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栀子苷通过TGF-β1/Smad信号通路抑制肝纤维化和肝星状细胞活化

周林华^1,*, 陈晓²

¹宜春学院美容医学院，宜春 336000；²宜春职业技术学院护理学院，宜春 336000

摘要

本研究旨在观察栀子苷对肝纤维化和肝星状细胞活化的影响，并探讨其可能机制。人肝星状细胞LX-2用5 ng/mL转化生长因子-β1 (transforming growth factor-β1, TGF-β1)处理，并用不同浓度(0, 1, 2.5, 5, 10, 20, 40, 60, 80, 100 μmol/L)的栀子苷联合培养，MTT法进行细胞活力测定，LX-2分别经5 ng/mL TGF-β1、TGF-β1+栀子苷(20 μmol/L)处理后，用qPCR和Western blot分别检测I型胶原蛋白(collagen I)、纤连蛋白(fibronectin)、α平滑肌肌动蛋白(α-smooth muscle actin, α-SMA)、p-Smad2和p-Smad3基因和蛋白的表达。BALB/c小鼠用CCl4 (25%, 1 mL/kg)诱导肝纤维化，设立对照组、CCl4组和CCl4+栀子苷组(40 mg/kg)，4周后检测肝功能及血清肝纤维化指标，用HE和Masson染色进行组织学观察，免疫组织化学分析组织α-SMA表达，Western blot检测α-SMA、TGF-β1、p-Smad2和p-Smad3蛋白的表达。结果显示，栀子苷显著抑制TGF-β1诱导的LX-2细胞增殖、collagen I、fibronectin和α-SMA的基因和蛋白表达及TGF-β1/Smad信号相关蛋白表达。组织学观察显示栀子苷显著抑制CCl4诱导的小鼠肝纤维化，显著抑制肝星状细胞活化和TGF-β1/Smad信号相关蛋白表达。以上结果提示，栀子苷抑制肝纤维化和肝星状细胞活化，其机制可能是通过抑制TGF-β1/Smad信号传导。

关键词： 栀子苷; 肝纤维化; 肝星状细胞; TGF-β1/Smad

Geniposide inhibits hepatic fibrosis and hepatic stellate cell activation through blocking the TGF-β1/Smad signaling pathway

ZHOU Lin-Hua^1,*, CHEN Xiao²

¹School of Cosmetology, Yichun University, Yichun 336000, China；²School of Nursing, Yichun Vocational Technical College, Yichun 336000, China

Abstract

The purpose of this study was to investigate the effect of Geniposide on hepatic fibrosis and activation of hepatic stellate cells (HSCs) and to explore possible underlying mechanism. Human HSCs (LX-2) were treated with 5 ng/mL transforming growth factor-β1 (TGF-β1), followed by co-culture with Geniposide at various concentrations (0, 1, 2.5, 5, 10, 20, 40, 60, 80, 100 μmol/L). Cell viability was determined by MTT assay. Then, LX-2 cells were divided into control, TGF-β1 (5 ng/mL) and TGF-β1 + Geniposide (20 μmol/L) groups, and the gene and protein expression of collagen I, fibronectin, α-smooth muscle actin (α-SMA), p-Smad2 and p-Smad3 was detected by qPCR and Western blot, respectively. BALB/c mice were treated with CCl4 (25%, 1 mL/kg) to generate a model of hepatic fibrosis (CCl4 group), and the control group and CCl4 + Geniposide group were administered with olive oil and CCl4 + 40 mg/kg Geniposide, respectively. After 4 weeks of treatment, the liver function and serum hepatic fibrosis indexes of mice were detected, histological observation was performed by HE and Masson staining, and α-SMA expression in the tissue was analyzed by immunohistochemistry. Western blot was utilized for the determination of the protein expression of α-SMA, TGF-β1, p-Smad2 and p-Smad3. The results showed that Geniposide inhibited LX-2 cell proliferation. In addition, Geniposide significantly downregulated the gene and protein expression of collagen I, fibronectin and α-SMA and the expression of TGF-β1/Smad signaling-related proteins induced by TGF-β1 in vitro. Histological observations showed that Geniposide significantly inhibited CCl4-induced hepatic fibrosis, HSC activation and expression of TGF-β1/Smad signaling-related proteins in mice. In summary, Geniposide prevents the hepatic fibrosis and HSC activation possibly through the inhibition of the TGF-β1/Smad signaling pathway.

Key words: Geniposide; hepatic fibrosis; hepatic stellate cell; TGF-β1/Smad

收稿日期：　　录用日期：

通讯作者：周林华　　E-mail: ycxyzlh@163.com

DOI: 10.13294/j.aps.2022.0019

引用本文：

周林华, 陈晓. 栀子苷通过TGF-β1/Smad信号通路抑制肝纤维化和肝星状细胞活化[J]. 生理学报 2022; 74 (2): 217-224.

ZHOU Lin-Hua, CHEN Xiao. Geniposide inhibits hepatic fibrosis and hepatic stellate cell activation through blocking the TGF-β1/Smad signaling pathway. Acta Physiol Sin 2022; 74 (2): 217-224 (in Chinese with English abstract).