activity, a marker for eosinophil accumulation, was carried out according to the method described by Van Oosterhout et al. [42]. Supernatants (50 µL) were placed in the wells of a microplate with 100 μL of buffer solution [50 mM Tris-HCl, pH 8, containing 20 mg of o-phenylenediamine (OPD) and 10 µL of H₂O₂]. Enzyme activity was assessed by measuring absorbance at 490 nm after 1 hour of incubation at 37°C. Results are expressed as absorbance per 100 mg of tissue.

Oxidative stress markers

Oxidative stress was evaluated by measuring the levels of pro-oxidant (malondialdehyde and nitrites) and antioxidants biomarkers (catalase CAT and glutathione GSH) in tissue homogenates.

Nitric oxide

The level of nitric oxide was determined by measuring nitrites [43]. Nitrite levels were evaluated in the various samples using the Griess method. The samples were deproteinized with TCA (10%) and then incubated volume for volume with Griess reagent (1% sulfanilamide and 0.1% naphthylethylenediamine dihydrochloride in 2.5% phosphoric acid) for 20 min at room temperature. Nitrite levels were measured spectrophotometrically at 540 nm. A standard curve was prepared using a stock solution of KNO2. Nitrite concentrations are expressed in µM/g of tissue.

Hydrogen peroxide

The estimation of hydrogen peroxide (H2O2) levels is based on the oxidation of phenol red by H2O2 via peroxidase [44]. The samples were distributed in a microplate at a rate of 100 µL/well supplemented with 100 µL of a reactive solution of phenol red [0.01 g glucose, 0.0001 g of horseradish peroxidase (HRP) 0.0001 g phenol red in 10 mL PBS]. The plate was then incubated for one hour at 37°C, protected from light. The reaction was stopped by adding 10 µL of NaOH (1N). Absorbance readings were taken at 620 nm microplate reader. The concentration of hydrogen peroxide was determined after extrapolation on a standard curve made for different concentrations of H2O2 ranging from 0.005 mM to 0.500 mM. Results were expressed as mM/100 mg tissue.

Malondialdehyde

Malondialdehyde, an indicator of lipid peroxidation, is one of the end products of the breakdown of polyunsaturated fatty acids, it is released under the effect of free radicals during stress. The malondialdehyde level was evaluated according to the method of Moreno et al. [45]. The samples from the different groups are incubated (v/v) with TCA (35 %) for 1 hour at 4°C for protein precipitation. After centrifugation of the mixture for 10 minutes at 4000 g, 200 μL of the supernatant were mixed with 100 μL of SDS (8.1%), 750 μL of acetic acid (20%), 750 μL of thiobarbituric acid (0.8%) and 200 μL of distilled water. The mixture was then incubated at 100°C for 1 hour, followed by cooling in ice. The optical density was measured at 532 nm and the amount of malondialdehyde formed was calculated using a molar extinction coefficient of 1.56 x 105 M-1 cm-1. Results were expressed in nM/ 100 mg of tissue.

Catalase activity

Catalase is a ubiquitous antioxidant enzyme. It is responsible for the conversion of hydrogen peroxide (H2O2) into water (H2O) and oxygen (O2). The enzymatic reaction is triggered by the addition of 50 μL of each sample and of the substrate (0.2% H2O2) in phosphate buffer (50 mM at pH 7). The kinetics of H2O2 degradation were measured spectrophotometrically at 240 nm for 3 min. One unit of catalase is equal to (2.3/T) (Log A1/A2), where T is the time interval in minutes, A1 the absorbance in the first minute and A2 the absorbance in the second minute [46]. Catalase activity was expressed in U/100 mg of tissue.

Glutathione

Glutathione plays an important role in the detoxification and reduction of both organic and inorganic peroxides, in the presence of glutathione peroxidase (GPx). Its quantification is based on the measurement of the optical absorbance of 2-nitro-5-mercapturic acid, which results from the reduction of 5,5'-dithio-bis-2-nitrobenzoic acid by the (-SH) groups of GSH [47]. To carry out this assay, a volume of 50 µL of sample was mixed with 100 µL of DTNB (5,5'-dithio-bis (2-nitrobenzoic acid) and 100 µL of phosphate buffer (0.1 M and pH 7.4) The absorbance reading was taken at 450 nm after a 30 minute incubation at 37°C. The results were expressed in µM / 100 mg of tissue using a molar extinction coefficient of 13.3 M-1.cm-1

Histopathological analysis

The effects of proteasome inhibitor on Aah venom induced cardiac, pulmonary, and hepatorenal tissues alteration were examined by histological analysis.

Organs (liver, kidneys, heart, lungs) were collected from the animals, fixed in formalin (10%), and embedded in paraffin. The resulting tissue blocks were sectioned using a microtome to obtain 5 µm thick sections. These sections were stained with hematoxylin and eosin to visualize tissue damage under a light microscope.

Evaluation of organ functions

Organ function was assessed by estimating the enzyme activities of creatine phosphokinase (CPK), lactate dehydrogenase (LDH), alanine aminotransferase (ALT), alkaline phosphatase (ALP), as well as by measuring urea and creatinine levels in serum, according to the manufacturer's (Spinreact, Spain) instructions. Enzyme activities were expressed in international units (IU/L). Serum creatinine and urea levels were expressed in g/L.

Statistical analysis

Results are expressed as means ± SEM. Statistical comparisons were carried out using One-Way ANOVA (Graph Pad Prism 5 Software, San Diego, CA, USA) to identify differences between the envenomed and control groups or between the pretreated and the envenomed groups. A value of p < 0.05 was considered statistically significant.

Results

Effects of proteasome inhibitor on blood cells and serum alpha, beta and gamma globulins concentration

Blood leukocyte levels were assessed in the presence of low (0.05 mg/kg), medium (0.25 mg/kg), or high (0.5 mg/kg) doses of proteasome inhibitor before experimental envenomation. The results showed that scorpion venom induced hyperleukocytosis 24 hours after envenomation of the mice, compared to the control group, as evidenced by a significant increase in the levels of lymphocytes (p < 0.001), monocytes (p < 0.01), and granulocytes (p < 0.05) (Figure 1A). This migration is likely induced by various immune stimuli, such as the pro-inflammatory cytokines released after scorpion envenomation [48-51].

Figure 1. (A) Effects of proteasome inhibition on leukocyte levels and (B) serum protein contents 24 hours after envenomation. Values are expressed as mean ± SEM (n = 3/group). Student's t-test, *p < 0.05; **p < 0.01; ***p < 0.001; ****p <0.0001: groups of envenomed (0.5 mg/kg) mice compared to the control group (NaCl). #p < 0.05; ##p < 0.01; ###p < 0.001; ####p <0.0001: pretreated groups compared to the group of envenomed (0.5 mg/kg) mice. ns: non-significant results.

Figure 1.

Open in a new tab

Serum proteins from the acute phase of inflammation are key markers of the inflammatory response. Electrophoresis analysis revealed an increase in the alpha and gamma globulin fractions in the sera of envenomed mice exposed to a sublethal dose of Aah venom (Figure 1B). Gamma globulins are synthesized following the activation of B lymphocytes. The increase in this fraction indicates the synthesis of various types of immunoglobulins in serum (IgA, IgG, IgM, IgD, IgE) and C-reactive protein (CRP).

Higher doses of proteasome inhibitor (0.25 and 0.5 mg/kg) induced a significant decrease in the level of lymphocytes (p < 0.05 and p < 0.01), monocytes (p < 0.01 and p < 0.001) and granulocytes (p < 0.05 and p < 0.01), comparatively to that of envenomed mice. It seems that this pretreatment was able to significantly prevent hyperleukocytosis induced by Aah venom in the blood (Figure 1A).

With regard to the effects on the serum protein electrophoresis, pretreatment of mice with a low dose of bortezomib (0.05 mg/kg) induced a significant decrease in all globulin fractions (α1, α2, β, and γ) compared to the values recorded in the envenomed group of mice. Bortezomib at 0.25 and 0.5 mg/kg significantly reduced the α1, α2, β- and γ-globulin fractions, compared to the envenomed group of mice. However, a more pronounced decrease in all globulin fractions was observed at the higher concentration of bortezomib (0.5 mg/kg), compared to the effects recorded at the lower doses (Figure 1B).

Effect of pretreatment with proteasome inhibitor on serum albumin level and organ water content

Compared with the control group, the levels of albumin were significantly decreased in envenomed animals (p < 0.0001) (Figure 2A). This hypoalbuminemia is accompanied by high water content in the heart, lungs, liver and kidneys of envenomed animals compared with normal controls. Albumin is a negative acute phase protein. Leakage of serum albumin levels due to increased vascular permeability can contribute to edema formation in tissues [52]. Bortezomib at 0.5 mg/kg significantly prevented the decrease in albumin rates compared with the venom group (p < 0.05). However, no significant effect was observed at lower concentrations (0.05 and 0.25 mg/kg) (Figure 2A). Similarly, with regard to tissue water content, bortezomib at the highest concentration tended to prevent edema formation in the cardiac, pulmonary, kidneys and hepatic tissues (Figure 2B). These results suggest that Aah venom induces edema formation, by activating in part the proteasome complex.

Figure 2. (A) Effects of proteasome inhibition on serum albumin levels and (B) peripheral tissue water content 24 hours after envenomation. Values are expressed as mean ± SEM (n = 3/group). Student's t-test, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001: groups of envenomed (0.5 mg/kg) mice compared to the control group (NaCl). #p < 0.05; ##p < 0.01; ###p < 0.001; ####p < 0.0001: pretreated groups compared to the group of envenomed (0.5 mg/kg) mice. ns: non-significant results.

Figure 2.

Open in a new tab

Evaluation of vascular permeability and inflammatory cell infiltration in peripheral tissues

Vascular permeability in various peripheral tissues was assessed by quantifying Evans blue extravasation [40]. The increase in vascular permeability in damaged tissue reflects the movement of a significant volume of plasma from blood capillaries into the tissues. This plasma flow transports a substantial number of inflammatory mediators [53] leading in particular to edema formation in tissues. Results showed that 24 hours after injection, Aah venom induces a significant increase in vascular permeability of heart (p < 0.0001), lungs (p < 0.0001), liver (p < 0.001) and renal (p < 0.001) tissues (Figure 3A). These results are similar to those carried out previously by our team [6-8, 29].

Figure 3. Effects of low (0.05 mg/kg), medium (0.25 mg.kg) or high dose (0.5 mg/kg) of bortezomib on (A) vascular permeability changes and inflammatory cell infiltration including (B) neutrophils and (C) eosinophils in peripheral tissues of envenomed mice by Aah venom. Values are expressed as mean ± SEM (n = 3/group). Student's t-test, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001: groups of envenomed mice compared to the control group (NaCl). #p < 0.05; ##p < 0.01; ###p < 0.001; ####p < 0.0001: pretreated groups compared to the group of envenomed (0.5 mg/kg) mice. ns: non-significant results.

Figure 3.

Open in a new tab

The increase in vascular permeability facilitates the migration of leukocytes from the vascular system to the injured area. Peroxidase activities were measured as markers of neutrophil and eosinophil activation. Myeloperoxidase and eosinophil peroxidase activities were significantly elevated in the tissue homogenates of animals injected with Aah venom compared to the control group (Figure 3B and 3C).

According to the results, pretreatment with low (0.05 mg/kg), medium (0.25 mg/kg), and high (0.5 mg/kg) doses of proteasome inhibitor significantly reduced Aah venom-induced Evans blue extravasation in cardiac and pulmonary tissues in a concentration-dependent manner. However, in renal tissue, this effect was not dose-dependent, as the highest dose of bortezomib (0.5 mg/kg) failed to prevent the increase in vascular permeability to the same extent as the lower dose (Figure 3A).

The activities of myeloperoxidase and eosinophil peroxidase significantly decreased in the group of animals treated with a higher dose of bortezomib (p < 0.001, p < 0.0001, p < 0.01) compared to the values observed in the heart, lungs, and liver tissues of animals envenomed with Aah venom alone. In the renal tissue it appears that, the medium dose (0.25 mg/kg) of bortezomib is more effective (p < 0.05) than the low and high doses in reducing the infiltration of neutrophils and eosinophils (Figure 3B and 3C).

Evaluation of oxidative stress status in peripheral tissues in the presence or absence of increasing doses of proteasome inhibitor

The inflammatory response induced by Aah venom was associated with a significant increase in nitrites, hydrogen peroxide, and malondialdehyde levels, along with a decrease in glutathione content and catalase activity compared to the controls (Figures 4A, 4B and 4C). Previous studies have suggested that the infiltration of inflammatory cells (including neutrophils and eosinophils) may lead to increased release of reactive oxygen and nitrogen species accompanied by an alteration of the antioxidant defense system [6, 54].

Figure 4. Effects of low (0.05 mg/kg), medium (0.25 mg.kg) or high dose (0.5 mg /kg) of bortezomib on (A) hydrogen peroxide, (B) nitrites and (C) malondialdehyde levels in peripheral tissues of envenomed mice by Aah venom. Values are expressed as mean ± SEM (n = 3/group). Student's t-test, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001: groups of envenomed (0.5 mg/kg) mice compared to the control group (NaCl). # p < 0.05; ## p < 0.01; ### p < 0.001; #### p < 0.0001: pretreated groups compared to the group of envenomed (0.5 mg/kg) mice. ns: non-significant results.

Figure 4.

Open in a new tab

Pretreatment with bortezomib reduced the levels of pro-oxidant markers (MDA, nitrites, H2O2) and prevented dysfunction of the antioxidant defense system (CAT and GSH) in the peripheral tissue homogenates of envenomed mice. The high dose (0.5 mg/kg) appears to be more effective in reducing nitrite and hydrogen peroxide levels, as well as lipid peroxidation, compared to the low and medium doses in all studied tissues, except in renal tissue, when compared to the respective values recorded in envenomed mice without pretreatment (Figures 4A, 4B and 4C). In the renal tissue, significant decrease in pro-oxidant markers were noted in the group receiving a medium-dose (0.25 mg/kg) of bortezomib in comparison with envenomed mice (Figures 4A, 4Band 4C).

The decrease in pro-oxidant markers was associated with an improved antioxidant status, characterized by the restoration of catalase activity and GSH levels in peripheral tissues, compared to the animals injected with Aah venom. Significant results were observed in heart and lung homogenates of mice pretreated with the high dose of bortezomib. At the hepatic level, this dose did not have an effect on the restoration of glutathione levels in the liver. In the kidneys, it appears that the medium and low doses were respectively more effective in preventing alterations in the non-enzymatic (p > 0.05, p < 0.001) and enzymatic (p < 0.05) antioxidant systems (p < 0.0001, p < 0.05) and enzymatic (p < 0.05) (Figure 5A and 5B).

Figure 5. Effects of bortezomib (0.05 mg/kg, 0.25 mg.kg and 0.5 mg /kg) on (A) glutathione and (B) catalase activity in peripheral tissues of envenomed mice by Aah venom. Values are expressed as mean ± SEM (n = 3/group). Student's t-test, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001: groups of envenomed (0.5 mg/kg) mice compared to the control group (NaCl). # p < 0.05; ## p < 0.01; ### p < 0.001; #### p < 0.0001: pretreated groups compared to the group of envenomed (0.5 mg/kg) mice. ns: non-significant results.

Figure 5.

Open in a new tab

Histopathological analysis and the effects of proteasome inhibitor on biomarker levels of organ function in envenomed animals

To further evaluate the role of the proteasome in tissue injuries induced by scorpion venom, histopathological analysis of H&E-stained sections was performed. Histological sections of heart, liver, lung, and kidney tissues from the control group showed normal structure without any histological changes (Figures 6A and 6F, and Figures 7A and 7F). However, scorpion venom administration induced tissue damage, characterized by the loss of normal histological structure, infiltration of inflammatory cells, thickening of the alveolar septa, as well as edema and hemorrhage (Figures 6Band 6G, and Figures 7B and 7G). These histological changes were accompanied by a significant increase in CPK, ALT, ALP, LDH, urea, and creatinine levels in the sera of envenomed mice (Table 1). The increase in these markers may be attributed to impaired organ function, as shown in previous studies [55, 56].

Figure 6. Effects of proteasome inhibition on (A-E) myocardial and (F-J) hepatic tissue, 24 h after Aah envenomation. (A, F) Control (NaCl). (B, G) Animals injected with Aah venom (s.c). (C, H) Animals pretreated with a low dose bortezomib (0.05 mg/kg, i.p.). (D, I) Animals pretreated with a medium dose bortezomib (0.25 mg/kg, i.p.). (E, J) Animals pretreated with a high dose bortezomib (0.5 mg/kg, i.p.). Hematoxylin-eosin staining, magnification × 400. Cv: center-lobe vein; CCv: congested center-lobe vein; Ed: edema; H: hemorrhage; HEd: hemorrhagic edema; Ic: inflammatory cell infiltrates; N: nucleus.

Figure 6.

Open in a new tab

Figure 7. Effects of proteasome inhibition on (A-E) pulmonary and (F-J) renal tissue structure, 24 h after Aah envenomation. (A, F) Control (NaCl). (B, G) Animals injected with Aah venom (s.c). (C, H) Animals pretreated with a low dose bortezomib (0.05 mg/kg, i.p.). (D, I) Animals pretreated with a medium dose bortezomib (0.25 mg/kg, i.p.). (E, J) Animals pretreated with a high dose bortezomib (0.5 mg/kg, i.p.). Hematoxylin-eosin staining, magnification × 400. A: alveolar; DT: distal tubule; E: edema; Gl: glomerulus; HEd: hemorrhagic edema; H: hemorrhage; Ic: inflammatory cell infiltrates; Is: inter-alveolar septum; Nc: necrosis; PT: proximal tubule; TIs: thick inter-alveolar septum.

Figure 7.

Open in a new tab

Table 1. Effects of proteasome inhibition on serum biomarkers of tissue damage. Values represented in the table are expressed as mean ± SEM (n = 3/group). Student's t-test, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001: groups of envenomed (0.5 mg/kg) mice compared to the control group (NaCl). # p < 0.05; ## p < 0.01; ### p < 0.001; #### p < 0.0001: pretreated groups compared to the group of envenomed (0.5 mg/kg) mice. ns: non-significant results.

Control (NaCl) Venom Bortezomib (0.05 mg/kg) + venom Bortezomib (0.25 mg/kg) + venom Bortezomib (0.5 mg/kg) + venom
CPK (IU/L) 1626.66 ± 46.45 2530 ± 107.33*** 2070.33 ± 306.77# 2000.33 ± 100.10# 1605.66 ± 2.88###
ALT (IU/L) 35.83 ± 4.59 170.28 ± 18.25**** 164.16 ± 7.63 ns 132.00 ± 12.58 # 38.33 ± 12.58####
ALP (IU/L) 84.53 ± 9.47 193.89 ± 17.68** 122 ± 31.32# 100 ± 21.32## 102.33 ± 28.29####
LDH (IU/L) 1698.66 ± 34.42 1895.00 ± 107.58 1836.68±60.27ns 1721.66 ± 235.39ns 1503.33 ± 66.16#
Urea (g/L) 0.16 ± 0.04 0.42 ± 0.03**** 0.40 ± 0.01ns 0.26 ± 0.01## 0.37 ± 0.06ns
Creatinine (g/L) 3.33 ± 0.57 11.33 ± 2.3** 8 ± 2.00 ns 7 ± 2.00ns 11 ± 2.64ns
Open in a new tab

Following treatment with a high dose of bortezomib, administered intraperitoneally 30 minutes before envenomation, tissue alterations were significantly reduced in the heart, liver, and lungs (Figures 6E and 6J, and Figure 7E). This pretreatment significantly reduced the activities of CPK (69.28%), ALT (23.63%), ALP (52.37%), and LDH (20.67%) compared to the animals injected with Aah venom (Table 1). The low and medium doses of bortezomib treatment resulted in a slight reduction in histological injuries in these organs. However, some histological alterations, such as edema, inflammatory cell infiltration, and congestion of the central vein, persisted (Figures 6C, 6H, 6D and 6I, and Figures 7Cand 7D).

In the renal tissue, marked reduction of tissue alterations was observed in average-dose (0.25 mg/kg) of bortezomib in pretreated group of mice (Figure 7I) accompanied with significant decrease in urea (73.21%) and creatinine (28.42%) levels when compared to envenomed group of mice (Table 1).

Discussion

Involvement of the proteasome system during various inflammatory diseases was reported and has been directly linked to the progression of these diseases [57-61]. However, no study has yet demonstrated its role in the pathogenesis of scorpion envenomation.

In this study, we investigated the role of the proteasome in Aah venom-induced toxicity and evaluated the potential immune-modulatory role of bortezomib, a proteasome inhibitor, through its impact on vascular, oxidative and biochemical markers.

The present study showed that experimental envenomation with Aah venom triggered systemic immune system activation, characterized by increased blood cell counts and acute-phase protein levels, along with hypoalbuminemia. These findings are consistent with previous studies on scorpion envenomation, which have also reported systemic inflammatory responses, including leukocytosis, elevated acute-phase reactants, and reduced albumin levels [62-64]. The earliest response to envenomation is vascular dysfunction, characterized by increased vascular permeability and tissue edema. This was reflected in our study by elevated tissue water content and Evans blue dye extravasation in multiple organs. Similar results were reported in previous studies on scorpion envenomation [6, 8]. Hypoalbuminemia observed in this context may result from increased vascular permeability, which facilitates the leakage of albumin and other plasma proteins into tissues. Hypoalbuminemia reduces plasma oncotic pressure, leading to fluid leakage into tissues, which results in organ water accumulation, tissue swelling, and potential functional impairment [65]. The vascular leakage is often associated with inflammatory cell infiltration, as confirmed in this study by the elevated activities of myeloperoxidase (MPO) and eosinophil peroxidase (EPO), enzymatic markers of neutrophil and eosinophil accumulation, respectively. These immune cells are known to be major sources of reactive oxygen species, such as hydrogen peroxide (H₂O₂) generated from superoxide anions, as well as reactive nitrogen species like nitric oxide (NO) [66, 67]. The overproduction of nitric oxide (NO) and reactive oxygen species (ROS) during envenomation leads to a significant redox imbalance, partly due to reduced antioxidant defense, as shown by the decreased glutathione level and catalase activity likely inhibited by excess H₂O₂ [68, 69]. These reactive species act as cytotoxic agents by inducing lipid peroxidation, compromising cell membrane integrity [70]. This was evaluated by elevated malondialdehyde (MDA) levels, both in our study and in previous experimental and clinical reports [48, 71], reflecting oxidative tissue damage. Histological analysis further revealed venom-induced cardiac, pulmonary and hepatorenal alterations, including hemorrhage, edema, and inflammatory cell infiltration. These results were also obtained by previous studies on scorpion envenomation [4, 6, 13, 17, 23, 24, 72-74].

The histological alterations induced by Aah venom were accurately reflected in the serum by elevated CPK, ALT, ALP and LDH activities, which are commonly associated with the severity of tissue damage in cases of scorpion envenomation [6, 19]. These pathological manifestations are consistent with clinical observations in human scorpion envenomation. They include systemic inflammatory response, cardiovascular dysfunction, pulmonary edema [75-77] and hepatorenal manifestations [78-80]. Several studies have emphasized the relevance of pro-inflammatory mediators in the pathophysiological manifestations of human scorpion envenomation [16]. Moreover, there is a significant association between these complications and oxidative stress induced by venom components [81], as well as the release of organ dysfunction biomarkers such as LDH, CPK, and ALT, which are commonly elevated in clinical cases of envenomation and reflect systemic tissue injury [64, 78].The findings of the present study are in line with these clinical features, as experimental envenomation in mice led to increased vascular permeability, inflammatory infiltration, and organ damage. The observed increase in inflammatory and oxidative markers mirrors the pathophysiological processes seen in human victims, thereby validating the experimental model used.

The results of this study demonstrated that pretreatment of mice with a proteasome inhibitor introduced at low (0.05 mg/kg), medium (0.25 mg/kg) or high (0.5 mg/kg) doses prior to administration of the venom reduced the levels of inflammatory and oxidative markers seen in scorpion envenomation in a dose-dependent manner. A more preventive effect was observed at the cardiac, pulmonary and hepatic levels in the presence of a high concentration of bortezomib (0.5 mg/kg). In the renal tissue, it appears that the medium dose (0.25 mg/kg) of the proteasome inhibitor is more effective in reducing the immune inflammatory response. Several studies have demonstrated the effectiveness of these doses in reducing inflammation response during experimental inflammatory pathologies [82, 83]. In the present research, bortezomib appeared to prevent the increase in vascular permeability and the increase in tissue water content. These observations are supported by previous studies that link proteasome inhibition to reduced endothelial activation and permeability in inflammatory settings [84]. We did not directly assess the molecular markers of vascular leakage or tight junction integrity in the present study, but the reduction in Evans blue concentration in organs, tissue edema and MPO and EPO activities in treated animals indirectly supports a protective role of proteasome inhibition against vascular barrier dysfunction suggesting a stabilizing effect on endothelial integrity.

Although not directly assessed in this study, it is plausible that the proteasome inhibitor influenced cellular signaling networks such as NF-κB, which governs inflammatory responses. The NF-κB is a key driver of inflammation, its activation relies on proper proteasome function [85]. The NF-kB would be activated by the proteasome responsible for the degradation and cleavage of the IκB-NF-κB complex, the release of the factor NF-κB and its translocation into the nucleus for the expression of pro-inflammatory mediators [86].

The decrease in the rate of vascular permeability, which precedes the infiltration of inflammatory cells, may be due to the fact that the inhibition of the proteasome prior to envenomation can prevent the activation of the NF-kB factor, which could be activated by Toll-like receptors (TLR-2 and TLR-4) [73]. TLRs and their signaling pathways are currently being validated as potential immune-modulatory targets during scorpion envenomation pathogenesis [15, 17, 73]. Since these receptors are expressed in circulating and resident immune cells, including mast cells [87]. Inhibition of this signaling pathway prevents the release of preformed inflammatory mediators such as histamine and lipid derived mediators [6, 8], and thereby reduced vascular permeability. The decrease in peroxidase activities (MPO and EPO) in peripheral tissues after Aah envenomation in the presence of bortezomib could be partly attributed to the reduced vascular permeability observed in this study, which may have limited leukocyte extravasation. However, this reduction might also result from additional mechanisms, such as the apoptotic effects of proteasome inhibition on leukocytic cells, which may contribute to their immunosuppression [88-90]. It could also contribute to the reduction of the involved inflammatory mediators in the recruitment of polymorphonuclear cells such as cytokines (IL-2, IL-5, IL-8) [91] and adhesion molecules on the endothelium like E selectins, VCAM -1 and ICAM-1 and P-selectin [92-94].

The inhibition of proteasome resulted also in the prevention of oxidative balance alteration during Aah envenomation. This could be explained by the reduction in infiltrated neutrophil and eosinophil cells which represent important sources of reactive oxygen intermediates such as H2O2 and NO [66, 67]. Previous studies demonstrated also that increased production of ROS leads to the activation of NFĸB. This, in turn, stimulates the synthesis of other pro-inflammatory mediators such as adhesion molecules, which contribute to exacerbation of the inflammatory response [95]. It has been earlier shown that the inhibition of the proteasome system with the use of selective inhibitors blocks the induction of iNOS by preventing activation of nuclear factor‐κB [96-98]. This is associated with the upregulation of several endogenous antioxidants such as SOD, GSH‐Px and catalase contributing to significant improvement of tissue alteration [99]. This was in accordance with our results and with another previous report, which demonstrated that proteasome system inhibition may lead to increased antioxidant enzyme activities [100]. It reduces tissue oxidative damage which in turn prevents the release of the enzymatic cell contents (CPK, LDH, ALP, and ALT), as well as urea and creatinine levels.

The mechanism of the anti-inflammatory effect observed following inhibition of the proteasome prior to envenomation of mice might be more complex. Based on the results obtained in this study and supported by bibliographic data, we suggest that proteins modified by conjugation with lipid peroxidation products produced during scorpion envenomation may undergo ubiquitination and subsequent degradation by the proteasome [101, 102]. These degraded proteins can then be presented to T cells via a class 1 MHC molecule. The onset of a cytotoxic immune response can worsen the tissue damage observed after scorpion envenomation. According to these data, it is possible to suggest that during scorpion envenomation, tissue damage is mediated by both the Th2 and Th1 immune pathways involving the proteasome system [11].

Conclusion

In conclusion, the results of this study demonstrate, for the first time, the immunomodulatory role of the proteasome during scorpion envenomation. Administration of bortezomib effectively suppresses systemic inflammation by reducing vascular permeability, tissue edema, and peroxidase activities (MPO and EPO), while also enhancing the expression of antioxidants such as GSH and catalase. These effects ultimately contribute to the prevention of lipid peroxidation and tissue damage. Further investigations are warranted to explore the involvement of key inflammatory pathways, including NF-κB and Nrf2 signaling, as well as to elucidate the precise mechanisms underlying the proteasome-mediated regulation of inflammation and oxidative stress.

Abbreviations

Aah: Androctonus australis hector, ALP: alkaline phosphatase, ALT: alanine aminotransferase, ARE: antioxidant response element, CAT: catalase, COX-2: cyclooxygenase 2, CPK: creatine-phosphokinase, CRP: C-reactive protein, DTNB: 5,5'-dithiobis(2-nitrobenzoic acid), EDTA: ethylenediaminetetraacetic acid, EPO: eosinophil peroxidase, GPx: glutathione peroxidase, GSH: reduced glutathione, ICAM-1: intercellular adhesion molecule 1, iNOS: inducible nitric oxide synthase, LDH: lactate dehydrogenase, MDA: malondialdehyde, MHC: major histocompatibility complex, MPO: myeloperoxidase, NF-kB: nuclear factor kappa B, NMRI: Naval Medical Research Institute, NRf2: nuclear erythroid-2 like factor-2, OPD: o-phenylenediamine, PBS: phosphate-buffered saline, SDS: sodium dodecyl sulfate, SEM: standard error of the mean, TCA: trichloroacetic acid, TLRs: toll-like receptors, VCAM-1: vascular cell adhesion molecule 1.

Acknowledgments

The authors extend their heartfelt gratitude to all the participants who generously contributed their time and effort to this study.

Footnotes

Funding: This research was carried out without financial support from public agencies, commercial entities, or non-profit organizations.

Ethics approval and consent to participate: All animal procedures were conducted in compliance with the ethical standards set forth by the European Parliament and Council Directive on the protection of animals used for scientific purposes (Directive 2010/63/EU). This study was approved by the Deontology and Ethic Committee of the Research Thematic Agency in Health Science (ATRSS) formerly National Agency of Research Development in Health (ANDRS).

Consent for publication: Not applicable

Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

References

  1. Abroug F, Ouanes-Besbes L, Tilouche N, Elatrous S. Scorpion envenomation: state of the art. Intensive Care Med. 2020;46(3):401–410. doi: 10.1007/s00134-020-05924-8. [DOI] [PubMed] [Google Scholar]
  2. Boubekeur K, L’Hadj M, Selmane S. Demographic and epidemiological characteristics of scorpion envenomation and daily forecasting of scorpion sting counts in Touggourt, Algeria. Epidemiol health. 2020;42:e2020050. doi: 10.4178/epih.e2020050. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alhelail M, Albelali A, Alkanhal R, Salam M. Severity of scorpion envenomation in Saudi Arabia: A systematic review. Toxicol Rep. 2024:101749. doi: 10.1016/j.toxrep.2024.101749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bessalem S, Hammoudi-Triki D, Laraba-Djebari F. Effet de l’immunothérapie sur les modifications métaboliques et histopathologiques après envenimation scorpionique expérimentale. Bull Soc Pathol Exot. 2003;96(2):110–114. [PubMed] [Google Scholar]
  5. Heidarpour M, Ennaifer E, Ahari H, Srairi-Abid N, Borchani L, Khalili G, Amini H, Anvar AA, Boubaker S, El-Ayeb M, Shahbazzadeh D. Histopathological changes induced by Hemiscorpius lepturus scorpion venom in mice. Toxicon. 2012;59(3):373–378. doi: 10.1016/j.toxicon.2011.12.011. [DOI] [PubMed] [Google Scholar]
  6. Lamraoui A, Adi-Bessalem S, Laraba-Djebari F. Modulation of tissue inflammatory response by histamine receptors in scorpion envenomation pathogenesis: involvement of H4 receptor. Inflammation. 2014;37:1689–1704. doi: 10.1007/s10753-014-9898-x. [DOI] [PubMed] [Google Scholar]
  7. Chaïr-Yousfi I, Laraba-Djebari F, Hammoudi-Triki D. Androctonus australis hector venom contributes to the interaction between neuropeptides and mast cells in pulmonary hyperresponsiveness. Int Immunopharmacol. 2015;25(1):19–29. doi: 10.1016/j.intimp.2015.01.008. [DOI] [PubMed] [Google Scholar]
  8. Lamraoui A, Adi-Bessalem S, Laraba-Djebari F. Immunopathologic effects of scorpion venom on hepato-renal tissues: Involvement of lipid derived inflammatory mediators. Exp Mol Pathol. 2015;99(2):286–296. doi: 10.1016/j.yexmp.2015.07.013. [DOI] [PubMed] [Google Scholar]
  9. Haddad-Ishak-boushaki W, Laraba-Djebari F. Age-related changes in inflammatory response after experimental envenomation: impact on the susceptibility to Androctonus australis hector venom. Inflammation. 2017;40:1131–1142. doi: 10.1007/s10753-017-0557-x. [DOI] [PubMed] [Google Scholar]
  10. Ismail M. The scorpion envenoming syndrome. Toxicon. 1995;33(7):825–858. doi: 10.1016/0041-0101(95)00005-7. [DOI] [PubMed] [Google Scholar]
  11. Petricevich VL. Scorpion venom and the inflammatory response. Mediators Inflamm. 2010;2010(1):903295. doi: 10.1155/2010/903295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Magalhães MM, Pereira ME, Amaral CF, Rezende NA, Campolina D, Bucaretchi F, Gazzinelli RT, Cunha-Melo JR. Serum levels of cytokines in patients envenomed by Tityus serrulatus scorpion sting. Toxicon. 1999;37(8):1155–1164. doi: 10.1016/S0041-0101(98)00251-7. [DOI] [PubMed] [Google Scholar]
  13. Adi-Bessalem S, Mendil A, Hammoudi-Triki D, Laraba-Djebari F. Lung immunoreactivity and airway inflammation: their assessment after scorpion envenomation. Inflammation. 2012;35(2):501–508. doi: 10.1007/s10753-011-9338-0. [DOI] [PubMed] [Google Scholar]
  14. Ait-Lounis A, Laraba-Djebari F. TNF-alpha modulates adipose macrophage polarization to M1 phenotype in response to scorpion venom. Inflamm Res. 2015;64:929–936. doi: 10.1007/s00011-015-0876-z. [DOI] [PubMed] [Google Scholar]
  15. Zoccal KF, Paula-Silva FW, da Silva Bitencourt C, Sorgi CA, Bordon KD, Arantes EC, Faccioli LH. PPAR-γ activation by Tityus serrulatus venom regulates lipid body formation and lipid mediator production. Toxicon. 2015;93:90–97. doi: 10.1016/j.toxicon.2014.11.226. [DOI] [PubMed] [Google Scholar]
  16. Reis MB, Zoccal KF, Gardinassi LG, Faccioli LH. Scorpion envenomation and inflammation: Beyond neurotoxic effects. Toxicon. 2019;167:174–179. doi: 10.1016/j.toxicon.2019.06.219. [DOI] [PubMed] [Google Scholar]
  17. Khemili D, Laraba-Djebari F, Hammoudi-Triki D. Involvement of toll-like receptor 4 in neutrophil-mediated inflammation, oxidative stress and tissue damage induced by scorpion venom. Inflammation. 2020;43(1):155–167. doi: 10.1007/s10753-019-01105-y. [DOI] [PubMed] [Google Scholar]
  18. Fukuhara YD, Reis ML, Dellalibera-Joviliano R, Cunha FQ, Donadi EA. Increased plasma levels of IL-1β, IL-6, IL-8, IL-10 and TNF-α in patients moderately or severely envenomed by Tityus serrulatus scorpion sting. Toxicon. 2003;41(1):49–55. doi: 10.1016/S0041-0101(02)00208-8. [DOI] [PubMed] [Google Scholar]
  19. Hammoudi-Triki D, Ferquel E, Robbe-Vincent A, Bon C, Choumet V, Laraba-Djebari F. Epidemiological data, clinical admission gradation and biological quantification by ELISA of scorpion envenomations in Algeria: effect of immunotherapy. Transactions of the royal society of tropical medicine and hygiene. 2004;98(4):240–250. doi: 10.1016/S0035-9203(03)00062-2. [DOI] [PubMed] [Google Scholar]
  20. Bekkari N, Martin-Eauclaire MF, Laraba-Djebari F. Complement system and immunological mediators: their involvements in the induced inflammatory process by Androctonus australis hector venom and its toxic components. Exp Toxicol Pathol. 2015;67(7-8):389–397. doi: 10.1016/j.etp.2015.04.002. [DOI] [PubMed] [Google Scholar]
  21. Medjadba W, Martin-Eauclaire MF, Laraba-Djebari F. Involvement of kallikrein-Kinin system on cardiopulmonary alterations and inflammatory response induced by purified Aah I toxin from scorpion venom. Inflammation. 2016;39:290–302. doi: 10.1007/s10753-015-0249-3. [DOI] [PubMed] [Google Scholar]
  22. Nakib I, Martin-Eauclaire MF, Laraba-Djebari F. Involvement of cholinergic and adrenergic receptors in pathogenesis and inflammatory response induced by alpha-neurotoxin Bot III of scorpion venom. Inflammation. 2016;39:1670–1680. doi: 10.1007/s10753-016-0401-8. [DOI] [PubMed] [Google Scholar]
  23. Sifi A, Adi-Bessalem S, Laraba-Djebari F. Role of angiotensin II and angiotensin type-1 receptor in scorpion venom-induced cardiac and aortic tissue inflammation. Exp Mol Pathol. 2017;102(1):32–40. doi: 10.1016/j.yexmp.2016.11.006. [DOI] [PubMed] [Google Scholar]
  24. Sifi A, Adi-Bessalem S, Laraba-Djebari F. Involvement of the endothelin receptor type A in the cardiovascular inflammatory response following scorpion envenomation. Toxins. 2020;12(6):389–389. doi: 10.3390/toxins12060389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Dousset E, Carrega L, Steinberg JG, Clot-Faybesse O, Jouirou B, Sauze N, Devaux C, Autier Y, Jammes Y, Martin-Eauclaire MF, Guieu R. Evidence that free radical generation occurs during scorpion envenomation. Comp Biochem Physiol C. 2005;140(2):221–226. doi: 10.1016/j.cca.2005.02.003. [DOI] [PubMed] [Google Scholar]
  26. Raouraoua-Boukari R, Sami-Merah S, Hammoudi-Triki D, Martin-Eauclaire MF, Laraba-Djebari F. Immunomodulation of the inflammatory response induced by Androctonus australis hector neurotoxins: biomarker interactions. Neuroimmunomodulation. 2012;19(2):103–110. doi: 10.1159/000330241. [DOI] [PubMed] [Google Scholar]
  27. Galvani NC, Vilela TC, Domingos AC, Fagundes MÍ, Bosa LM, Della Vechia IC, Scussel R, Pereira M, Steiner BT, Damiani AP, Chávez-Olórtegui C. Genotoxicity evaluation induced by Tityus serrulatus scorpion venom in mice. Toxicon. 2017;140:132–138. doi: 10.1016/j.toxicon.2017.10.024. [DOI] [PubMed] [Google Scholar]
  28. Khadem MD, Rasooli A, Mashhadi AG, Shahriari A, Mohammadian B, Barati F. Evaluation of oxidant/antioxidant status in serum of sheep experimentally envenomated with Hemiscorpius lepturus scorpion venom. Vet Res Forum. 2019;10(2):165. doi: 10.30466/vrf.2018.80273.2066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Daachi F, Adi-Bessalem S, Megdad-Lamraoui A, Laraba-Djebari F. Immune-toxicity effects of scorpion venom on the hypothalamic pituitary adrenal axis during rest and activity phases in a rodent model. Comp Biochem Physiol C. 2020;235:108787. doi: 10.1016/j.cbpc.2020.108787. [DOI] [PubMed] [Google Scholar]
  30. Chen C, Qin H, Tan J, Hu Z, Zeng L. The role of ubiquitin-proteasome pathway and autophagy-lysosome pathway in cerebral ischemia. Oxid Med Cell Longev. 2020;2020(1):5457049. doi: 10.1155/2020/5457049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Korolchuk VI, Menzies FM, Rubinsztein DC. Mechanisms of cross-talk between the ubiquitin-proteasome and autophagy-lysosome systems. FEBS Letters. 2010;584(7):1393–1398. doi: 10.1016/j.febslet.2009.12.047. [DOI] [PubMed] [Google Scholar]
  32. Adams J. The proteasome: structure, function, and role in the cell. Cancer Treat Rev. 2003;29:3–9. doi: 10.1016/S0305-7372(03)00081-1. [DOI] [PubMed] [Google Scholar]
  33. Ciechanover A. Intracellular protein degradation: from a vague idea, through the lysosome and the ubiquitin-proteasome system, and onto human diseases and drug targeting (Nobel lecture) Angew Chem Int Ed Engl. 2005;44(37):5944–5967. doi: 10.1002/anie.200501428. [DOI] [PubMed] [Google Scholar]
  34. Loureiro J, Ploegh HL. Antigen presentation and the ubiquitin-proteasome system in host-pathogen interactions. Adv Immunol. 2006;92:225–305. doi: 10.1016/S0065-2776(06)92006-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Traenckner EB, Wilk S, Baeuerle PA. A proteasome inhibitor prevents activation of NF-kappa B and stabilizes a newly phosphorylated form of I kappa B-alpha that is still bound to NF-kappa B. EMBO J. 1994;13(22):5433–5441. doi: 10.1002/j.1460-2075.1994.tb06878.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Chen ZJ, Maniatis T. Role of the Ubiquitin-Proteasome Pathway in NF-κB Activation. InUbiquitin and the Biology of the Cell. Boston, MA: Springer US; 1998. pp. 303–322. [DOI] [Google Scholar]
  37. Monsalvo-Maraver LA, Ovalle-Noguez EA, Nava-Osorio J, Maya-López M, Rangel-López E, Túnez I, Tinkov AA, Tizabi Y, Aschner M, Santamaría A. Interactions Between the Ubiquitin-Proteasome System, Nrf2, and the Cannabinoidome as Protective Strategies to Combat Neurodegeneration: Review on Experimental Evidence. Neurotox Res. 2024;42(18) doi: 10.1007/s12640-024-00694-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Laraba-Djebari F, Hammoudi D. Use of toxic fraction isolated from Algerian Androctonus australis hector scorpion venom for the assessment of anti-venom serum. Arch Inst Pasteur Alger. 1998;62:254–266. [PubMed] [Google Scholar]
  39. Adi-Bessalem S, Hammoudi-Triki D, Laraba-Djebari F. Pathophysiological effects of Androctonus australis hector scorpion venom: tissue damages and inflammatory response. Experimental and toxicologic pathology. 2008;60(4-5):373–380. doi: 10.1016/j.etp.2008.03.006. [DOI] [PubMed] [Google Scholar]
  40. Matos IM, Souza DG, Seabra DG, Freire-Maia L, Teixeira MM. Effects of tachykinin NK1 or PAF receptor blockade on the lung injury induced by scorpion venom in rats. Eur J Pharmacol. 1999;376(3):293–300. doi: 10.1016/S0014-2999(99)00382-9. [DOI] [PubMed] [Google Scholar]
  41. Coelho FM, Pessini AC, Coelho AM, Pinho VS, Souza DG, Arantes EC, Teixeira MM, Teixeira AL. Platelet activating factor receptors drive CXC chemokine production, neutrophil influx and edema formation in the lungs of mice injected with Tityus serrulatus venom. Toxicon. 2007;50(3):420–427. doi: 10.1016/j.toxicon.2007.04.009. [DOI] [PubMed] [Google Scholar]
  42. Van Oosterhout AJ, Van Ark I, Hofman G, Van Der Linde HJ, Fattah D, Nijkamp FP. Role of interleukin-5 and substance P in development of airway hyperreactivity to histamine in guinea-pigs. Eur Respir J. 1996;9(3):493–499. doi: 10.1183/09031936.96.09030493. [DOI] [PubMed] [Google Scholar]
  43. Loscalzo J. What we know and don’t know about L-arginine and NO. Circulation. 2000;101(18):2126–2129. doi: 10.1161/01.CIR.101.18.2126. [DOI] [PubMed] [Google Scholar]
  44. Pick E, Keisari Y. A simple colorimetric method for the measurement of hydrogen peroxide produced by cells in culture. J Immunol Methods. 1980;38(1-2):161–170. doi: 10.1016/0022-1759(80)90340-3. [DOI] [PubMed] [Google Scholar]
  45. Moreno I, Pichardo S, Jos A, Gomez-Amores L, Mate A, Vazquez CM, Camean AM. Antioxidant enzyme activity and lipid peroxidation in liver and kidney of rats exposed to microcystin-LR administered intraperitoneally. Toxicon. 2005;45(4):395–402. doi: 10.1016/j.toxicon.2004.11.001. [DOI] [PubMed] [Google Scholar]
  46. Aebi H. In: Methods of enzymatic analysis. Bergmeyer HU, editor. Weinheim: NewYork: Verlag Chemie; Academic Press Inc; 1974. Catalase; pp. 673–684. [DOI] [Google Scholar]
  47. Zhang Y. Role of glutathione in the accumulation of anticarcinogenic isothiocyanates and their glutathione conjugates by murine hepatoma cells. Carcinogenesis. 2000;21(6):1175–1182. doi: 10.1093/carcin/21.6.1175. [DOI] [PubMed] [Google Scholar]
  48. Meki AR, El-Dean ZM. Serum interleukin-1β, interleukin-6, nitric oxide and α1-antitrypsin in scorpion envenomed children. Toxicon. 1998;36(12):1851–1859. doi: 10.1016/S0041-0101(98)00106-8. [DOI] [PubMed] [Google Scholar]
  49. Borges CM, Silveira MR, Aparecida M, Beker CL, Freire-Maia L, Teixeira MM. Scorpion venom-induced neutrophilia is inhibited by a PAF receptor antagonist in the rat. J Leukoc Biol. 2000;67(4):515–519. doi: 10.1016/S0041-0101(98)00106-8. [DOI] [PubMed] [Google Scholar]
  50. Pessini AC, de Souza AM, Faccioli LH, Gregório ZM, Arantes EC. Time course of acute-phase response induced by Tityus serrulatus venom and TsTX-I in mice. Int Immunopharmacol. 2003;3(5):765–774. doi: 10.1016/S1567-5769(03)00078-X. [DOI] [PubMed] [Google Scholar]
  51. Kaddache A, Hassan M, Laraba-Djebari F, Hammoudi-Triki D. Switch of steady-state to an accelerated granulopoiesis in response to Androctonus australis hector venom. Inflammation. 2017;40:871–883. doi: 10.1007/s10753-017-0532-6. [DOI] [PubMed] [Google Scholar]
  52. Siddall EC, Radhakrishnan J. The pathophysiology of edema formation in the nephrotic syndrome. Kidney Int. 2012;82(6):635–642. doi: 10.1038/ki.2012.180. [DOI] [PubMed] [Google Scholar]
  53. Zuliani JP, Freitas TA, Conceição IM, Kwasniewski FH. Tityus serrulatus venom increases vascular permeability in selected airway tissues in a mast cell-independent way. Exp Toxicol Pathol. 2013;65(3):229–234. doi: 10.1016/j.etp.2011.08.010. [DOI] [PubMed] [Google Scholar]
  54. Fatani AJ, Al-Zuhair HA, Yaquob HI, Abdel-Fattah AA, El-Sayed MI, El-Sayed FA. Protective effects of the antioxidant Ginkgo biloba extract and the protease inhibitor aprotinin against Leiurus quinquestriatus venom-induced tissue damage in rats. J Venom Anim Toxinsincl Trop Dis. 2006;12:255–275. doi: 10.1590/S1678-91992006000200008. [DOI] [Google Scholar]
  55. Santos AC, Lopes C, Guimaraes JT, Barros H. Central obesity as a major determinant of increased high-sensitivity C-reactive protein in metabolic syndrome. Int J Obes. 2005;29(12):1452–1456. doi: 10.1038/sj.ijo.0803035. [DOI] [PubMed] [Google Scholar]
  56. Tohamy AA, Mohamed AF, Moneim AEA, Diab MS. Biological effects of Naja haje crude venom on the hepatic and renal tissues of mice. J King Saud Univ Sci. 2014;26:205–212. [Google Scholar]
  57. Hsu SM, Yang CH, Shen FH, Chen SH, Lin CJ, Shieh CC. Proteasome Inhibitor Bortezomib Suppresses Nuclear Factor-Kappa B Activation and Ameliorates Eye Inflammation in Experimental Autoimmune Uveitis. Mediators Inflamm. 2015;2015(1):847373. doi: 10.1155/2015/847373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Chen FT, Liu YC, Yang CM, Yang CH. Anti-inflammatory effect of the proteasome inhibitor bortezomib on endotoxin-induced uveitis in rats. Invest Ophthalmol Vis Sci. 2012;53(7):3682–3694. doi: 10.1167/iovs.12-9505. [DOI] [PubMed] [Google Scholar]
  59. Campos F, Abrigo J, Aguirre F, Garcés B, Arrese M, Karpen S, Cabrera D, Andía ME, Simon F, Cabello-Verrugio C. Sarcopenia in a mice model of chronic liver disease: role of the ubiquitin-proteasome system and oxidative stress. Pflügers Archiv-Eur J Physiol. 2018;470:1503–1519. doi: 10.1007/s00424-018-2167-3. [DOI] [PubMed] [Google Scholar]
  60. Goetzke CC, Ebstein F, Kallinich T. Role of proteasomes in inflammation. J Clin Med. 2021;10(8):1783. doi: 10.3390/jcm10081783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Ruscitti P, Currado D, Rivellese F, Vomero M, Navarini L, Cipriani P, Pitzalis C, Giacomelli R. Diminished expression of the ubiquitin-proteasome system in early treatment-naïve patients with rheumatoid arthritis and concomitant type 2 diabetes may be linked to IL-1 pathway hyper-activity; results from PEAC cohort. Arthritis Res Ther. 2024;26(1):171. doi: 10.1186/s13075-024-03392-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  62. Megdad-Lamraoui A, Adi-Bessalem S, Laraba-Djebari F. Cerebrospinal inflammatory response following scorpion envenomation: role of histamine H1 and H3 receptors. Inflammopharmacology. 2019;27:589–601. doi: 10.1007/s10787-018-00553-6. [DOI] [PubMed] [Google Scholar]
  63. Sami-Merah S, Hammoudi-Triki D, Martin-Eauclaire MF, Laraba-Djebari F. Combination of two antibody fragments F (ab′) 2/Fab: An alternative for scorpion envenoming treatment. Int Immunopharmacol. 2008;8(10):1386–1394. doi: 10.1016/j.intimp.2008.05.011. [DOI] [PubMed] [Google Scholar]
  64. Cupo P, Hering SE. Cardiac troponin I release after severe scorpion envenoming by Tityus serrulatus. Toxicon. 2002;40(6):823–830. doi: 10.1016/S0041-0101(02)00080-6. [DOI] [PubMed] [Google Scholar]
  65. Soeters PB, Wolfe RR, Shenkin A. Hypoalbuminemia: pathogenesis and clinical significance. J Parenter Enteral Nutr. 2019;43(2):181–193. doi: 10.1002/jpen.1451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  66. Elsner J, Dichmann S, Kapp A. Activation of the respiratory burst in human eosinophils by chemotaxins requires intracellular calcium fluxes. J Invest Dermatol. 1995;105:231–236. doi: 10.1111/1523-1747.ep12317519. [DOI] [PubMed] [Google Scholar]
  67. Defraigne JO, Pincemail J, Larbuisson R, Blaffart F, Limet R. Cytokine release and neutrophil activation are not prevented by heparin-coated circuits and aprotinin administration. Ann Thorac Surg. 2000;69(4):1084–1091. doi: 10.1016/S0003-4975(00)01093-6. [DOI] [PubMed] [Google Scholar]
  68. Kimura H, Sawada T, Oshima S, Kozawa K, Ishioka T, Kato M. Toxicity and roles of reactive oxygen species. Curr Drug Targets Inflamm Allergy. 2005;4(4):489–495. doi: 10.2174/1568010054526287. [DOI] [PubMed] [Google Scholar]
  69. Lardinois OM, Mestdagh MM, Rouxhet PG. Reversible inhibition and irreversible inactivation of catalase in presence of hydrogen peroxide. Biochim Biophys Acta. 1996;1295(2):222–238. doi: 10.1016/0167-4838(96)00043. [DOI] [PubMed] [Google Scholar]
  70. Folkerts G, Kloek J, Muijsers RB, Nijkamp FP. Reactive nitrogen and oxygen species in airway inflammation. Eur J Pharmacol. 2001;429(1-3):251–262. doi: 10.1016/S0014-2999(01)01324-3. [DOI] [PubMed] [Google Scholar]
  71. Daachi F, Adi-Bessalem S, Megdad-Lamraoui A, Laraba-Djebari F. Influence of envenomation timing on peripheral immune and oxidative responses in experimental scorpion envenomation. J Venom Anim Toxinsincl Trop Dis. 2025;31:e20240059. doi: 10.1590/1678-9199-JVATITD-2024-0059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  72. Boussag-Abib L, Laraba-Djebari F. InImmunology. Vol. 135. MA USA: WILEY-BLACKWELL; 2011. Safe antigens: their use in immune sera production and vaccine approach; pp. 204–204. COMMERCE PLACE, 350 MAIN ST, MALDEN 02148. [DOI] [Google Scholar]
  73. Zoccal KF, Bitencourt CD, Paula-Silva FW, Sorgi CA, de Castro Figueiredo Bordon K, Arantes EC, Faccioli LH. TLR2, TLR4 and CD14 recognize venom-associated molecular patterns from Tityus serrulatus to induce macrophage-derived inflammatory mediators. PLoS One. 2014;9(2):e88174. doi: 10.1371/journal.pone.0088174. [DOI] [PMC free article] [PubMed] [Google Scholar]
  74. Taibi-Djennah Z, Laraba-Djebari F. Effect of cytokine antibodies in the immunomodulation of inflammatory response and metabolic disorders induced by scorpion venom. Int Immunopharmacol. 2015;27(1):122–129. doi: 10.1016/j.intimp.2015.05.002. [DOI] [PubMed] [Google Scholar]
  75. Bahloul M, Chabchoub I, Chaari A, Chtara K, Kallel H, Dammak H, Ksibi H, Chelly H, Rekik N, Hamida CB, Bouaziz M. Scorpion envenomation among children: clinical manifestations and outcome (analysis of 685 cases) Am J Trop Med Hyg. 2010;83(5):1084. doi: 10.4269/ajtmh.2010.10-0036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  76. Bahloul M, Chaari A, Dammak H, Samet M, Chtara K, Chelly H, Hamida CB, Kallel H, Bouaziz M. Pulmonary edema following scorpion envenomation: mechanisms, clinical manifestations, diagnosis and treatment. Int J Cardiol. 2013;162(2):86–91. doi: 10.1016/j.ijcard.2011.10.013. [DOI] [PubMed] [Google Scholar]
  77. Abroug F, Souheil E, Ouanes I, Dachraoui F, Fekih-Hassen M, Ouanes Besbes L. Scorpion-related cardiomyopathy: clinical characteristics, pathophysiology, and treatment. Clin Toxicol. 2015;53(6):511–518. doi: 10.3109/15563650.2015.1030676. [DOI] [PubMed] [Google Scholar]
  78. Valavi E, Ansari MA. Hemolytic uremic syndrome following Hemiscorpius lepturus (scorpion) sting. Indian J Nephrol. 2008;18(4):166–168. doi: 10.4103/0971-4065.45293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  79. Bahloul M, Hmida B, Belhoul W, Ksibi H, Kallel H, Chaari A, Chelly H, Rekik N, Bouaziz M. Hemolytic-uremic syndrome secondary to scorpion envenomation (apropos of 2 cases) Nephrologie. 2004;25(2):49–51. [PubMed] [Google Scholar]
  80. Krkic-Dautovic S, Begovic B. Acute renal insufficiency and toxic hepatitis following scorpions sting. Medicinski Arhiv. 2007;61(2):123–124. [PubMed] [Google Scholar]
  81. Al B, Yarbil P, Zengin S, Ercan S, Davutoglu V, Yildirim C. Impact of scorpion stings on electrocardiographic changes and relationship with body oxidant and antioxidant status. J Pak Med Assoc. 2014;64:423–427. [PubMed] [Google Scholar]
  82. Mao X, Pan X, Cheng T, Zhang X. Therapeutic potential of the proteasome inhibitor Bortezomib on titanium particle-induced inflammation in a murine model. Inflammation. 2012;35:905–912. doi: 10.1007/s10753-011-9392-7. [DOI] [PubMed] [Google Scholar]
  83. Wagner-Ballon O, Pisani DF, Gastinne T, Tulliez M, Chaligné R, Lacout C, Auradé F, Villeval JL, Gonin P, Vainchenker W, Giraudier S. Proteasome inhibitor bortezomib impairs both myelofibrosis and osteosclerosis induced by high thrombopoietin levels in mice. Blood. 2007;110(1):345–353. doi: 10.1182/blood-2006-10-054502. [DOI] [PubMed] [Google Scholar]
  84. Lorenz M, Wilck N, Meiners S, Ludwig A, Baumann G, Stangl K, Stangl V. Proteasome inhibition prevents experimentally-induced endothelial dysfunction. Life Sci. 2009;84(25-26):929–934. doi: 10.1016/j.lfs.2009.04.016. [DOI] [PubMed] [Google Scholar]
  85. Yin L, Laevsky G, Giardina C. Butyrate suppression of colonocyte NF-κB activation and cellular proteasome activity. J Biol Chem. 2001;276(48):44641–44646. doi: 10.1074/jbc.M105170200. [DOI] [PubMed] [Google Scholar]
  86. Amit S, Ben-Neriah Y. NF-κB activation in cancer: a challenge for ubiquitination-and proteasome-based therapeutic approach. Semin Cancer Biol. 2003;13(1):15–28. doi: 10.1016/S1044-579X(02)00096-2. [DOI] [PubMed] [Google Scholar]
  87. Smrž D, Iwaki S, McVicar DW, Metcalfe DD, Gilfillan AM. TLR-mediated signaling pathways circumvent the requirement for DAP12 in mast cells for the induction of inflammatory mediator release. Eur J Immunol. 2010;40(12):3557–3569. doi: 10.1002/eji.201040573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  88. Moran E, Carbone F, Augusti V, Patrone F, Ballestrero A, Nencioni A. Proteasome inhibitors as immunosuppressants: biological rationale and clinical experience. Semin Hematol. 2012;49(3):270–276. doi: 10.1053/j.seminhematol.2012.04.004. [DOI] [PubMed] [Google Scholar]
  89. Pellom ST, Jr, Dudimah DF, Thounaojam MC, Sayers TJ, Shanker A. Modulatory effects of bortezomib on host immune cell functions. Immunotherapy. 2015;7(9):1011–1022. doi: 10.2217/imt.15.66. [DOI] [PMC free article] [PubMed] [Google Scholar]
  90. Mohty M, Brissot E, Savani BN, Gaugler B. Effects of bortezomib on the immune system: a focus on immune regulation. Biol Blood Marrow Transplant. 2013;19(10):1416–1420. doi: 10.1016/j.bbmt.2013.05.011. [DOI] [PubMed] [Google Scholar]
  91. Guilpain P, Guillevin L, Mouthon L. Eosinophil granule cationic proteins: eosinophil activation markers. 2006. [DOI] [PubMed] [Google Scholar]
  92. Astarci E, Sade A, Cimen I, Savaş B, Banerjee S. The NF-κB target genes ICAM-1 and VCAM-1 are differentially regulated during spontaneous differentiation of Caco-2 cells. FEBS J. 2012;279(16):2966–2986. doi: 10.1111/j.1742-4658.2012.08677.x. [DOI] [PubMed] [Google Scholar]
  93. Xia YF, Liu LP, Zhong CP, Geng JG. NF-κB activation for constitutive expression of VCAM-1 and ICAM-1 on B lymphocytes and plasma cells. Biochem Biophys Res Commun. 2001;289(4):851–856. doi: 10.1006/bbrc.2001.6067. [DOI] [PubMed] [Google Scholar]
  94. Mocanu CA, Fuior EV, Voicu G, Rebleanu D, Safciuc F, Deleanu M, Fenyo IM, Escriou V, Manduteanu I, Simionescu M, Calin M. P-selectin targeted RAGE-shRNA lipoplexes alleviate atherosclerosis-associated inflammation. J Control Release. 2021;338:754–772. doi: 10.1016/j.jconrel.2021.09.012. [DOI] [PubMed] [Google Scholar]
  95. Gloire G, Legrand-Poels S, Piette J. NF-κB activation by reactive oxygen species: fifteen years later. Biochem Pharmacol. 2006;72(11):1493–1505. doi: 10.1016/j.bcp.2006.04.011. [DOI] [PubMed] [Google Scholar]
  96. Lee M, Hyun DH, Jenner P, Halliwell B. Effect of proteasome inhibition on cellular oxidative damage, antioxidant defences and nitric oxide production. J Neurochem. 2001;78(1):32–41. doi: 10.1046/j.1471-4159.2001.00416.x. [DOI] [PubMed] [Google Scholar]
  97. Griscavage JM, Wilk S, Ignarro LJ. Inhibitors of the proteasome pathway interfere with induction of nitric oxide synthase in macrophages by blocking activation of transcription factor NF-kappa B. Proc Nat Acad Sci. 1996;93(8):3308–3312. doi: 10.1073/pnas.93.8.3308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  98. Conner EM, Brand S, Davis JM, Laroux FS, Palombella VJ, Fuseler JW, Kang DY, Wolf RE, Grisham MB. Proteasome inhibition attenuates nitric oxide synthase expression, VCAM-1 transcription and the development of chronic colitis. J Pharmacol Exp Ther. 1997;282(3):1615–1622. [PubMed] [Google Scholar]
  99. Hu LH, Fan YJ, Li Q, Guan JM, Qu B, Pei FH, Liu BR. Bortezomib protects against dextran sulfate sodium-induced ulcerative colitis in mice. Mol Med Rep. 2017;15(6):4093–4099. doi: 10.3892/mmr.2017.6524. [DOI] [PMC free article] [PubMed] [Google Scholar]
  100. Alexandrova A, Petrov L, Georgieva A, Kirkova M, Kukan M. Effects of proteasome inhibitor, MG132, on proteasome activity and oxidative status of rat liver. Cell Biochem Funct. 2008;26(3):392–398. doi: 10.1111/j.1872-034X.2007.00270.x. [DOI] [PubMed] [Google Scholar]
  101. Jung T, Grune T. The proteasome and its role in the degradation of oxidized proteins. IUBMB Life. 2008;60(11):743–752. doi: 10.1002/iub.114. [DOI] [PubMed] [Google Scholar]
  102. Kandel R, Jung J, Neal S. Proteotoxic stress and the ubiquitin proteasome system. Semin Cell Dev Biol. 2024;156:107–120. doi: 10.1016/j.semcdb.2023.08.002. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Data Availability Statement

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Articles from The Journal of Venomous Animals and Toxins Including Tropical Diseases are provided here courtesy of Centro de Estudos de Venenos e Animais Peçonhentos - CEVAP

RESOURCES