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. 2023 Nov 15;28(22):7608.
doi: 10.3390/molecules28227608.

Hepatoprotective Effects of Biochanin A on Thioacetamide-Induced Liver Cirrhosis in Experimental Rats

Mohamed Yousif Ibrahim  1 , Zaenah Zuhair Alamri  2 , Ameena S M Juma  3 , Sarah Ashour Hamood  4 , Suhayla Hamad Shareef  5 , Mahmood Ameen Abdulla  3 , Soher Nagi Jayash  6
Affiliations

Hepatoprotective Effects of Biochanin A on Thioacetamide-Induced Liver Cirrhosis in Experimental Rats

Mohamed Yousif Ibrahim et al. Molecules. .

Abstract

The protective effect of biochanin A (BCA) on the histopathology, immunohistochemistry, and biochemistry of thioacetamide (TAA)-induced liver cirrhosis in vivo was investigated. There was a significant reduction in liver weight and hepatocyte propagation, with much lower cell injury in rat groups treated with BCA (25 mg/kg and 50 mg/kg) following a TAA induction. These groups had significantly lower levels of proliferating cell nuclear antigen (PCNA) and α-smooth muscle actin (α-SMA). The liver homogenates showed increased antioxidant enzyme activity of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), as well as decreased malondialdehyde (MDA) levels. The serum biomarkers associated with liver function, namely alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma glutamyl transaminase (GGT), returned to normal levels, comparable to those observed in both the normal control group and the reference control group. Taken together, the normal microanatomy of hepatocytes, the inhibition of PCNA and α-SMA, improved antioxidant enzymes (SOD, CAT, and GPx), and condensed MDA with repairs of liver biomarkers validated BCA's hepatoprotective effect.

Keywords: TAA; antioxidant enzymes; biochanin A; histopathology; immunohistochemistry; liver cirrhosis.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Chemical Structure of Biochanin A.
Figure 2
Figure 2
Effect of biochanin A (BCA) on histological sections of the liver (1st column) and kidney (2nd column) from acute toxicity test in rats. (A,B) Normal control group (C,D) rats fed with 20 mg g/kg of BCA (E,F) rats fed with 200 mg/kg of BCA. There is no significant change in the architecture of the livers and kidneys between the treated and control groups (Hematoxylin and Eosin stain, 20×). Scale bar 100 µm.
Figure 3
Figure 3
Effects of BCA on thioacetamide-induced liver injury in experimental rats. (A) Normal control group, (B) rats treated with thioacetamide (TTA group), (C) rats treated with thioacetamide + 50 mg/kg silymarin (silymarin group), (D) rats treated with thioacetamide + 25 mg/kg BCA (BCA low dose group), and (E) Rats treated with thioacetamide + 50 mg/kg BCA (BCA high dose group). (GA) Gross morphology, Hematoxylin and Eosin (H&E) stain 20×, (MT) Masson trichrome stain 20×.
Figure 4
Figure 4
Immunohistochemistry staining for (PCNA) of liver sections sampled from different groups. (A) Normal control group, no PCNA staining (downregulation); (B) rats treated with thioacetamide (TTA group)), PCNA expression in hepatocyte nuclei (upregulation); (C) rats treated with thioacetamide + 50 mg/kg silymarin (silymarin group), mild PCNA expression in hepatocytes nuclei (downregulation); (D) rats treated with thioacetamide + 25 mg/kg BCA (BCA low-dose group), mild to moderate expression in PCNA-positive hepatocyte nuclei (downregulation); and (E) rats treated with thioacetamide + 50 mg/kg BCA (BCA high-dose group), mild PCNA expression in hepatocyte nuclei (downregulation) (original magnification ×20). Scale bar 50 µm.
Figure 5
Figure 5
Immunohistochemistry staining of alpha smooth muscle actin (α-SMA) of liver sections samples from the different groups. (A) The livers from the normal control group showed no signs of positive α-SMA staining. (B) The liver from the TAA group showed dense brown staining with highly expressed α-SMA, demonstrating highly activated hepatic stellate cells. (C) Very little positive α-SMA staining in the silymarin-treated liver from the silymarin-treated group. (D) Moderate positive staining of α-SMA in the liver from BCA 25 mg/kg Group. (E) Very few positive α-SMA staining reflecting very few activated hepatic stellate cells in the liver from BCA 50 mg/kg of Group.
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