Aqueous extract of Amydrium sinense (Engl.) H. Li alleviates hepatic fibrosis by suppressing hepatic stellate cell activation through inhibiting Stat3 signaling

Jingyan Li; Bingmin Wu; Lishan Zeng; Ying Lin; Qiuhe Chen; Haixia Wang; Lin An; Jiajun Zhang; Siyan Chen; Junying Huang; Ruoting Zhan; Guifang Zhang

doi:10.3389/fphar.2023.1101703

Aqueous extract of Amydrium sinense (Engl.) H. Li alleviates hepatic fibrosis by suppressing hepatic stellate cell activation through inhibiting Stat3 signaling

Front Pharmacol. 2023 Jun 13:14:1101703. doi: 10.3389/fphar.2023.1101703. eCollection 2023.

Authors

Jingyan Li¹, Bingmin Wu¹, Lishan Zeng¹, Ying Lin¹, Qiuhe Chen¹, Haixia Wang¹, Lin An¹, Jiajun Zhang¹, Siyan Chen¹, Junying Huang², Ruoting Zhan^{1

3}, Guifang Zhang^{1

3}

Affiliations

¹ Guangdong Key Laboratory for Translational Cancer Research of Chinese Medicine, Joint Laboratory for Translational Cancer Research of Chinese Medicine of the Ministry of Education of the People's Republic of China, International Institute for Translational Chinese Medicine, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong, China.
² College of Life Sciences, Guangzhou University, Guangzhou, Guangdong, China.
³ Key Laboratory of Chinese Medicinal Resource from Lingnan, Ministry of Education, Guangzhou University of Chinese Medicine, Guangzhou, China.

Abstract

Background: The present study aimed to investigate the protective effect of the water extract of Amydrium sinense (Engl.) H. Li (ASWE) against hepatic fibrosis (HF) and clarify the underlying mechanism. Methods: The chemical components of ASWE were analysed by a Q-Orbitrap high-resolution mass spectrometer. In our study, an in vivo hepatic fibrosis mouse model was established via an intraperitoneal injection of olive oil containing 20% CCl₄. In vitro experiments were conducted using a hepatic stellate cell line (HSC-T6) and RAW 264.7 cell line. A CCK-8 assay was performed to assess the cell viability of HSC-T6 and RAW264.7 cells treated with ASWE. Immunofluorescence staining was used to examine the intracellular localization of signal transducer and activator of transcription 3 (Stat3). Stat3 was overexpressed to analyse the role of Stat3 in the effect of ASWE on HF. Results: Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses showed that candidate targets of ASWE, associated with protective effects against hepatic fibrosis, were related to inflammation response. ASWE ameliorated CCl₄-induced liver pathological damage and reduced the liver index and alanine transaminase (ALT) and aspartate transaminase (AST) levels. ASWE also decreased the serum levels of collagen Ⅰ (Col Ⅰ) and hydroxyproline (Hyp) in CCl₄-treated mice. In addition, the expression of fibrosis markers, including α-SMA protein and Acta2, Col1a1, and Col3a1 mRNA, was downregulated by ASWE treatment in vivo. The expression of these fibrosis markers was also decreased by treatment with ASWE in HSC-T6 cells. Moreover, ASWE decreased the expression of inflammatory markers, including the Tnf-α, Il6 and Il1β, in RAW264.7 cells. ASWE decreased the phosphorylation of Stat3 and total Stat3 expression and reduced the mRNA expression of the Stat3 gene in vivo and in vitro. ASWE also inhibited the nuclear shuttling of Stat3. Overexpression of Stat3 weakened the therapeutic effect of ASWE and accelerated the progression of HF. Conclusion: The results show that ASWE protects against CCl₄-induced liver injury by suppressing fibrosis, inflammation, HSC activation and the Stat3 signaling pathway, which might lead to a new approach for preventing HF.

Keywords: Amydrium sinense; Stat3; carbon tetrachloride; hepatic fibrosis; α-SMA.

Grants and funding

This work was supported by grants from the National Natural Science Foundation of China (Nos 82104163 and 82003826), Guangdong Basic and Applied Basic Research Foundation (Nos 2019A1515110607 and 2021A1515011016), Guangdong Science and Technology Project “Overseas Master” Project (No. 2020A141401022), Guangzhou Basic Research Program (No. 202102010376), Project of Traditional Chinese Medicine Bureau of Guangdong Province (No. 20221117), Project of south medicine innovation team in modern agricultural industry technology system of Guangdong Province (No. 2022KJ148) and the Special Funds in Key Areas of “Serving Rural Revitalization Plan” for Higher Education Institutions of Guangdong Province (2019KZDZX 2017).