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Hepatoprotective effect and possible mechanism of phytoestrogen calycosin on carbon tetrachloride–induced liver fibrosis in mice

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Abstract

The study was to explore the hepatoprotective effect and possible mechanism of calycosin on carbon tetrachloride (CCl4)–induced liver fibrosis in mice. Hepatic fibrosis was induced by intraperitoneal injection of CCl4 in C57BL/6 male mice. Serum alanine aminotransferase (ALT) and aspartate transaminase (AST) activity, superoxide dismutase (SOD) activity, and hydroxyproline (Hyp) and malondialdehyde (MDA) levels were determined by biochemical assays. Liver histopathology was assessed by H&E and Masson trichrome staining. The mRNA expressions of α-smooth muscle actin (α-SMA), collagen-I (Col-I), Janus kinase 2 (JAK2) and signal transducer and activator of transcription 3 (STAT3) were determined using qRT-PCR. The protein levels of α-SMA, Col-I, estrogen receptor α (ERα), estrogen receptor β (ERβ), tissue inhibitor of metalloproteinase-1 (TIMP-1), matrix metalloproteinase-1 (MMP-1), JAK2, phospho-JAK2 (p-JAK2), STAT3, and phospho-STAT3 (p-STAT3) were detected by Western blotting. The levels of α-SMA and ERβ were measured by immunohistochemistry. Calycosin significantly reduced liver index, MDA level, and ALT and AST activity and increased SOD activity. The α-SMA, Col-I, and Hyp of the calycosin group were significantly lower than those of the model group. Calycosin increased MMP-1 and inhibited TIMP-1 expression resulting in the improvement of MMP-1/TIMP-1 ratio. Importantly, calycosin improved ERβ protein expression, JAK2 and STAT3 mRNA expressions, p-JAK2/JAK2, and p-STAT3/STAT3 relative protein expressions. However, ERα, JAK2, and STAT3 protein expressions were relatively unchanged. Calycosin significantly inhibits liver fibrosis in mice, and its mechanism may involve the following: calycosin inhibits oxidative stress; calycosin inhibits collagen synthesis and balances MMP-1/TIMP-1 system; calycosin increases ERβ expression and activates JAK2-STAT3 pathway.

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Acknowledgments

The drug identification (Fig. 2) was from Professor Jingbo Shi (School of Pharmacy, Anhui Medical University, Anhui, China).

Funding

This work was supported by the University Natural Science Research Project of Anhui Province (NO. KJ2016A342).

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Authors and Affiliations

  1. Department of Pharmacology, School of Basic Medical Sciences, Anhui Medical University, Hefei, 230032, Anhui Province, China

    Mengmeng Zhang, Yaxin Wang, Guannan Zhu, Cheng Sun & Jiajia Wang

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  1. Mengmeng Zhang
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Contributions

Mengmeng Zhang and Jiajia Wang designed research. Mengmeng Zhang and Yaxin Wang conducted experiments. Mengmeng Zhang and Guannan Zhu analyzed data. Mengmeng Zhang and Cheng Sun wrote the manuscript. All authors read and approved the manuscript. All experimental data were generated in-house and we did not use a paper mill.

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Correspondence to Jiajia Wang.

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In our study, all animal experiments were reviewed and approved by the Ethical Committee and the China National Institutes of Healthy Guidelines for the Care and Use of Laboratory Animals.

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Zhang, M., Wang, Y., Zhu, G. et al. Hepatoprotective effect and possible mechanism of phytoestrogen calycosin on carbon tetrachloride–induced liver fibrosis in mice. Naunyn-Schmiedeberg's Arch Pharmacol 394, 189–204 (2021). https://doi.org/10.1007/s00210-020-01891-5

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