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. 2025 Mar 8;104(4):104990. doi: 10.1016/j.psj.2025.104990

Dietary selenium mitigates cadmium-induced apoptosis and inflammation in chicken testicles by inhibiting oxidative stress through the activation of the Nrf2/HO-1 signaling pathway

Yulong Li a,c,⁎, Shu Wang b, Rui Feng c
PMCID: PMC11951179  PMID: 40081173

Abstract

Cadmium (Cd) is a non-essential heavy metal that is highly toxic to testicle. Selenium (Se) is known to possess antagonistic effects against Cd toxicity, yet the precise mechanisms through which Se counteracts Cd-induced testicular damage in chickens through Nuclear factor erythroid 2-related factor 2/Heme oxygenase-1 (Nrf2/HO-1) signaling pathway, oxidative stress (OS), apoptosis, and inflammation remained unclear. In the present study, the experimental model of chicken testis was established by incorporating CdCl2 and Na2SeO3 into the dietary intake. After 60 days, chickens from each group were euthanized, and testicular and serum samples were subsequently collected. Ultrastructural assessment revealed that Se supplementation significantly mitigated the testicular damage induced by Cd. Se effectively suppressed the Cd-induced elevation in ROS, MDA, and H2O2 levels, while also preventing the downregulation of CAT, GSH, and T-AOC levels. Furthermore, Se administration ameliorated the reduction in the expression levels of Nrf2, HO-1, and Bcl-2 induced by Cd, and counteracted the overexpression of Caspase-3, Bax, Cyt-c, and Caspase-9, TNF-α, IL-2, IL-6, and IL-1β. Meanwhile, immunofluorescence data demonstrated that Se attenuated the Cd-induced decrease in Nrf2 and HO-1 expression and the upregulation of IL-6 expression. In conclusion, this study elucidated that Se might mitigate Cd-induced oxidative stress in chicken testicles through the stimulation of the Nrf2/HO-1 signaling pathway, thereby inhibiting apoptosis and inflammation, and was beneficial in reducing Cd-induced testicular injury.

Keywords: Cadmium, Selenium, Oxidative stress, Apoptosis, Inflammation

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Introduction

Many regions worldwide are facing the issue of environmental and ecological destruction caused by pollutants such as chemical pesticides and heavy metals. These pollutants engage in material and energy exchanges with plants, animals, and humans through the biological chain, which can inflict damage upon biological organisms. Cadmium (Cd) is a heavy metal environmental pollutant distinguished by its long half-life and resistance to degradation (Bautista et al., 2024). As a result, Cd persists in the atmosphere, hydrosphere, and biosphere, entering the bodies of humans and animals through air, water, and food, leading to irreversible damage to various organs (Suhani et al., 2021; Li et al., 2024a). Currently, there is evidence indicating that exposure to Cd can lead to detrimental effects on several organs, including the liver and kidney (Fan et al., 2018). Furthermore, testicle is identified as a primary target for Cd-induced damage (Zhang et al., 2024a). Research has demonstrated that Cd accumulation is linked to a reduction in sperm count as well as a suppression of genes that contribute to the production of testosterone (Li et al., 2018). Research indicates that Cd inflicts damage to the testis through multiple mechanisms, including apoptosis, autophagy, and inflammatory responses. A related study reports that the TLR4/MAPK/NF-κB signaling pathway is implicated in Cd-induced apoptosis and necroptosis in pig testicular cells (Li et al., 2022). Additionally, related research has revealed that Cd can trigger the NLRP3/Caspase-1/IL-1β signaling pathway in rat testes, precipitating inflammation and apoptosis (Fouad et al., 2019).

Oxidative stress represents an imbalance between the antioxidant and pro-oxidant systems in cells, with the pro-oxidant system predominating (Hou et al., 2024). Antioxidant enzymes, including SOD, GSH-PX, and CAT, collectively constitute an antioxidant defense system that counteracts excessive ROS (Liang et al., 2024). Research has found that oxidative stress can modulate cellular injury processes such as apoptosis, inflammation, and necroptosis. Studies have shown that oxidative stress triggers apoptosis in renal tubular epithelial cells, resulting in cellular harm (Zhuang et al., 2019). Additionally, oxidative stress can lead to both inflammation and necroptosis in the small intestine of pigs (Chen et al., 2022). Nrf2, a member of the CNC family of transcription factors, plays a crucial role in the defense against oxidative damage and is ubiquitously expressed across various organs and tissues (Valipour et al., 2024). Heme oxygenase-1 (HO-1), a significant downstream molecule of Nrf2, possesses anti-inflammatory, antioxidant, and anti-apoptotic properties (Binmahfouz et al., 2024). Notably, under conditions of oxidative stress, Nrf2 promotes HO-1 upregulation, enhancing the body's antioxidant capacity and elevating antioxidant defenses. Consequently, the Nrf2/HO-1 axis forms a key mechanism for combating oxidative stress. Studies have indicated that Nrf2 and HO-1 are found in the heart and jejunum, and these molecules are deeply engaged in the modulation of diverse physiological functions, including oxidative stress, cell death, and inflammation. Evidence suggests that deltamethrin mitigates oxidative stress-induced apoptosis and inflammation in cardiomyocytes by the stimulation of the Nrf2/HO-1 signaling pathway (Yang et al., 2022). Furthermore, scientists have also identified that SeMet inhibits oxidative stress in the jejunum by stimulating the Nrf2/HO-1 signaling pathway, lowering oxidative harm and inflammatory responses (Zhang et al., 2022).

Selenium (Se) is an essential nutrient with a myriad of beneficial effects, including the enhancement of immune function, anti-inflammatory action, and cancer prevention (Ibrahim et al., 2024; Hughes et al., 2023). Therefore, Se is intimately connected with the health of humans and mammals. Se as an ideal detoxifying agent capable of countering the toxic effects induced by Cd (Wang et al., 2024). Studies have demonstrated that Se can suppress Cd-induced oxidative stress and apoptosis in chicken ovaries (Wan et al., 2017). Furthermore, research reports that Se mitigates the inflammatory responses through the NF-kB/IκB signaling pathway caused in chicken heart by Cd (Ge et al., 2021). However, research on the antagonistic effect of Se on Cd-mediated apoptosis and inflammatory responses in chicken testicles, particularly through the Nrf2/HO-1 signaling pathway and oxidative stress, remained unclear. Thus, the present study established a co-treatment model with Cd and Se to observe the situation of testicular injury in chickens. Subsequently, the Nrf2/HO-1 signaling pathway markers, oxidative stress indicators, and markers for both apoptosis and inflammation were carefully analyzed. This research may offer a theoretical foundation for further investigation into the mechanisms by which Se ameliorates the male reproductive toxicity of Cd and provide novel insights for the field of comparative medicine.

Materials and methods

Chemicals

Cadmium chloride (CdCl2) (purity ≥ 98 %) was sourced from Tianjin Zhiyuan Chemical Reagent Co., Ltd., Tianjin, China. Sodium selenite (Na2SeO3) (purity ≥ 98 %) was procured from Aladdin Biochemical Technology Co., Ltd., Shanghai, China.

Animals and experimental design

A total of 150 seven-day-old male chickens were randomly assigned to six groups, with 25 chickens per group (n = 25/group). The control group (Con) was provided with a basal diet. The diet of the low-selenium group (LS) was augmented with a 0.6 mg/kg supplement of Na2SeO3, while the high-selenium group (HS) was given a diet containing 1.0 mg/kg of Na2SeO3. The Cadmium group (Cd) was fed a basal diet with an addition of 150.00 mg/kg of CdCl2 (Wang et al., 2023b). The Cadmium plus low-selenium group (Cd+LS) was administered a diet containing both 150.00 mg/kg of CdCl2 and 0.6 mg/kg of Na2SeO3, and the Cadmium plus high-selenium group (Cd+HS) was fed a diet with 150.00 mg/kg of CdCl2 and 1.0 mg/kg of Na2SeO3. All chickens were fed in the same house environment, ensuring regular ventilation and continuous access to clean drinking water and adequate feed supply. Environmental conditions, including temperature, humidity, and light cycle, were all regulated to meet the welfare requirements. The chickens were raised under identical environmental conditions for 2 months. On day 60, all experimental chickens were fasted 24 h before sacrifice. Subsequently, testicular and blood samples were collected for analysis.

Ultrastructural observation

Select testicular tissue samples that were smaller than 1 cubic millimeter and fix them using a solution of 2.5 % glutaraldehyde. Then, the samples were subjected to a dehydration process, which involved incrementally raising the concentrations of alcohol. We used resin as the embedding medium to embed the dehydrated tissue. Next, the embedded tissue blocks were cured at different temperatures. An ultramicrotome was used to cut the solidified blocks into 70 nm thick sections. Finally, uranyl acetate and lead citrate were used to stain sections. For observation, a transmission electron microscope (TEM) (H-7650, Hitachi, Japan) was used to visualize and capture images of the sample.

Testosterone content analysis

After thawing the frozen blood samples at room temperature, we centrifuged them at 2000 rpm for 20 min to obtain the serum for subsequent analysis. The testosterone content was determined using a commercial assay kit provided by the Nanjing Jiancheng Bioengineering Institute (Nanjing, China), and the experimental steps were executed in accordance with the manufacturer's directions. The testosterone test kit was equilibrated at room temperature for half hour an before use. The blank wells were only added with chromogen A, B, and stop solution for zero adjustment. Then, 50 μl of diluted standard and 50 μl of biotinylated antigen working solution were added to each standard well. Next, 50 μl of sample was added to the sample wells, followed by 50 μl of biotinylated antigen working solution. The plate was gently shaken, covered with a sealing film, and incubated in a 37℃ incubator for 30 min. The sealing film was carefully removed, the liquid was discarded, and the wells were flicked dry. Each well was filled with wash solution, and standing after for 30 s, the liquid was discarded. This process was repeated five times, and the plate was patted dry. After the first wash, 50 μl of streptavidin-HRP was added to the zero wells, standard wells, and sample wells. The plate was gently shaken, covered with a sealing film, and incubated in a 37℃ incubator for 30 min. After the second wash, 50 μl of chromogen A was added to each well, followed by 50 μl of chromogen B. The plate was gently shaken to mix, and the color development was carried out at 37℃ in the dark for 10 min. Then, 50 μl of stop solution was added to each well to terminate the reaction. The absorbance (OD value) of each well was measured at 450 nm wavelength in sequence, with the blank well used for zero adjustment. The measurement was completed within 10 min after the addition of the stop solution. Finally, a standard curve was established, and the testosterone concentration was calculated.

Oxidative stress assessment

Fresh testicular tissue was collected and cleansed with pre-cooled PBS to remove impurities. Then, 0.1 g of the tissue was weighed, minced, and homogenized mechanically on ice using 9 times the volume of normal saline (weight g: volume ml = 1:9). The homogenate was subsequently centrifuged at 12,000 rpm for 3 min at 4°C, and the supernatant was harvested for subsequent analysis. Then, according to the method provided by Nanjing Jiancheng Bioengineering Institute (Nanjing, China), oxidative and antioxidant indicators were detected, including catalase (CAT), total antioxidant capacity (T-AOC), glutathione (GSH), malondialdehyde (MDA), and hydrogen peroxide (H2O2). The detection wavelengths for CAT, T-AOC, GSH, MDA, and H2O2 were 405 nm, 520 nm, 412 nm, 532 nm, and 405 nm, respectively. The optical density (OD) values were measured at the selected wavelengths using a microplate reader. Finally, the results were calculated according to the methods provided in the kit instructions. Additionally, detailed information about the kit is shown in Supplementary Table 1. Additionally, this study assessed changes in ROS levels in the testes using the kit provided by Nanjing Jiancheng Bioengineering Institute (Nanjing, China). Please refer to the instructions for specific methods.

TUNEL staining detection

The TUNEL assay was utilized to detect variations in apoptotic levels in the testes, and the procedure strictly adhered to the instructions provided with the kit from Sevier Biotechnology Company (Wuhan, China). After the paraffin sections were soaked in xylene and ethanol respectively, we used proteinase K working solution to treat the sections for 20 min, Following this, phosphate-buffered saline (PBS) was used to wash the sections twice, ensuring the removal of any leftover enzymes. Once drying was complete, the sections were soaked in 50 μl of TUNEL reaction mixture for 60 min. Additionally, select some sections, add 100μ L of diluted DNase I working solution, and incubate for 15 min. Subsequently, DAPI fluorochrome was used to stain the cell nuclei, and all sections were incubated in a dark chamber for 8 min to ensure adequate staining. the sections were mounted with a mounting solution containing an anti-fluorescence quencher. After that, photos were taken using a fluorescence microscope (Nikon, Japan) at 100×magnification, and the apoptotic index was analyzed to assess the changes in the level of apoptosis in the testes.

Determination of the mRNA expression

Total RNA from the testis samples was isolated using Trizol reagent (Invitrogen, USA). The RNA's concentration and purity levels were determined with a UV spectrophotometer (Thermo Fisher Scientific, China). The reaction conditions for reverse transcription were as follows: 20 min at 37°C in a constant temperature water bath, followed by 5 min at 95°C. The specific reverse transcription process was conducted following the protocol provided by Hangzhou Bioer Technology Co., Ltd. (Hangzhou, China). Subsequently, the conventional SYBR Green fluorescent dye method was employed for detection (Roche, USA). We calculated the mRNA relative expression of target genes through an internal reference beta-actin gene. Primers are detailed in Supplementary Table 2.

Immunofluorescence staining

Appropriately sized testicular samples were prepared for paraffin embedding and sectioned into 4 μm thick slices. The sections were placed in a chamber containing an EDTA antigen retrieval solution to facilitate antigen retrieval. Subsequently, blocking was performed by adding bovine serum albumin (BSA) and incubating for 30 min. Afterward, the blocking solution was carefully removed, and the primary antibody was applied to the sections for incubation throughout the night at a temperature of 4°C. The primary antibodies used were as follows: anti-Nrf2 (Bioss; dilution 1:100), anti-HO-1 (Abclonal; dilution 1:100), and anti-IL-6 (Wanlei; dilution 1:200). The next day, a incubation with the secondary antibody (Proteintech; dilution 1:350) was applied to the sections for 50 min. Nuclei were counterstained using DAPI in the dark. An autofluorescence quencher was then applied for 5 min, followed by a thorough rinse with running water for 10 min. Excess moisture was gently blotted away. Ultimately, all sections were examined using a fluorescence microscope (Nikon, Japan) at 200×magnification to assess the intensity of immunofluorescence in the testes, and images of the samples were captured.

Western blot detection

Western blot (WB) technology was utilized to investigate the expression level of specific proteins in testicular tissue. Initially, approximately 30 mg of the tissue sample was finely minced with scissors, followed by homogenization using a homogenizer. Subsequently, 400 μl of lysis buffer (Beyotime, China) was added to the homogenate. The mixture was then centrifuged at 4°C for 10 min, after which the supernatant was carefully collected and reserved for further analysis. Concurrently, the protein concentration of the supernatant was ascertained using the bicinchoninic acid (BCA) (Beyotime, China) assay, a standard quantitative method in protein analysis. subsequently, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was conducted. Following electrophoresis, the proteins were transferred onto a polyvinylidene fluoride (PVDF) membrane using a wet transfer method. Upon completion of the transfer, the PVDF membrane was removed, briefly washed in Tris-buffered saline with Tween-20 (TBST) for 5 min, and then immersed in a blocking solution containing 5 % skim milk powder for 1 h at room temperature. After blocking, the membrane was transferred to an incubation chamber containing the appropriately diluted primary antibody and incubated overnight at 4°C to allow for specific antigen-antibody interactions. The primary antibodies used were as follows: anti-Caspase-3 (Wanlei; dilution 1:300), and anti-IL-6 (Wanlei; dilution 1:500). The next day, the membrane, now incubated with the primary antibody, was subjected to three successive washes with TBST. It was then incubated with a diluted secondary antibody (Abclonal; dilution 1:4000) at room temperature for 2 h. The membrane was prepared for detection using an enhanced chemiluminescence (ECL) (Amersham Biosciences, USA) substrate, and exposure equipment was used to visualize protein bands. The resultant bands were analyzed using ImageJ software.

Differential gene biology analysis

The Metascape website (http://metascape.org) was employed to perform an analysis of differential gene enrichment, serving as a tool for gene set enrichment analysis and functional annotation.Additionally, correlation analysis was carried out using Originpro2021 software. To determine the correlation in statistical analysis for related types, the Pearson method was utilized. The Nrf2/HO-1 signaling pathway markers, oxidative stress indicators, and markers for apoptosis and inflammation were examined through the application of these tools.

Statistical analysis

Experimental data were represented using GraphPad Prism and Origin software for visualization and significance analysis. A quantitative assessment of immunofluorescence staining was performed utilizing ImageJ software. Significant differences among the groups were evaluated using GraphPad Prism version 8.3.0 through one-way ANOVA followed by Tukey's post-hoc test. The experiments were conducted three times, and the results were presented as mean ± standard deviations (SD). The levels of statistical significance are indicated by asterisk and hash as follows: *P < 0.05, **P < 0.01, ***P < 0.001,#P < 0.05, ##P < 0.01, ###P < 0.001.

Results

Ultrastructural analysis of testicular tissue

To investigate the impact of dietary Cd and Se supplementation on testicular integrity, this study employed TEM to assess alterations in testicular morphology. The findings are depicted in Fig. 1, testis samples from the Con, LS, and HS groups showed good structural characteristics, with intact nuclear membranes (black single arrows). Conversely, the group exposed solely to Cd displayed evidence of nuclear membrane fragmentation (red triangle). and perinuclear space widening (blue single arrow). Not surprisingly, the above-mentioned nuclear damage phenomenon also occurred in groups Cd+LS and Cd+HS, however, the nuclear membrane damage in the Cd+HS group was alleviated. Hence, this study showed that Se could partially alleviate Cd-induced damage to testicular ultrastructure in chickens.

Fig. 1.

Fig 1

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Effects of Cd and Se treatment on the ultrastructure of chicken testis. Transmission electron microscopy (TEM) images of testicular tissue across different groups are presented. The nuclear membrane is indicated by 'NM', the nucleus by 'N', red triangle denotes a rupture in the nuclear membrane, and blue arrow signifies the dilation of the perinuclear space.

Effects of Cd and Se on serum testosterone levels in chickens

Testosterone, a critical sex hormone produced in the testicles, plays a significant role in male reproductive function. This study sought to examine the result of dietary Cd and Se supplementation on serum T secretion levels. Radioimmunoassay was utilized to measure the serum T content across all groups. The result was shown in Fig. 2 that supplementation with either the LS group or the HS group had no discernible effect on serum T levels. However, in contrast with the control group, the Cd, Cd+LS, and Cd+HS groups exhibited a marked decrease in serum T (P < 0.001, P < 0.001, P < 0.05). Furthermore, the serum T levels in the Cd+HS group demonstrated a significant increase compared to the Cd group (P < 0.001). These findings indicated that Cd triggered a decrease in serum testosterone secretion levels of chickens, and Se could alleviate the above-mentioned downward trend in serum testosterone.

Fig. 2.

Fig 2

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Effects of Cd and Se treatment on serum testosterone content in chickens. Differences from the control group are denoted by an asterisk (*). Specifically, P values less than 0.05 are marked with *, less than 0.01 with **, and less than 0.001 with ***. Distinctions between the Cd group and other groups are marked with a hash (#). Here, statistical significance is indicated by # for P < 0.05, ## for P < 0.01, and ### for P < 0.001. Results are depicted with mean ± SD.

Effects of Cd and Se on the Nrf2/HO-1 signaling pathway in the testicle

To investigate the potential impact of dietary Cd and Se on the Nrf2/HO-1 signaling pathway in the testis, we used qRT-PCR and immunofluorescence staining to measure the pathway's expression levels, and the results are shown in Fig. 3A–F. Compared to the Con group, the LS, HS, Cd+LS, and Cd+HS groups showed a rise in Nrf2 and HO-1 mRNA expression levels. In contrast, the Cd group displayed a significant reduction in Nrf2 and HO-1 mRNA expression levels. Furthermore, in contrast to the Cd group, the Cd+LS and Cd+HS groups showed an upregulation of Nrf2 and HO-1 mRNA expression levels. Following Se treatment, the expression of Nrf2 and HO-1 was primarily increased in the testicular interstitium. In addition, Cd exposure led to a pronounced reduction in the immunofluorescence intensity of testicular Nrf2 and HO-1 compared to the Con group (P < 0.001, P < 0.05). It is noteworthy that the Cd+HS group displayed a marked elevation in Nrf2 and HO-1 fluorescence intensity when contrasted with the Cd group (P < 0.001), indicating a protective role of high-Se supplementation against Cd-induced suppression of the Nrf2/HO-1 signaling pathway in testicles.

Fig. 3.

Fig 3

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The impact of Cd and Se treatment on Nrf2/HO-1 signaling pathway in chicken testicles. (A-B) The expression levels of Nrf2 and HO-1 were evaluated using immunofluorescence staining (Scale bar = 100 μm). (C-D) The relative fluorescence intensities of Nrf2 and HO-1 were quantified. (E-F) Relative mRNA expression levels of Nrf2 and HO-1. Differences from the control group are denoted by an asterisk (*). Specifically, P values less than 0.05 are marked with *, less than 0.01 with **, and less than 0.001 with ***. Distinctions between the Cd group and other groups are marked with a hash (#). Here, statistical significance is indicated by # for P < 0.05, ## for P < 0.01, and ### for P < 0.001. Results are depicted with mean ± SD.

The role of Cd and Se in modulating oxidative stress indicators and ROS levels in the testicle

Oxidative stress is a significant inducer of testicular damage. This investigation sought to examine the influences of dietary Cd and Se on testicular oxidative stress by assessing ROS levels and related indicators. As depicted in Fig. 4A H, there was an absence of significant differences in the levels of ROS, T-AOC, GSH, H2O2, MDA, and CAT between the LS and Con groups. However, in the Cd group, significant downregulation occurred in the levels of T-AOC, GSH, and CAT (P < 0.001), along with a significant increase in ROS levels and the levels of H2O2 and MDA (P < 0.001). With high selenium supplementation, the Cd+HS group demonstrated a significant boost in T-AOC, GSH, and CAT levels when contrasted with the Cd group (P < 0.001). Additionally, the ROS levels, as well as the levels of H2O2 and MDA, were markedly lower in the Cd+LS and Cd+HS groups than in the Cd group. As shown in Fig. 4I–K, compared with the Con group, the mRNA expression levels of CAT, SOD1 and SOD2 were significantly reduced in the Cd, Cd+LS and Cd+HS groups (P < 0.001, P < 0.001, P < 0.001). However, the Cd+LS and Cd+HS groups could alleviate the above-mentioned decrease in mRNA expression. Thus, our findings suggested that Se exerted an inhibitory effect on Cd-induced oxidative stress in testicles.

Fig. 4.

Fig 4

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Effects of Cd and Se treatment on oxidative stress in chicken testicles. (A-E) Assessment of oxidative stress-related markers (CAT, H2O2, GSH, MDA, T-AOC). (F) Measurement of ROS levels. (G-H) Heat map of different enzymes and ROS content, and the content gradually increases from orange to green. (I-K) The mRNA expression levels of CAT, SOD1 and SOD2. Differences from the control group are denoted by an asterisk (*). Specifically, P values less than 0.05 are marked with *, less than 0.01 with **, and less than 0.001 with ***. Distinctions between the Cd group and other groups are marked with a hash (#). Here, statistical significance is indicated by # for P < 0.05, ## for P < 0.01, and ### for P < 0.001. Results are depicted with mean ± SD.

Effects of Cd and Se on apoptotic signaling pathway in the testicle

To assess the impact of dietary Cd and Se supplementation on apoptotic signaling pathway, we employed qRT-PCR, Western blot, and TUNEL staining to evaluate testicular apoptosis levels. As shown in Fig. 5A and B, TUNEL assay results demonstrated a significant increase in the apoptotic cell index in the Cd, Cd+LS, and Cd+HS groups in comparison with the Con group (P < 0.001, P < 0.001, P < 0.05). It is worth noting that apoptotic cells were primarily localized within the lumen of the seminiferous tubules. Nonetheless, groups treated with Cd+LS and Cd+HS showed a marked decline in the trend of elevated apoptotic cell counts compared to the Cd group (P < 0.001, P < 0.001). As presented in Fig. 5E–I, our results showed no noteworthy variations in the mRNA expression levels of Caspase-3, Bcl-2, and Cyt-c among the Con, LS, and HS groups. However, in the Cd group, a significant upregulation was detected in the mRNA levels of Caspase-3, Bax, Cyt-c, and Caspase-9 (P < 0.001), while a significant reduction was observed in Bcl-2 mRNA expression levels (P < 0.001). The Cd+LS and Cd+HS groups showed a significant lowering of mRNA expression levels for Caspase-3, Bax, Caspase-9, and Cyt-c in comparison with the Cd group (P < 0.001, P < 0.001). However, in contrast with the Cd group, the mRNA expression levels of Bcl-2 elevated significantly in both the Cd+LS and Cd+HS groups (P < 0.001, P < 0.001). Furthermore, as illustrated in Fig. 5C and D, there was a notable increase in the protein expression levels of Caspase-3 in the HS, Cd, Cd+LS, and Cd+HS groups when compared to the Con group. Notably, the protein expression of Caspase-3 in the Cd+HS group exhibited a significant reduction relative to the Cd group (P < 0.001). These results indicated that Cd triggered the apoptotic signaling pathway in the testis and increased the number of apoptotic cells, whereas Se had an antagonistic effect on testicular cell apoptosis.

Fig. 5.

Fig 5

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Effects of Cd and Se treatment on chicken testicular cell apoptosis (A) TUNEL staining assessment (Scale bar = 200 μm). (B) Quantitative analysis of the apoptosis index. (C-D) Protein bands and relative protein expression levels of Caspase3. (E-I) Relative mRNA expression levels of apoptosis signaling pathway markers (Caspase3, Bcl-2, Cyt-c, Caspase-9, Bax). Differences from the control group are denoted by an asterisk (*). Specifically, P values less than 0.05 are marked with *, less than 0.01 with **, and less than 0.001 with ***. Distinctions between the Cd group and other groups are marked with a hash (#). Here, statistical significance is indicated by # for P < 0.05, ## for P < 0.01, and ### for P < 0.001. Results are depicted with mean ± SD.

The impact of Cd and Se on inflammation-related indicators in the testicle

To establish the occurrence of an inflammatory response in the testicle, we examined the expression levels of inflammation-related indicators. As depicted in Fig. 6A and B, immunofluorescence findings revealed no significant changes in the fluorescence intensity of IL-6 in the LS and HS groups compared to the Con group. In contrast, a notable increase in fluorescence intensity for IL-6 was observed in both the Cd and Cd+LS groups (P < 0.001, P < 0.001). The fluorescence intensity of IL-6 was significantly diminished in both the Cd+LS and Cd+HS groups compared with the Cd group (P < 0.001, P < 0.001). As presented in Fig. 6C and D, a significant elevation in IL-6 protein expression was observed in the Cd group relative to the Con group (P < 0.001). The application of either LS or HS in conjunction with Cd treatment resulted in a significant reduction in IL-6 protein levels compared to the Cd group alone. Additionally, as depicted in Fig. 6E–H, the results showed that no significant alterations were observed in the mRNA expression levels of IL-6, TNF-α, and IL-2 between the Con group and the LS group. However, the mRNA expression levels of IL-6, TNF-α, IL-2, and IL-1β in the Cd, LS+Cd, and HS+Cd groups were considerably higher when contrasted with the Con group (P < 0.001, P < 0.001, P < 0.001). Compared to the Cd group, the mRNA expression levels of IL-6, TNF-α, IL-2, and IL-1β were significantly reduced in both the Cd+LS and Cd+HS groups (P < 0.001, P < 0.001). The research suggested that Cd triggered testicular inflammation, while Se provided a protective effect against these inflammatory response.

Fig. 6.

Fig 6

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Impact of Se and Cd treatment on testicular inflammatory signaling pathway in chickens (A) Immunofluorescence staining for IL-6 (Scale bar = 100 μm). (B) Quantitative analysis of IL-6 relative fluorescence intensity. (C-D) Relative protein expression levels of IL-6 and protein bands. (E-H) Relative mRNA expression levels of inflammatory markers (IL-6, TNF-α, IL-2, IL-1β). Differences from the control group are denoted by an asterisk (*). Specifically, P values less than 0.05 are marked with *, less than 0.01 with **, and less than 0.001 with ***. Distinctions between the Cd group and other groups are marked with a hash (#). Here, statistical significance is indicated by # for P < 0.05, ## for P < 0.01, and ### for P < 0.001. Results are depicted with mean ± SD.

Differential gene analysis results

As depicted in Fig. 7A, 11 pathways were discovered in enriched pathway terms. WP4630: Measles virus infection, GO:0042542: response to hydrogen peroxide, R-HSA-449147: Signaling by Interleukins, WP5434: Pathways in cancer, WP3287: Overview of nanoparticle effects, WP2328: Allograft rejection, WP4008: NO cGMP PKG mediated neuroprotection, WP1533: Vitamin B12 metabolism, WP4891: COVID 19 adverse outcome pathway, R-HSA-2151201: Transcriptional activation of mitochondrial biogenesis, GO:1903706: regulation of hemopoiesis. In Fig. 7B, nodes of various colors were identified, with each color representing a distinct term. Moreover, the nodes' sizes corresponded to the quantity of genes. In an effort to better understand the interplay among Nrf2/HO-1 markers, the oxidative stress response, and indicators of apoptosis and inflammation, we carried out a correlation analysis. Fig. 7C indicated that the Nrf2/HO-1/oxidative stress signaling pathway was negatively correlated with pathways that were involved in apoptosis and inflammation.

Fig. 7.

Fig 7

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Comprehensive analysis of differential genes in chicken testis. (A) Bar graph of enriched terms for different expressed genes, terms were colored by p-values. (B) Network of enriched terms colored by cluster ID and p-values. (C) Correlation analysis among the Nrf2/HO-1 signaling pathway markers, oxidative stress indicators, and markers for apoptosis and inflammation in chicken testis, and the use of different colors denotes various correlations.

Discussion

Cd is a well-known reproductive disruptor present in the environment, with a particular impact on the testicles. Even at low levels, Cd can induce toxic effects in the testicles. Research demonstrated that Cd could lead to testicular degeneration and increase sperm abnormality rate in rats (Venditti et al., 2021; Li et al., 2024b). A study showed that Cd decreased testosterone levels in the blood and caused structural abnormalities of the blood testicular barrier (BTB) in mice, affecting testicular development and sperm production (Wang et al., 2023a; Fang et al., 2020). Se was identified as a substance that could counteract the harmful effects of Cd, thereby safeguarding the testicles from damage (Zhang et al., 2024b). Our research explored the adverse effects of Cd on chicken testicles and assessed whether Se could mitigate the detrimental effects of Cd. The results indicated that Cd caused nuclear membrane damage and oxidative stress in chicken testicles. Furthermore, Cd suppressed the Nrf2/HO-1 signaling pathway and activated the apoptosis signaling pathway and inflammatory response in chicken testicles. Conversely, Se mitigated the damage to the chicken testicle and prevented the downregulation of the Nrf2/HO-1 signaling pathway caused by Cd. This study also revealed that Se effectively counteracted the excessive oxidative stress and mitigated the expression of factors related to apoptosis and inflammation in chicken testicles after Cd stimulation.

Under normal conditions, the oxidative enzyme system and the antioxidant enzyme system maintain a delicate balance in the testicle (Xiao et al., 2024), which is essential for normal testicular development and prevention of oxidative damage. However, Cd can disrupt this equilibrium, leading to oxidative stress in the testicle (Pu et al., 2023). Specifically, it was shown that Cd could elevate the levels of the oxidative marker MDA and diminish the levels of CAT and GSH in rat testes, indicating the occurrence of oxidative stress (Djuric et al., 2015). In line with these findings, our study confirmed that the levels of ROS, H2O2, and MDA were elevated in chicken testicles after Cd stimulation, while concurrently reducing the levels of CAT, GSH, and T-AOC. These results underscored the pro-oxidative impact of Cd on avian testicular health. Emerging scientific evidence had established an antagonistic relationship between Se and Cd, with Se being recognized for its potent antioxidant properties (Wang et al., 2023b). Se could effectively neutralize free radicals and peroxides generated by Cd, thus mitigating oxidative stress. For instance, selenium yeast had been shown to counteract Cd-induced oxidative damage in the chicken liver by upregulating the expression of SOD, GSH-Px, and CAT (Wang et al., 2020). Consistent with previous studies, our study demonstrated that co-treatment with Se and Cd markedly improved the levels of T-AOC, GSH, and CAT in chicken testicles, while concurrently prompting a marked reduction in the levels of ROS, H2O2, and MDA. These events showed that Se demonstrated a protective role against oxidative stress induced by Cd in chicken testicles.

The Nrf2/HO-1 signaling pathway is recognized as a pivotal endogenous defense mechanism in animals, particularly noted for its role in combatting oxidative stress (Hassanein et al., 2023; Bai et al., 2024). Without external stimulation, Nrf2 and HO-1 are typically expressed at low levels. However, research has indicated that Cd not only suppresses the expression of these two proteins but also exacerbates oxidative stress (Alruhaimi et al., 2023). Studies on hepatic injury in rats demonstrated that Cd accumulation could lead to a suppression of Nrf2 and HO-1 expression, consequently, this increased the levels of oxidative stress (Fang et al., 2021). Furthermore, research confirmed that Cd exerted a suppressive influence on the Nrf2/HO-1 signaling pathway, resulting in heightened oxidative stress and subsequent renal damage (Das et al., 2019). Consistent with these findings, our investigation into chicken testicles revealed that Cd led to a downregulation of Nrf2 and HO-1, coinciding with an escalation in oxidative stress. The supplementation of Se in the diet was demonstrated to stimulate the Nrf2/HO-1 signaling pathway, which could improve the antioxidant system's effectiveness and promote cellular survival. In the testicular tissue, the main cells present in the testicular interstitium were the leydig cells. These leydig cells were the key cell types responsible for the synthesis and secretion of androgens, providing essential endocrine support for spermatogenesis. In our study, following Se treatment, the expression of Nrf2/HO-1 was primarily increased in leydig cells. This may have been because leydig cells were able to absorb Se elements more effectively, thereby possessing a stronger capacity for antioxidant defense. As an antioxidant, Se likely activated the Nrf2 signaling pathway, thereby enhancing the antioxidant capacity of leydig cells and protecting them from oxidative stress damage. This increase in expression helped maintain the normal function of leydig cells and was beneficial for protecting the process of spermatogenesis from oxidative stress interference. Studies on renal toxicity in rats suggested that varying doses of Se could lower oxidative stress by stimulating the Nrf2/HO-1/NQO1 cascade (Hu et al., 2023). Additionally, the activation of the Nrf2/HO-1 signaling pathway by Se nanoparticles had been linked to a reduction in cellular oxidative stress resulting from H2O2 exposure (Zhu et al., 2023). In comparison with the group subjected to Cd alone, we observed in our study that co-treatment with Se and Cd increased the production of Nrf2 and HO-1, coupled with a decrease in oxidative stress levels. The findings indicated that Se diminished oxidative stress caused by Cd in chicken testicles through the induction of the Nrf2/HO-1 signaling pathway.

Growing evidence suggests that cell damage such as apoptosis, necroptosis, and inflammation in different tissues or organs is closely related to oxidative stress (Liu et al., 2022; Li et al., 2024). In reality, numerous pollutants, including the heavy metal Cd, can inflict cellular harm via the induction of oxidative stress (Ju et al., 2024). Related research reported that Cd triggered oxidative stress in duck renal tubular epithelial cells, subsequently initiating apoptosis (Zhuang et al., 2019). Moreover, oxidative stress was implicated in regulating inflammatory responses triggered by Cd exposure in the small intestine of pigs (Chen et al., 2022). Aligning with prior findings, our study identified that oxidative stress contributed to apoptosis and inflammation induced by Cd in chicken testicles. In this study, it was found that apoptotic cells were primarily localized within the seminiferous tubules. The reason for this phenomenon may have been related to the higher sensitivity of germ cells and sertoli cells to oxidative stress. These cells underwent complex cell division processes during spermatogenesis and had higher requirements for their survival environment, such as the levels of oxidative stress or the demand for nutrients. These cells were also more prone to damage during development, which could lead to cell apoptosis. Therefore, this finding provided a certain reference value for future in-depth research into the oxidative stress responses and apoptosis mechanisms of different testicular cells. An extensive body of research had established that Se was a potent scavenger of deleterious free radicals and possesses significant antioxidant properties. Se alleviated heavy metal lead-induced apoptosis of sheep leydig cells by reducing oxidative stress (Shi et al., 2022). Moreover, a deficiency in Se had been linked to heightened oxidative stress in the myocardium of calf, resulting in inflammation and apoptosis (Lei et al., 2023). our study showed that the co-treatment with Se and Cd led to lowered levels of apoptosis and inflammation compared to the group treated with Cd alone. These results revealed that Se could counteract Cd-mediated apoptosis and inflammation through oxidative stress. In conjunction with previous research, we demonstrated that the Nrf2/HO-1 signaling pathway, activated by Se, could reduce the oxidative stress induced by Cd, thereby alleviating apoptosis and inflammation in chicken testicles.

Conclusion

In summary, the present study delineated a novel mechanistic insight into how Se counteracted Cd-induced testicular damage in chickens. Se potentially ameliorated Cd-induced apoptosis and inflammation in chicken testicles by inhibiting oxidative stress via promoting the Nrf2/HO-1 signaling pathway. Overall, our findings provide new evidence for the application of Se as an effective antidote for Cd-induced male reproductive damage.

Disclosures

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Acknowledgments

The study was supported by the Basic Scientific Research Project of Provincial Universities in Heilongjiang Province (145409529).

Footnotes

Supplementary material associated with this article can be found, in the online version, at doi:10.1016/j.psj.2025.104990.

Appendix. Supplementary materials

mmc1.docx (15.9KB, docx)

References

  1. Alruhaimi R.S., Hassanein E.H.M., Abd El-Aziz M.K., Siddiq Abduh M., Bin-Ammar A., Kamel E.M., Mahmoud A.M., Mahmoud A.M. The melatonin receptor agonist agomelatine protects against acute pancreatitis induced by cadmium by attenuating inflammation and oxidative stress and modulating Nrf2/HO-1 pathway. Int. Immunopharmacol. 2023;124 doi: 10.1016/j.intimp.2023.110833. [DOI] [PubMed] [Google Scholar]
  2. Bai Y., Sui R., Zhang L., Bai B., Zhu Y., Jiang H. Resveratrol improves cognitive function in post-stroke depression rats by repressing inflammatory reactions and oxidative stress via the Nrf2/HO-1 pathway. Neuroscience. 2024;541:50–63. doi: 10.1016/j.neuroscience.2024.01.017. [DOI] [PubMed] [Google Scholar]
  3. Bautista C.J., Arango N., Plata C., Mitre-Aguilar I.B., Trujillo J., Ramírez V. Mechanism of cadmium-induced nephrotoxicity. Toxicology. 2024;502 doi: 10.1016/j.tox.2024.153726. [DOI] [PubMed] [Google Scholar]
  4. Binmahfouz L.S., Hassanein E.H.M., Bagher A.M., Hareeria R.H., Alamri Z.Z., Algandaby M.M., Abdel-Daim M.M., Abdel-Naim A.B. Berberine alleviates chlorpyrifos-induced nephrotoxicity in rats via modulation of Nrf2/HO-1 axis. Heliyon. 2024;10:e25233. doi: 10.1016/j.heliyon.2024.e25233. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen X., Bi M., Yang J., Cai J., Zhang H., Zhu Y., Zheng Y., Liu Q., Shi G., Zhang Z. Cadmium exposure triggers oxidative stress, necroptosis, Th1/Th2 imbalance and promotes inflammation through the TNF-α/NF-κb pathway in swine small intestine. J. Hazard. Mater. 2022;421 doi: 10.1016/j.jhazmat.2021.126704. [DOI] [PubMed] [Google Scholar]
  6. Das S., Dewanjee S., Dua T.K., Joardar S., Chakraborty P., Bhowmick S., Saha A., Bhattacharjee S., De Feo V. Carnosic acid attenuates cadmium induced nephrotoxicity by inhibiting oxidative stress, promoting Nrf2/HO-1 signalling and impairing TGF-β1/Smad/Collagen IV signalling. Molecules. 2019;24:4176. doi: 10.3390/molecules24224176. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Djuric A., Begica A., Gobeljica B., Stanojevic I., Ninkovic M., Vojvodic D., Pantelic A., Zebic G., Prokic V., Dejanovic B., Stojanovic I., Pavlica M., Djukic D., Saso L., Djurdjevic D., Pavlovic M., Topic A., Vujanovic D., Stevnovic I., Djukic M. Oxidative stress, bioelements and androgen status in testes of rats subacutely exposed to cadmium. Food Chem. Toxicol. 2015;86:25–33. doi: 10.1016/j.fct.2015.09.004. [DOI] [PubMed] [Google Scholar]
  8. Fang J., Yin H., Yang Z., Tan M., Wang F., Chen K., Zuo Z., Shu G., Cui H., Ouyang P., Guo H., Chen Z., Huang C., Geng Y., Liu W. Vitamin E protects against cadmium-induced sub-chronic liver injury associated with the inhibition of oxidative stress and activation of Nrf2 pathway. Ecotoxicol. Environ. Saf. 2021;208 doi: 10.1016/j.ecoenv.2020.111610. [DOI] [PubMed] [Google Scholar]
  9. Fang Y., Xiang Y., Lu X., Dong X., Zhang J., Zhong S., Zhong S. Epigenetic dysregulation of Mdr1b in the blood-testis barrier contributes to dyszoospermia in mice exposed to cadmium. Ecotoxicol. Environ. Saf. 2020;190 doi: 10.1016/j.ecoenv.2019.110142. [DOI] [PubMed] [Google Scholar]
  10. Fan R., Hu P., Wang Y., Lin H., Su K., Feng X., Wei L., Yang F., Yang F. Betulinic acid protects mice from cadmium chloride-induced toxicity by inhibiting cadmium-induced apoptosis in kidney and liver. Toxicol. Lett. 2018;299:56–66. doi: 10.1016/j.toxlet.2018.09.003. [DOI] [PubMed] [Google Scholar]
  11. Fouad A.A., Abdel-Aziz A.M., Hamouda A.A.H. Diacerein downregulates NLRP3/caspase-1/IL-1β and IL-6/STAT3 pathways of inflammation and apoptosis in a rat model of cadmium testicular toxicity. Biol. Trace. Elem. Res. 2019;195:499–505. doi: 10.1007/s12011-019-01865-6. [DOI] [PubMed] [Google Scholar]
  12. Ge J., Guo K., Zhang C., Talukder M., Lv M., Li J., Li J. Comparison of nanoparticle-selenium, selenium-enriched yeast, and sodium selenite on the alleviation of cadmium-induced inflammation via NF-kB/iκb pathway in heart. Sci. Total Environ. 2021;773 doi: 10.1016/j.scitotenv.2021.145442. [DOI] [PubMed] [Google Scholar]
  13. Hassanein E.H.M., Ibrahim I.M., Abd-alhameed E.K., Sharawi Z.W., Jaberd F.A., Althagaf H.S., Al-Salem A.A. Nrf2/HO-1 as a therapeutic target in renal fibrosis. Life Sci. 2023;334 doi: 10.1016/j.lfs.2023.122209. [DOI] [PubMed] [Google Scholar]
  14. Hu Y., Yan Z., He Y., Li Y., Li M., Li Y., Zhang D., Zhao Y., Ommati M.M., Wang J., Huo M., Wang J. Ameliorative effects of different doses of selenium against fluoride-triggered apoptosis and oxidative stress-mediated renal injury in rats through the activation of Nrf2/HO-1/NQO1 signaling pathway. Food Chem. Toxicol. 2023;174 doi: 10.1016/j.fct.2023.113647. [DOI] [PubMed] [Google Scholar]
  15. Hughes D.J., Schomburg L., Jenab M., Biessy C., M´eplan C., Moskal A., Mukhtar M. Prediagnostic selenium status, selenoprotein gene variants and association with breast cancer risk in a European cohort study. Free Radic. Bio. Med. 2023;209:381–393. doi: 10.1016/j.freeradbiomed.2023.10.401. [DOI] [PubMed] [Google Scholar]
  16. Hou T., Zhu L., Wang Y., Peng L. Oxidative stress is the pivot for PM2.5-induced lung injury. Food Chem. Toxicol. 2024;184 doi: 10.1016/j.fct.2023.114362. [DOI] [PubMed] [Google Scholar]
  17. Ibrahim R.E., Elshobaky G., ElHady M., Abdelwarith A.A., Younis E.M., Rhoumad N.R.…Abdel Rahman A.N. Nelumbo nucifera synthesized selenium nanoparticles modulate the immune-antioxidants, biochemical indices, and pro/anti-inflammatory cytokines pathways in Oreochromis niloticus infected with Aeromonas veronii. Fish Shellf. Immun. 2024;144 doi: 10.1016/j.fsi.2023.109287. [DOI] [PubMed] [Google Scholar]
  18. Ju Z., Ndandala C.B., Lei Y., Shija V.M., Luo J., Wang P., Wen C., Liang H. Cadmium-induced oxidative stress, histopathology, and transcriptomic changes in the hepatopancreas of Fenneropenaeus merguiensis. Aquacult. Rep. 2024;36 [Google Scholar]
  19. Lei L., Mu J., Zheng Y., Liu Y. Selenium deficiency-induced oxidative stress causes myocardial injury in calves by activating inflammation, apoptosis, and necroptosis. Antioxidants. 2023;12:229. doi: 10.3390/antiox12020229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Liu X., Zhang Y., Sun X., Zhang W., Shi X., Xu S. Di-(2-ethyl hexyl) phthalate induced oxidative stress promotes microplastics mediated apoptosis and necroptosis in mice skeletal muscle by inhibiting PI3K/AKT/mTOR pathway. Toxicology. 2022;474 doi: 10.1016/j.tox.2022.153226. [DOI] [PubMed] [Google Scholar]
  21. Li X., Liu J., Wu S., Zheng W., Li H., Bao S., Chen Y., Guo X., Zhang L., Ge R. In utero single low-dose exposure of cadmium induces rat fetal Leydig cell dysfunction. Chemosphere. 2018;194:57–66. doi: 10.1016/j.chemosphere.2017.11.159. [DOI] [PubMed] [Google Scholar]
  22. Li Y., Zhang Y., Feng R., Zheng P., Huang H., Zhou S., Ji W., Huang F., Liu H., Zhang G. Cadmium induces testosterone synthesis disorder by testicular cell damage via TLR4/MAPK/NF-κb signaling pathway leading to reduced sexual behavior in piglets. Ecotoxicol. Environ. Saf. 2022;233 doi: 10.1016/j.ecoenv.2022.113345. [DOI] [PubMed] [Google Scholar]
  23. Li Y., Wang S., Wang Y. Cadmium exposure induces oxidative stress-mediated necroptosis via TLR4/NF-κb signaling pathway in pig epididymis. Environ. Pollut. 2024;366 doi: 10.1016/j.envpol.2024.125514. [DOI] [PubMed] [Google Scholar]
  24. Li Y., Wang H., Wang Y. Cadmium exposure induces inflammation through oxidative stress-mediated activation of the NF‑κb signaling pathway and causes heat shock response in a piglet testis. Biol. Trace. Elem. Res. 2024:1–17. doi: 10.1007/s12011-024-04470-4. [DOI] [PubMed] [Google Scholar]
  25. Liang Y., Jiang L., Hu M., Luo X., Cheng T., Wang Y. Tea tree oil inhibits hydrogen sulfide-induced oxidative damage in chicken lungs through CYP450s/ROS pathway. Poult. Sci. 2024 doi: 10.1016/j.psj.2024.103860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pu W., Chu X., Huang G., Cui T., Huang B., Dai X., Zhang C. The activated ATM/AMPK/mTOR axis promotes autophagy in response to oxidative stress-mediated DNA damage co-induced by molybdenum and cadmium in duck testes. Environ. Pollut. 2023;316 doi: 10.1016/j.envpol.2022.120574. [DOI] [PubMed] [Google Scholar]
  27. Shi L., Wang X., Duan Y., Li K., Ren Y. Antagonistic effects of selenium on lead-induced oxidative stress and apoptosis of Leydig cells in sheep. Theriogenology. 2022;185:43–49. doi: 10.1016/j.theriogenology.2022.03.023. [DOI] [PubMed] [Google Scholar]
  28. Suhani I., Sinha S., Srivastava V., Singh R.P. Impact of cadmium pollution on food safety and human health. Curr. Opin. Toxicol. 2021;27:1–7. [Google Scholar]
  29. Venditti M., Ben Rhouma M., Romano M.Z., Messaoudi I., Reiter R.J., Minucci S. Evidence of melatonin ameliorative effects on the blood-testis barrier and sperm quality alterations induced by cadmium in the rat testis. Ecotoxicol. Environ. Saf. 2021;226 doi: 10.1016/j.ecoenv.2021.112878. [DOI] [PubMed] [Google Scholar]
  30. Valipour J., Taghizadeh F., Esfahani R., Ramesh M., Rastegar T. Role of nuclear factor erythroid 2-related factor 2 (Nrf2) in female and male fertility. Heliyon. 2024;10:e29752. doi: 10.1016/j.heliyon.2024.e29752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wang Y., Wu J., Zhang M., Yang H., Li M., Jia D., Wang R., Zhou W., Liu H., Hu Y., Yao Y., Liu Y., Ji Y. Cadmium exposure during puberty damages testicular development and spermatogenesis via ferroptosis caused by intracellular iron overload and oxidative stress in mice. Environ. Pollut. 2023;325 doi: 10.1016/j.envpol.2023.121434. [DOI] [PubMed] [Google Scholar]
  32. Wang M., Wang Y., Wang S., Hou L., Cui Z., Li Q., Huang H. Selenium alleviates cadmium-induced oxidative stress, endoplasmic reticulum stress and programmed necrosis in chicken testes. Sci. Total Environ. 2023;863 doi: 10.1016/j.scitotenv.2022.160601. [DOI] [PubMed] [Google Scholar]
  33. Wang Y., Chen H., Chang W., Chen R., Xu S., Tao D. Protective effects of selenium yeast against cadmium-induced necroptosis via inhibition of oxidative stress and MAPK pathway in chicken liver. Ecotoxicol. Environ. Saf. 2020;206 doi: 10.1016/j.ecoenv.2020.111329. [DOI] [PubMed] [Google Scholar]
  34. Wang Z., Wang Y., Lü J., Li T., Li S., Nie M., Shi G., Zhao X. Silicon and selenium alleviate cadmium toxicity in Artemisia selengensis Turcz by regulating the plant-rhizosphere. Environ. Res. 2024;252 doi: 10.1016/j.envres.2024.119064. [DOI] [PubMed] [Google Scholar]
  35. Wan N., Xu Z., Liu T., Min Y., Li S. Ameliorative effects of selenium on cadmium-induced injury in the chicken ovary: mechanisms of oxidative stress and endoplasmic reticulum stress in cadmium-induced apoptosis. Biol. Trace. Elem. Res. 2017;184:463–473. doi: 10.1007/s12011-017-1193-x. [DOI] [PubMed] [Google Scholar]
  36. Xiao Y., Liu R., Tang W., Yang C., Yang C. Cantharidin-induced toxic injury, oxidative stress, and autophagy attenuated by Astragalus polysaccharides in mouse testis. Reprod. Toxicol. 2024;123 doi: 10.1016/j.reprotox.2023.108520. [DOI] [PubMed] [Google Scholar]
  37. Yang X., Fang Y., Hou J., Wang X., Li J., Li S., Zheng X., Liu Y., Zhang Z. The heart as a target for deltamethrin toxicity: inhibition of Nrf2/HO-1 pathway induces oxidative stress and results in inflammation and apoptosis. Chemosphere. 2022;300 doi: 10.1016/j.chemosphere.2022.134479. [DOI] [PubMed] [Google Scholar]
  38. Zhuang J., Nie G., Yang F., Dai X., Cao H., Xing C., Hu G., Zhang C. Cadmium induces cytotoxicity through oxidative stress-mediated apoptosis pathway in duck renal tubular epithelial cells. Toxicol. In Vitro. 2019;61 doi: 10.1016/j.tiv.2019.104625. [DOI] [PubMed] [Google Scholar]
  39. Zhang Z., Zhang Q., Li M., Xu J., Wang J., Li M., Wei L., Lv Q., Chen X., Wang Y., Liu Y. SeMet attenuates AFB1-induced intestinal injury in rabbits by activating the Nrf2 pathway. Ecotoxicol. Environ. Saf. 2022;239 doi: 10.1016/j.ecoenv.2022.113640. [DOI] [PubMed] [Google Scholar]
  40. Zhang Z., Wang Q., Gao X., Tang X., Xu H., Wang W., Lei X. Reproductive toxicity of cadmium stress in male animals. Toxicology. 2024;504 doi: 10.1016/j.tox.2024.153787. [DOI] [PubMed] [Google Scholar]
  41. Zhang L., Shi W.-Y., Zhang L.-L., Sha Y., Xu J.-Y., Shen L.-C., Li Y.-H., Yuan L.-X., Qin L.-Q. Effects of selenium-cadmium co-enriched cardamine hupingshanensis on bone damage in mice. Ecotoxicol. Environ. Saf. 2024;272 doi: 10.1016/j.ecoenv.2024.116101. [DOI] [PubMed] [Google Scholar]
  42. Zhu M., Zhang Y., Zhang C., Chen L., Kuang Y. Rutin modified selenium nanoparticles reduce cell oxidative damage induced by H2O2 by activating Nrf2/HO-1 signaling pathway. J. Biomater. Appl. 2023;38:109–121. doi: 10.1177/08853282231182765. 37. [DOI] [PubMed] [Google Scholar]

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