Skip to main content
NCBI home page
Toxicology Research logoLink to Toxicology Research
. 2024 May 9;13(3):tfae072. doi: 10.1093/toxres/tfae072

Lead-induced liver fibrosis and inflammation in mice by the AMPK/MAPKs/NF-κB and STAT3/TGF-β1/Smad2/3 pathways: the role of Isochlorogenic acid a

Jun-Tao Guo 1,#, Han-Yu Li 2,#, Chao Cheng 3, Jia-Xue Shi 4, Hai-Nan Ruan 5, Jun Li 6, Chan-Min Liu 7,✉
PMCID: PMC11081073  PMID: 38737339

Abstract

Lead (Pb) is a nonessential heavy metal, which can cause many health problems. Isochlorogenic acid A (ICAA), a phenolic acid present in tea, fruits, vegetables, coffee, plant-based food products, and various medicinal plants, exerts multiple effects, including anti-oxidant, antiviral, anti-inflammatory and antifibrotic functions. Thus, the purpose of our study was to determine if ICAA could prevent Pb-induced hepatotoxicity in ICR mice. An evaluation was performed on oxidative stress, inflammation and fibrosis, and related signaling. The results indicate that ICAA attenuates Pb-induced abnormal liver function. ICAA reduced liver fibrosis, inflammation and oxidative stress caused by Pb. ICAA abated Pb-induced fibrosis and decreased inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-alpha (TNF-α). ICAA abrogated reductions in activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx). Masson staining revealed that ICAA reduced collagen fiber deposition in Pb-induced fibrotic livers. Western blot and immunohistochemistry analyses showed ICAA increased phosphorylated AMP-activated protein kinase (p-AMPK) expression. ICAA also reduced the expression of collagen I, α-smooth muscle actin (α-SMA), phosphorylated extracellular signal-regulated kinase (p-ERK), phosphorylated c-jun N-terminal kinase (p-JNK), p-p38, phosphorylated signal transducer and phosphorylated activator of transcription 3 (p-STAT3), transforming growth factor β1 (TGF-β1), and p-Smad2/3 in livers of mice. Overall, ICAA ameliorates Pb-induced hepatitis and fibrosis by inhibiting the AMPK/MAPKs/NF-κB and STAT3/TGF-β1/Smad2/3 pathways.

Keywords: Lead, Liver fibrosis, Isochlorogenic acid a, Oxidative stress, Inflammation, AMPK

Introduction

Lead (Pb) is a naturally occurring heavy metal that can be found in foods, drinking water, air, tobacco products; secondhand smoke, herbal remedies, electronics, toys, cosmetics and other consumer products.1,2 It is known that Pb exposure induces direct damage to several tissue and follows various pathophysiological states, including oxidative stress, inflammation and cell death.1,3,4 According to our prior research, Pb exposure can induce oxidative stress, apoptosis, and elevate the susceptibility to various forms of neurological dysfunction by the AMP-activated protein kinase (AMPK) pathway.5 Recent evidence demonstrated that Pb-induced hepatitis is possibly associated with the mitogen-activated protein kinases (MAPK) pathway.6 The previous study demonstrates that Pb caused behavioral and immune abnormalities through the activation of nuclear factor κB (NF-κB) and signal transducer and phosphorylated activator of transcription 3 (STAT3).7 Pb induced the cytotoxicity in rat liver cell lines through the upregulation of α-smooth muscle actin (α-SMA) expression in response to transforming growth factor (TGF).8 Moreover, Pb exposure caused fibrotic changes in liver, lung, testis, kidney, heart and increased collagen deposition.1,2,9–11 Nevertheless, the underlying mechanisms of liver fibrosis associated with Pb exposure are unknown.

Isochlorogenic acid A (ICAA, C25H24O12, 3,5-dicaffeoyl quinic acid), a phenolic acid present in tea, fruits, vegetables, coffee, plant-based food products, and various medicinal plants, exerts multiple effects, including anti-oxidant, antiviral, anti-inflammatory and antifibrotic properties.12–14 ICAA can affect activities of drug-metabolizing enzyme P450 and UGT in vitro and in vivo.13 ICAA mitigated LPS-induced lung injury via NF-κB/NLRP3 pathway.15 ICAA alleviated liver fibrosis and inflammation by inhibiting the high-mobility group box-1 (HMGB1)/toll-like receptor 4 (TLR4)/NF-κB pathway.12 ICAA also ameliorated experimental ulcerative colitis by inhibiting the HMGB1/TLR4/NF-κB pathway.14 We investigated whether ICAA protects liver fibrosis and clarified its mechanism in the present study.

Currently, we innovatively examined the impact of ICAA on Pb-induced oxidative stress, inflammation, and fibrosis in liver tissues. Additionally, the investigation of the AMPK/MAPKs/NF-κB and STAT3/TGF-β1/Smad2/3 signaling pathways was conducted to elucidate the underlying molecular mechanism responsible for the protective effects of ICAA.

Materials and methods

Chemicals and reagents

Isochlorogenic acid A (>98%) was obtained from Nanjing Jiangzhu Bio-Technology Co., Ltd (Nanjing, China). AMPK, p-AMPK, STAT3, p-STAT3, TGF-β1, Smad3, α-SMA, collagen I, JNK1/2, p-JNK1/2, ERK1/2, p-ERK1/2, p38, p-p38, NF-κB p65, IL-1β, TNF-α and β-actin antibodies were provided by Abcam (Cambridge, MA, USA).

Animals and ethics

Fifty male 8-week-old ICR (Institute of Cancer Research) mice were provided from Jinan experimental animal Co., Ltd, (Jiannan, China). Ethical approval was obtained from the Ethical Review Committee on Laboratory Animal Research of Jiangsu Normal University (approval No. A2023b-011).

Experimental design

Male ICR mice were provided with an ample water supply, an adequate amount of food, and were subjected to a 12-hour light/dark cycle in strict adherence to the Laboratory Animals Guidelines. Adaptive rearing for one week was followed by random assignment of mice into five different groups (10 mice/group): (1) Normal control group; (2) Pb group; (3) Pb + ICAA (20 mg/kg) group; (4) Pb + ICAA (40 mg/kg) group: (5) ICAA (40 mg/kg) control group. In groups (2), (3) and (4), the procedure of Pb-induced liver fibrosis (1 g/L lead acetate solution in drinking water for 3 months) was performed as described.11,16,17 The dosage selection for inducing fibrosis is based on previous studies.1,11 In groups (3), (4) and (5), mice were also supplied with ICAA 20 or 40 mg/kg, intragastrically once daily.12

Biochemical analysis

Liver damage and oxidative stress indexes were detected by using commercial kits (Nanjing Jiancheng, China, C010–3-1 AST assay kit, C010–3-1 ALT assay kit, C009–3-1, A003–1 MDA assay kit, A001–3 SOD assay kit, A007–1-1 CAT assay kit, A005–1-2 GPx assay kit).18–20

Histology analysis

Liver tissues were collected for fixation in 4% paraformaldehyde, followed by dehydration in increasing concentrations of ethanol and permeabilization in xylene, before being conventionally embedded in paraffin. Then, the sections were stained with hematoxylin & eosin (H&E) kits (#D006–1-1) and Masson stain kits (#D026–1-1) (Nanjing Jiangcheng). Histopathological analysis was performed to evaluate the liver damage according to previous method.18,21 The liver fibrosis was analyzed by Image J 1.42 software (NIH Bethesda, USA).

Immunohistochemistry (IHC)

Immunohistochemistry was conducted according to established protocols. Liver tissues were fixed in 4% paraformaldehyde and sectioned to a thickness of 5 μm. Following dewaxing in xylene, brain paraffin samples were rehydrated with ethanol. Microwave antigen retrieval was employed for slide treatment, and the sections were blocked with 5% BSA. Subsequently, the sections were exposed to diluted primary antibodies (Cell Signaling Technology, MA, USA) and secondary antibodies (Boster, Wuhan, China). 3,3’-Diaminobenzidine (DAB) was used to color the staining. Images of immunostained liver tissues were analyzed by Image J 1.42 software (NIH Bethesda, USA).22

Western blot analysis

The protein expression levels were analyzed by Western blot according to the manufacturer’s guidelines (Bio/Rad, Hercules, CA, USA). The blots were analyzed by Image J 1.42 software (NIH Bethesda, USA).18

Statistical analysis

Statistical analysis was performed using the ANOVA with a post hoc test. The results were shown as mean ± standard error of mean (SEM). The significance level was set at P < 0.05.

Results

ICAA reversed liver dysfunction

Comparing the Pb group with the control, serum ALT (by 232.35%) and AST (by 160.78%) activities increased statistically significantly. Conversely, a significant decrease in these serum transaminase activities was observed in the ICAA treated group in comparison to the Pb group (Table 1).

Table 1.

ICAA rescued Pb-induced liver dysfunction of mice.

ALT(U/dl) AST(U/dl)
Control 25.78 ± 0.90 a 37.12 ± 0.83 a
Pb 85.61 ± 2.67 b 96.80 ± 4.07 b
Pb + ICAA(20 mg/kg) 70.25 ± 1.19 c 80.37 ± 2.01 c
Pb + ICAA(40 mg/kg) 59.43 ± 1.95 d 66.41 ± 2.43 d
ICAA(40 mg/kg) 25.85 ± 1.12 a 37.22 ± 1.05 a
Open in a new tab

Data are expressed as mean ± standard error of mean (SEM) (n = 7). One-way ANOVA was used for comparisons of multiple group means followed by post hoc testing. Values not sharing a common superscript letter (a–d) differ significantly at P < 0.05.

ICAA alleviated liver histopathological alterations

As observed in the H&E-stained sections, a high incidence of hepatocellular necrosis and mononuclear cell infiltration was found in the livers of the Pb group. Conversely, administration of ICAA effectively mitigated the histopathological alterations in the liver. Analysis of Masson’s trichrome stained sections revealed minimal collagen fiber deposition in the livers of the control, whereas the Pb group exhibited extensive collagen fiber accumulation in liver tissue. However, treatment with ICAA resulted in a limited presence of collagen fibers in the liver tissue. The Western blot analysis demonstrated that Pb exposure resulted in an increase in the expression levels of collagen and α-SMA, when compared to the control group. Conversely, the use of ICAA exhibited a significant protective effect against liver fibrosis by inhibiting the up-regulation of collagen and α-SMA expression (Fig. 1).

Fig. 1.

Fig. 1

Open in a new tab

Isochlorogenic acid a (ICAA) improved histological alterations and fibrosis in the livers of mice. (A) Representative histological images of hematoxylin and eosin (H&E) in liver sections (×200); (B) representative histological images of Masson’s trichrome staining; (C) relative density analysis of the collagen I protein bands; (D) relative density analysis of the a-SMA protein bands. The white arrow indicates infiltrating leukocytes. The black arrow indicates collagen deposition. GAPDH was probed as an internal control in relative density analysis. The vehicle control is set as 1.0. Data are expressed as mean ± standard error of mean (SEM). (n = 3). Values not sharing a common superscript letter (a–d) differ significantly at P < 0.05.

ICAA rescued oxidative stress in livers

According to the data presented in Table 2, the Pb group exhibited a noteworthy increase in the concentration of MDA (by 110.71%) in comparison to the control. Conversely, the administration of ICAA demonstrated a substantial decrease in MDA levels in comparison to the Pb group. Additionally, the activities of SOD, GST, and CAT were significantly reduced (by 41.46%, 56.99%, 49.57%, respectively) in the Pb group when compared to the control. Nevertheless, the ICAA administration significantly increased the antioxidant enzyme activity compared to the Pb group (Table 2).

Table 2.

Effect of ICAA on Pb-induced oxidative stress in livers of mice.

MDA(nmol/mg protein) SOD(U/mg protein) GST(U/mg protein) CAT(U/mg protein)
Control 0.28 ± 0.02 a 335.33 ± 20.61a 60.27 ± 3.37 a 128.94 ± 5.93 a
Pb 0.59 ± 0.02 b 196.31 ± 5.95 b 25.92 ± 2.20 b 65.03 ± 6.14 b
Pb + ICAA(20 mg/kg) 0.45 ± 0.01 c 242.84 ± 8.02 c 34.92 ± 1.07 c 89.15 ± 7.76 c
Pb + ICAA(40 mg/kg) 0.36 ± 0.01 d 298.30 ± 10.24 d 55.16 ± 2.16 d 101.38 ± 5.09 d
ICAA(40 mg/kg) 0.27 ± 0.02 a 334.84 ± 8.39 a 60.35 ± 1.02 a 129.06 ± 6.42 a
Open in a new tab

Data are expressed as mean ± standard error of mean (SEM) (n = 7). One-way ANOVA was used for comparisons of multiple group means followed by post hoc testing. Values not sharing a common superscript letter (a–d) differ significantly at P < 0.05.

ICAA suppressed liver inflammation

Pb treatment resulted in an elevation in the levels of inflammatory mediators, such as TNF-α, IL-1β, and NF-κB p65 nuclear translocation, with increases of 732%, 1,031%, and 1,336% respectively, compared to the control. Conversely, ICAA treatment mitigated the Pb-induced rise in inflammatory factors (Fig. 2).

Fig. 2.

Fig. 2

Open in a new tab

Isochlorogenic acid a (ICAA) inhibited inflammation in the livers of mice. (A) Western blot analysis of the NF-κB p65, TNF-α and IL-1β proteins in the livers; (B) relative density analysis of the NF-κB p65 nuclear translocation; (C) relative density analysis of the TNF-α protein bands; (D) relative density analysis of the IL-1β protein bands. GAPDH and histone were probed as an internal control in relative density analysis. he vehicle control is set as 1.0. Data are expressed as mean ± standard error of mean (SEM) (n = 3). Values not sharing a common superscript letter (a–d) differ significantly at P < 0.05.

Effect of ICAA on the AMPK/MAPKs pathway

The Pb group demonstrated a notable decrease of 78% in AMPK phosphorylation levels. Conversely, the administration of ICAA (20, 40 mg/kg) resulted in an increase of 68% and 232%, respectively, in AMPK phosphorylation levels compared to the Pb group. Further confirmation was achieved by immunohistochemistry staining (Fig. 3A). In the livers of the ICAA-treated groups, the IHC signals of p-AMPK were stronger and a significantly greater amount of positive protein was detected compared to the livers of the Pb group. Additionally, Pb administration led to a significant increase in the phosphorylation of JNK1/2, ERK1/2, and p38 by 773%, 606%, and 985%, respectively, in comparison to the control. However, the administration of ICAA reduced the MAPKs phosphorylation in comparison to the Pb group (Fig. 3).

Fig. 3.

Fig. 3

Open in a new tab

Isochlorogenic acid a (ICAA) inhibited the AMPK/MAPKs pathway in the livers of mice. (A) Immunohistochemical staining of liver for p-AMPK. The scale bar represents 20 μm; (B) western blotting analysis of the AMPK/MAPKs pathway; (C) relative density analysis of the p-AMPK bands; (D) relative density analysis of the p-p38 protein bands. (E) Relative density analysis of the p-JNK1/2 bands; (F) relative density analysis of the p-ERK1/2 protein bands. AMPK, JNK1/2, ERK1/2, p38 and GAPGH were probed as the internal control in relative density analysis. The vehicle control is set as 1.0. Data are expressed as mean ± standard error of mean (SEM) (n = 3). Values not sharing a common superscript letter (a–d) differ significantly at P < 0.05.

ICAA suppressed STAT3/TGF-β1/Smad3 signaling pathway in livers

Exposure to Pb resulted in the upregulation of STAT3, leading to a significant decrease of 1,021% in STAT3 expression in comparison to the control. The administration of ICAA (20, 40 mg/kg) resulted in a reduction of STAT3 expression by 33% and 71%, respectively. Additionally, Pb exposure increased the expression of TGF-β1, a downstream factor of STAT3, by 1,082%. The administration of ICAA (20, 40 mg/kg) decreased TGF-β1 expression by 39% and 59%, respectively. Furthermore, Pb exposure increased the expression of Smad2 by 858%. The administration of ICAA (20, 40 mg/kg) reduced Smad2 expression by 17% and 36%, respectively. Pb exposure increased the expression of Smad3 by 878%. The administration of ICAA (20, 40 mg/kg) reduced Smad3 expression by 45% and 65%, respectively. These findings suggest that ICAA activates the STAT3/TGF-β1/Smad3 signaling pathway (Fig. 4).

Fig. 4.

Fig. 4

Open in a new tab

Isochlorogenic acid a (ICAA) inhibited the STAT3/TGF-β1/Smad2/3 pathway in the livers of mice. (A) Relative density analysis of the STAT3 bands;; (B) relative density analysis of the TGF-β1 bands; (C) relative density analysis of the Smad2 protein bands; (D) relative density analysis of the Smad3 bands. GAPDH was probed as the internal control in relative density analysis. The vehicle control is set as 1.0. Data are expressed as mean ± standard error of mean (SEM) (n = 3). Values not sharing a common superscript letter (a–d) differ significantly at P < 0.05.

Discussion

As a result of excessive extracellular matrix (ECM) accumulation within the liver, liver fibrosis represents a severe hepatic disease, closely related to oxidative stress and inflammation.12,16 This study provides evidence for the hepatoprotective properties of ICAA in a mouse model of liver fibrosis induced by Pb. ICAA demonstrated a significant improvement in liver functions, as evidenced by the attenuation of serum AST and ALT levels (Table 1). These beneficial effects were due to a reduction in oxidative stress and inflammation.

The present study further elucidated the impact of Pb on liver tissue by revealing a range of histological and microstructural changes, such as hepatocellular necrosis and infiltration of mononuclear cells. These findings align with our previous research outcomes.18 Additionally, numerous studies have reported that Pb induces liver fibrosis, accompanied by heightened collagen deposition and extracellular matrix production.8,9,16 Pb could also cause fibrotic changes in lung, heart, testis and kidney.2,3,9,10 The findings of the present study demonstrate that Pb exposure leads to an upregulation of fibroblast activation and subsequent overexpression of α-SMA and collagen I, suggesting an increase in ECM production and liver fibrosis (Fig. 1). Previous research has indicated that supplementation with ICAA inhibit liver fibrogenesis.12 However, our study reveals that treatment with ICAA effectively reduces the extent of fibrosis and downregulates the expression of α-SMA and collagen I in liver tissue, suggesting its potential as a protective agent against liver damage.

Several evidence suggested that inflammation played a pivotal role in the pathogenesis of hepatic fibrosis.12,15,16 Therefore, inflammatory cytokines were also detected to evaluate the anti-inflammatory effects of ICAA. Suppressing NF-κB pathway has been considered a potential anti-inflammatory signaling pathway.1,12 Several researches showed that Pb induced inflammation associated with a significant increase in the activations of NF-κB, TNF-α and IL-6.1,21,23,24 Some experiments showed that ICAA could inhibit inflammation in liver, lung and colon.12,14,15 To investigate the underlying anti-inflammatory mechanisms of ICAA, proinflammatory cytokines TNF-α and IL-1β were evaluated. The results found that the levels of TNF-α and IL-1β were upregulated in the Pb group, but significantly decreased in the Pb + ICAA group. The expression of proinflammatory factors is positively regulated by NF-κB signaling pathway. In the Pb group, nuclear migration expression was increased by protein detection. However, ICAA exhibited anti-inflammation through down-regulation of NF-κB-mediated inflammatory factor expression (Fig. 2).

The synthesis and secretion of inflammatory mediators, known as potent pro-inflammatory cytokines, are closely linked to oxidative stress, a condition that can trigger and intensify inflammation, fibrosis, and liver damage.1,25 Available data indicates that exposure to Pb results in oxidative stress within the liver.16,22,23 The present study has provided evidence that exposure to Pb resulted in oxidative stress, as indicated by an elevation in MDA concentration and a decline in the activities of SOD, CAT, and GST. In contrast, ICAA exhibited antioxidant properties by reducing MAD levels and enhancing the activities of antioxidant enzymes. ICAA administration reduced MDA concentration and increased SOD activity in the lungs of mice, according to a previous study.15 Consequently, these results suggest that ICAA effectively inhibits Pb-induced oxidative stress (Table 2).

AMPK, a prominent regulatory kinase, plays a significant role in various physiological processes such as metabolism, inflammation, autophagy, cancer, and cell death.24–26 A previous research has demonstrated that Pb has the ability to activate the AMPK signaling pathway, thereby suppressing hepatic steatosis and inflammation.26 The activation of AMPK can effectively hinder the activation of inflammasomes and subsequent inflammatory responses by inhibiting either the MAPK pathway or the NF-κB pathway, consequently reducing the production of inflammatory factors and fibrogenic factors.27–30 Exposure to Pb has been observed to upregulate the phosphorylation levels of ERK, JNK, and p38, resulting in liver damage in mice.6 We found a decrease in AMPK phosphorylation and a significant increase in the phosphorylation levels of MAPKs following treatment with ICAA. These findings suggest that ICAA may offer protection against Pb-induced liver injury by modulating the AMPK/MAPKs signaling pathway.

The liver fibrosis, a prevalent pathological alteration, is caused by the excessive accumulation of extracellular matrix (ECM) components such as fibronectin and collagen. 28,31 The activation of AMPK has been found to have a suppressive impact on the expression of STAT3, a direct transcription factor. STAT3 is known for its diverse effects in mitigating oxidative stress, inflammation and fibrosis.28,32,33 STAT3 has the capability to induce activation of the TGF/Smad signaling pathway in the context of inflammation and fibrosis. The transmission of TGF-β signaling occurs through the interaction between TGF-β receptors, which promptly phosphorylate Smad proteins and subsequently activate the R-Smad proteins. This activation initiates the transcription of various TGF-β induced target genes, leading to an upregulation in ECM protein expression and consequent progression of tissue fibrosis.28,34,35 Oxidative stress has been shown to facilitate the production and release of inflammatory mediators, which are highly potent pro-inflammatory cytokines. These cytokines play a pivotal role in initiating and intensifying inflammation, and have been linked to hepatotoxicity and fibrosis through the initiation of TGF-β signaling.34–36 Previous studies have reported that Pb exposure leads to increased activation of STAT3 in nervous tissue.4 Additionally, Pb has been found to induce activation of TGF-β1 in rat livers.8,23 Study also showed that ICAA attenuated inflammation firbosis in colon by STAT3/NF-κB pathway.14 In this investigation, the exposure to Pb demonstrated a notable augmentation in the expression levels of α-SMA, collagen I, STAT3, TGF-β1, and Smad2/3 within the liver, thereby implying the involvement of the STAT3/TGF-β1/Smad2/3 pathway in the inflammation and fibrosis induced by Pb (Fig. 4). Of utmost significance, the administration of ICAA exhibited a substantial reduction in the aforementioned protein expression induced by Pb, thereby indicating the potential of ICAA to impede inflammation and fibrosis by the STAT3/TGF-β1/Smad3 pathway.

As a result, we demonstrate that ICAA exerted the effect of inhibiting liver fibrosis by suppressing oxidative stress and inflammation. The study examined the mechanisms involved in ICAA treating live fibrosis by targeting AMPK/MAPKs/NF-κB and STAT3/TGF-β1/Smad3 pathways (Fig. 5). The mechanisms behind these findings require further research.

Fig. 5.

Fig. 5

Open in a new tab

Schematic diagram showed the possible protective effects of Isochlorogenic acid a (ICAA) in Pb-induced liver injury. The → indicates activation or induction, and Inline graphic indicates inhibition or blockade.

Author contributions

All authors reviewed and approved the final manuscript. CL made contributions to the conception and design of the study. JG was responsible for designing experiments and drafting the manuscript. HL and CC conducted the experiments, while JL prepared the experimental materials. HR analyzed the experimental data, and JS reviewed and revised the final version of the manuscript.

Funding

This work is supported by the National Natural Science Foundation of China (31972942). The author thanks all those who participated in and supported this study.

Conflict of interest statement: The authors have no relevant financial or non-financial interests to disclose.

Data availability

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

Supplementary Material

Supplementary_WB_tfae072
supplementary_wb_tfae072.docx (570.5KB, docx)
Supplementary_activity_tfae072
supplementary_activity_tfae072.xlsx (25.1KB, xlsx)

Contributor Information

Jun-Tao Guo, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

Han-Yu Li, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

Chao Cheng, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

Jia-Xue Shi, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

Hai-Nan Ruan, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

Jun Li, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

Chan-Min Liu, School of Life Science, Jiangsu Normal University, No. 101, Shanghai Road, Tongshan New Area, 221116, Xuzhou City, Jiangsu Province, PR China.

References

  • 1. Alhusaini A, Fadda L, Hasan IH, Zakaria E, Alenazi AM, Mahmoud AM. Curcumin ameliorates lead-induced hepatotoxicity by suppressing oxidative stress and inflammation, and modulating Akt/GSK-3β signaling pathway. Biomol Ther. 2019:9(11):703. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Liu Q, Xu C, Jin J, Li W, Liang J, Zhou S, Weng Z, Zhou Y, Liao X, Gu A. Early-life exposure to lead changes cardiac development and compromises long-term cardiac function. Sci Total Environ. 2023:904:166667. [DOI] [PubMed] [Google Scholar]
  • 3. Shafai AE, Zohdy N, Mulla KE, Hassan M, Morad N. Light and electron microscopic study of the toxic effect of prolonged lead exposure on the seminiferous tubules of albino rats and the possible protective effect of ascorbic acid. Food Chem Toxicol. 2011:49(4):734–743. [DOI] [PubMed] [Google Scholar]
  • 4. Beier EE, Maher JR, Sheu TJ, Cory-Slechta DA, Berger AJ, Zuscik MJ, Puzas JE. Heavy metal lead exposure, osteoporotic-like phenotype in an animal model, and depression of Wnt signaling. Environ Health Perspect. 2013:121:97–104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5. Yang W, Tian ZK, Yang HX, Feng ZJ, Sun JM, Jiang J, Cheng C, Ming QL, Liu CM. Fisetin improves lead-induced neuroinflammation, apoptosis and synaptic dysfunction in mice associated with the AMPK/SIRT1 and autophagy pathway. Food Chem Toxicol. 2019:139:110824. [DOI] [PubMed] [Google Scholar]
  • 6. Qiu T, Shi JX, Cheng C, Jiang H, Ruan HN, Li J, Liu CM. Hepatoprotective effect of avicularin on lead-induced steatosis, oxidative stress, and inflammation in mice associated with the MAPK/HSP60/NLRP3 and SREBP1c pathway. Toxicol Res (Camb). 2023:12(3):417–424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. Almutairi MM, Nadeem A, Ansari MA, Bakheet SA, Attia SM, Albekairi TH, Alhosaini K, Algahtani M, Alsaad AMS, Al-Mazroua HA, et al. Lead (Pb) exposure exacerbates behavioral and immune abnormalities by upregulating Th17 and NF-κB-related signaling in BTBR T+Itpr3tf/J autistic mouse model. Neurotoxicology. 2022:91:340–348. [DOI] [PubMed] [Google Scholar]
  • 8. Hwang JY, Lin JT, Liu SC, Hu CC, Shyu YS, Yang DJ. Protective role of litchi (litchi chinensis Sonn.) flower extract against cadmium- and lead-induced cytotoxicity and transforming growth factor b1-stimulated expression of smooth muscle α-actin estimated with rat liver cell lines. J Funct Foods. 2013:5:698–705. [Google Scholar]
  • 9. Riaz MA, Nisa ZU, Mehmood A, Anjum MS, Shahzad K. Metal-induced nephrotoxicity to diabetic and non-diabetic Wistar rats. Environ Sci Pollut Res. 2019:26(30):31111–31118. [DOI] [PubMed] [Google Scholar]
  • 10. Yau WH, Chen SC, Wu DW, Chen HC, Lin HH, Wang CW, Hung CH, Kuo CH. Blood lead (Pb) is associated with lung fibrotic changes in non-smokers living in the vicinity of petrochemical complex: a population based study. Environ Sci Pollut Res. 2023:30:75225–75234. [DOI] [PubMed] [Google Scholar]
  • 11. Zheng X, Guo C, Lv Z, Jiang H, Li S, Yu L, Zhang Z. From animal to cell model: Pyroptosis targeted-fibrosis is a novel mechanism of lead-induced testicular toxicity. Food Chem Toxicol. 2023:178:113886. [DOI] [PubMed] [Google Scholar]
  • 12. Liu X, Huang K, Zhang RJ, Mei D, Zhang B. Isochlorogenic acid a attenuates the progression of liver fibrosis through regulating HMGB1/TLR4/NF-kB signaling pathway. Front Pharmacol. 2020:11:582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. Wang J, Wang H, Peng Y, Wang GJ, Hao HP. Isochlorogenic acid a affects P450 and UGT enzymes in vitro and in vivo. Chin J Nat Med. 2016:14(11):865–870. [DOI] [PubMed] [Google Scholar]
  • 14. Tang S, Zhong W, Li T, Li Y, Song G. Isochlorogenic acid a alleviates dextran sulfate sodium-induced ulcerative colitis in mice through STAT3/NF-кB pathway. Int Immunopharmacol. 2023:118:109989. [DOI] [PubMed] [Google Scholar]
  • 15. Wang Q, Xiao L. Isochlorogenic acid a attenuates acute lung injury induced by LPS via Nf-κB/NLRP3 signaling pathway. Am J Transl Res. 2019:11(11):7018–7026. [PMC free article] [PubMed] [Google Scholar]
  • 16. Soliman MM, Baiomy AA, Yassin MH. Molecular and histopathological study on the ameliorative effects of curcumin against lead acetate-induced hepatotoxicity and nephrototoxicity in Wistar rats. Biol Trace Elem Res. 2015:167:91–102. [DOI] [PubMed] [Google Scholar]
  • 17. Yang D, Jiang H, Lu J, Lv Y, Baiyun R, Li S, Liu B, Lv Z, Zhang Z. Dietary grape seed proanthocyanidin extract regulates metabolic disturbance in rat liver exposed to lead associated with PPARα signaling pathway. Environ Pollut. 2018:237:377–387. [DOI] [PubMed] [Google Scholar]
  • 18. Qiu T, Shi JX, Cheng C, Jiang H, Ruan HN, Li J, Liu CM. Avicularin attenuates lead-induced impairment of hepatic glucose metabolism by inhibiting the ER stress-mediated inflammatory pathway. Nutrients. 2022:14:4806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Yuan E, Duan X, Xiang L, Ren J, Lai X, Li Q, Sun L, Sun S. Aged oolong tea reduces high-fat diet-induced fat accumulation and dyslipidemia by regulating the AMPK/ACC signaling pathway. Nutrients. 2018:10:187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. Sun W, Lei Y, Jiang Z, Wang K, Liu H, Xu T. BPA and low-Se exacerbate apoptosis and mitophagy in chicken pancreatic cells by regulating the PTEN/PI3K/AKT/mTOR pathway. J Adv Res. 2024:S2090-1232(24):00042-0. [DOI] [PubMed] [Google Scholar]
  • 21. Miao Z, Miao Z, Liu M, Xu S. Melatonin ameliorates imidacloprid-induced intestinal injury by negatively regulating the PGN/P38MAPK pathway in the common carp (Cyprinus carpio). Fish Shellfish Immunol. 2022:131:1063–1074. [DOI] [PubMed] [Google Scholar]
  • 22. Miao Z, Miao Z, Teng X, Xu S. Melatonin alleviates lead-induced fatty liver in the common carps (Cyprinus carpio) via gut-liver axis. Environ Pollut. 2023:317:120730. [DOI] [PubMed] [Google Scholar]
  • 23. Almasmoum H, Refaat B, Ghaith MM, Almaimani RA, Sr I, Ahmad J, Abdelghany AH, BaSalamah MA, El-Boshy M. Protective effect of vitamin D3 against lead induced hepatotoxicity, oxidative stress, immunosuppressive and calcium homeostasis disorders in rat. Environ Toxicol Pharmacol. 2019:72:103246. [DOI] [PubMed] [Google Scholar]
  • 24. Chibowska K, Baranowska-Bosiacka I, Falkowska A, Gutowska I, Goschorska M, Chlubek D. Effect of lead (Pb) on inflammatory processes in the brain. Int J Mol Sci. 2016:17:2140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. Hou G, Surhio MM, Ye H, Gao X, Ye Z, Li J, Ye M. Protective effects of a Lachnum polysaccharide against liver and kidney injury induced by lead exposure in mice. Int J Biol Macromol. 2019:124:716–723. [DOI] [PubMed] [Google Scholar]
  • 26. Miao Z, Miao Z, Feng S, Xu S. Chlorpyrifos-mediated mitochondrial calcium overload induces EPC cell apoptosis via ROS/AMPK/ULK1. Fish Shellfish Immunol. 2023:141:109053. [DOI] [PubMed] [Google Scholar]
  • 27. Xu L, Sun X, Zhu G, Mao J, Baban B, Qin X. Local delivery of simvastatin maintains tooth anchorage during mechanical tooth moving via anti-inflammation property and AMPK/MAPK/NF-kB inhibition. J Cell Mol Med. 2021:25(1):333–344. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 28. Winarto J, Song DG, Pan CH. The role of fucoxanthin in non-alcoholic fatty liver disease. Int J Mol Sci. 2023:24:8203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 29. Seoane-Collazo P, Martínez de Morentin PB, Fernø J, Diéguez C, Nogueiras R, López M. Nicotine improves obesity and hepatic steatosis and ER stress in diet-induced obese male rats. Endocrinology. 2014:155(5):1679–1689. [DOI] [PubMed] [Google Scholar]
  • 30. Chen Y, Ge Z, Huang S, Zhou L, Zhai C, Chen Y, Hu Q, Cao W, Weng Y, Li Y. Delphinidin attenuates pathological cardiac hypertrophy via the AMPK/NOX/MAPK signaling pathway. Aging. 2020:12:5362–5383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31. Liang Z, Li T, Jiang S, Xu J, Di W, Yang Z, Hu W, Yang Y. AMPK: a novel target for treating hepatic fibrosis. Oncotarget. 2017:8:62780–62792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. Cheng F, Yuan G, He J, Shao Y, Zhang J, Guo X. Dysregulation of DPP4 is associated with the AMPK/JAK2/STAT3 pathway in adipocytes under insulin resistance status and liraglutide intervention. Diabetes Metab Syndr Obes. 2019:12:2635–2644. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. Xu Y, Wang N, Tan HY, Li S, Zhang C, Zhang Z, Feng Y. Panax notoginseng saponins modulate the gut microbiota to promote thermogenesis and beige adipocyte reconstruction via leptin-mediated AMPKα/STAT3 signaling in diet-induced obesity. Theranostics. 2020:10(24):11302–11132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. Akool ES, Doller A, Babelova A, Tsalastra W, Moreth K, Schaefer L, Pfeilschifter J, Eberhardt W. Molecular mechanisms of TGFβ receptor-triggered signaling cascades rapidly induced by the calcineurin inhibitors cyclosporin a and FK5061. J Immunol. 2008:181:2831–2845. [DOI] [PubMed] [Google Scholar]
  • 35. Wu M, Xu H, Liu J, Tan X, Wan S, Guo M, Long Y, Xu Y. Metformin and fibrosis: a review of existing evidence and mechanisms. J Diabetes Res. 2021:2021:1–11. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36. Shi Y, Shi L, Liu Q, Wang W, Liu Y. Molecular mechanism and research progress on pharmacology of ferulic acid in liver diseases. Front Pharmacol. 2023:14:1207999. [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.

Supplementary Materials

Supplementary_WB_tfae072
supplementary_wb_tfae072.docx (570.5KB, docx)
Supplementary_activity_tfae072
supplementary_activity_tfae072.xlsx (25.1KB, xlsx)

Data Availability Statement

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

Articles from Toxicology Research are provided here courtesy of Oxford University Press

RESOURCES