Huagan tongluo Fang improves liver fibrosis via down-regulating miR-184 and up-regulating FOXO1 to inhibit Th17 cell differentiation

Ji Xuan; Ang Huang; Dashan Hu; Jiabao Geng; Yaozhou Tian; Zhengyuan Cheng; Yuping Qiu

doi:10.1016/j.yexmp.2020.104447

Huagan tongluo Fang improves liver fibrosis via down-regulating miR-184 and up-regulating FOXO1 to inhibit Th17 cell differentiation

Exp Mol Pathol. 2020 Aug:115:104447. doi: 10.1016/j.yexmp.2020.104447. Epub 2020 May 4.

Authors

Ji Xuan¹, Ang Huang², Dashan Hu³, Jiabao Geng⁴, Yaozhou Tian⁵, Zhengyuan Cheng¹, Yuping Qiu¹

Affiliations

¹ Department of Gastroenterology, Jinling Hospital, Nanjing 210002, Jiangsu, China.
² Department of non-infection liver disease, The Center of Liver Disease, The Fifth Medical Center of Chinese PLA General Hospital, Beijing 100039, China.
³ Department of infection internal medicine, The Eighth Second Hospital of the General Hospital of the East War Zone, Huaian 223001, Jiangsu, China.
⁴ Department of infection internal medicine, Jinling Hospital, Nanjing 210002, Jiangsu, China.
⁵ Department of Gastroenterology, Jiangsu Branch of China Academy of Chinese Medical Sciences, Nanjing 210002, Jiangsu, China. Electronic address: tianyaozhou1960@163.com.

PMID: 32380055
DOI: 10.1016/j.yexmp.2020.104447

Abstract

Background: The purpose of this research is to reveal the improvement effect and potential mechanism of Huagan tongluo Fang (HGTLF) on liver fibrosis.

Methods: A mouse model of liver fibrosis induced by CCl₄ was established to analyze the effect of HGTLF on liver fibrosis. The expression changes of miRNA after HGTLF stimulation were detected by qRT-PCR. After interference with miR-184 in Th17 cells, the concentration of IL-17A in cell culture supernatants was detected by ELISA and the proportion of Th17 cells was analyzed by flow cytometry. The relationship between miR-184 and FOXO1 was verified by online software and dual-luciferase reporter system. After HGTLF treatment of Th17 cells overexpressing miR-184, the protein level of FOXO1 was detected by Western blot.

Results: HGTLF could significantly improve liver fibrosis in mice. By qRT-PCR, miR-184 was most significantly expressed after HGTLF drug stimulation, and miR-184 was considered to be the major RNA involved in Th17 cell differentiation. Interference with miR-184 in Th17 cells inhibited the differentiation of Th17 cells. By online software and dual-luciferase reporter system assay, the direct interaction of miR-184 with FOXO1 was confirmed. After HGTLF treatment of Th17 cells overexpressing miR-184, FOXO1 protein levels were significantly up-regulated and inhibited the differentiation of Th17 cells, which was reversed by miR-184 inhibitors. The Vivo experiments also confirmed the improvement effect of HGTLF on liver fibrosis in mice.

Conclusion: Our results indicated that HGTLF could improve liver fibrosis via down-regulating miR-184 and up-regulating of FOXO1 to inhibit Th17 cell differentiation.

Keywords: FOXO1; HGTLF; Liver fibrosis; Th17 cell; miR-184.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Base Sequence
Carbon Tetrachloride
Cell Differentiation*
Disease Models, Animal
Down-Regulation / genetics*
Drugs, Chinese Herbal / pharmacology
Drugs, Chinese Herbal / therapeutic use*
Forkhead Box Protein O1 / genetics
Forkhead Box Protein O1 / metabolism*
Liver Cirrhosis / drug therapy*
Liver Cirrhosis / genetics
Liver Cirrhosis / immunology
Male
Mice, Inbred C57BL
MicroRNAs / genetics*
MicroRNAs / metabolism
Th17 Cells / cytology*
Up-Regulation / genetics*

Substances

Drugs, Chinese Herbal
FOXO1 protein, human
Forkhead Box Protein O1
MIRN184 microRNA, human
MicroRNAs
tongluo
Carbon Tetrachloride