Silymarin and N-acetylcysteine reduce acetaminophen-induced liver damage in rats by modulating the expression of CYP2E1 and Nrf2 genes

Document Type : Research Paper

Authors

1 Department of Medical Sciences, QoMS.C, Islamic Azad University, Qom, Iran

2 Department of Basic Medical Sciences, Termez University of Economics and Service, Termez, Uzbekistan

3 Department of Biology, Urgench State University, Urgench, Uzbekistan

4 Department of Natural Sciences, Candidate of Medical Sciences, Mamun University, Uzbekistan

5 Department of Clinical and Laboratory Diagnostics, Samarkand State Medical University. Samarkand, Uzbekistan

6 Biology Teacher of the Department of "Biology", Urgench State University, Uzbekistan

7 Department of Clinical Sciences, Ma'mun University, Urgench, Uzbekistan

8 Department of Primary Education Methodology, Termiz University of Economics and Service, Termez, Uzbekistan

9 Department of English, College of Science and Humanities, Al-Kharj, Prince Sattam Bin Abdulaziz University, Al-Kharj, 11942, Saudi Arabia

10 Department of Biology, Ard.C., Islamic Azad University, Ardebil, Iran

10.22124/cjes.2025.9293

Abstract

While silymarin and N-acetylcysteine have well-established individual protective properties against acetaminophen (APAP)-induced liver injury, their synergistic effects on key genes, such as CYP2E1 and Nrf2, have not been thoroughly investigated in a combined prophylactic and early post-exposure regimen. In a rat model of APAP-induced acute liver injury, this study examined the hepatoprotective effects of silymarin and NAC, both alone and in combination, with a focus on controlling CYP2E1 and Nrf2 mRNA expression. Thirty male Wistar rats were divided into five groups: the APAP-only group, the APAP+silymarin group, the APAP+NAC group, and the APAP+silymarin+NAC group. Hepatic damage was caused by a single peritoneal administration of 750 mg/kg of APAP. Rats were given NAC (300 mg/kg) or silymarin (100 mg/kg) orally two hours before and four hours after the APAP injection. The activity of antioxidant enzymes (SOD, CAT, and GPx), the levels of ALT, AST, and ALP in the serum, the amounts of MDA and GSH in the liver, and the mRNA levels of Nrf2 and CYP2E1 were all measured using qPCR. Liver slices were examined under a microscope using HandE staining. Statistical comparisons were carried out using one-way ANOVA, followed by Tukey’s post-hoc test. When APAP was administered, blood liver enzymes, MDA levels, and histological necrosis increased significantly, while GSH and antioxidant enzyme activity decreased significantly (p<0.001). The most notable improvements came from the combination of silymarin and NAC treatment: the fastest return to normalcy of liver enzymes, the largest decrease in MDA levels, the strongest recovery of GSH levels, the most obvious downregulation of CYP2E1, and the most obvious upregulation of Nrf2 (all p<0.001 compared to either drug alone). Little protection was provided by monotherapy. Histological necrosis scores were highest in the APAP-only group (3.5 ± 0.5) and lowest in the combined group (0.75 ± 0.25). This suggests a marked decrease in both cellular apoptosis and inflammation. In conclusion, compared to using either agent alone, concurrent administration of silymarin and NAC in a preventive and early post-exposure protocol offers superior hepatoprotection primarily through the beneficial modulation of CYP2E1 and Nrf2 expression. According to these findings, this combination might be used as an additional treatment in preclinical models of acute liver injury caused by APAP.

Keywords

References

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Caspian Journal of Environmental Sciences
Volume 23, Issue 5
Special issue: Biological Researches and Envireonment, Guest Editor: Prof. Hamed Mousavi-Sabet, University of Guilan, Iran
December 2025
Pages 1151-1158

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