Phytomedicine ( IF 6.7 ) Pub Date : 2022-12-07 , DOI: 10.1016/j.phymed.2022.154585 Changlin Du 1 , Jiahui Dong 1 , Qi Wang 1 , Chuanting Xu 1 , Shiqi Feng 1 , Rui Feng 1 , Xiongwen Lv 1 , Jun Li 2 , Lei Zhang 3 , Cheng Huang 1 , Taotao Ma 1
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Background
Hastatoside is an iridoid glycoside extracted from the herb, Verbena officinalis, that exerts various pharmacological effects, including anti-inflammatory, sleep-promoting, and analgesic effects. However, only a few studies have reported the efficacy of hastatoside in liver fibrosis. Liver fibrosis is a pathophysiological process, and its persistence can seriously affect the quality of life and well-being of the patients.
Hypothesis/Purpose
This study aimed to investigate the role of hastatoside on liver fibrosis and its possible underlying mechanisms.
Methods
C57BL/6J mice with carbon tetrachloride (CCl4)-induced hepatic fibrosis were used as the in vivo models. Histological features of the liver were observed using Masson's trichrome and hematoxylin–eosin staining. Alanine aminotransferase and aspartate aminotransferase levels and the hepatic fibrosis indices (type 3 procollagen, laminin, and hyaluronic acid) were measured using corresponding assay kits. LX-2 human hepatic stellate cells (HSCs) stimulated with the transforming growth factor β1 were used as the vitro models. Transfection of the glycogen synthase kinase (GSK)-3β small interfering RNA (siRNA) and β-catenin plasmids was also performed in vitro. Protein levels of GSK-3β, phospho-GSK-3β (Ser 9), α-smooth muscle actin, collagen type I alpha 1, c-Myc, cyclin D1, and β-catenin were determined via western blotting. Moreover, the p-GSK-3β:GSK-3β ratio was calculated to determine the GSK-3β activity.
Results
Hastatoside prevented CCl4-induced liver injury and histological damage. It inhibited the upregulation of α-SMA and Col1α1 levels in a CCl4-induced mouse hepatic fibrosis model. In vitro, hastatoside inhibited the proliferation and activation of HSCs by decreasing the expression levels of cyclin D1 and c-Myc and the proportion of LX-2 cells activated in the G0/G1 phase. Molecular docking results showed that hastatoside bound to GSK-3β. Hastatoside significantly increased the GSK-3β activity and inhibited the downstream effector expression of β-catenin.
Conclusion
These findings suggest that hastatoside can bind to GSK-3β and promote its activity, while inhibiting the GSK-3β downstream effector expression of β-catenin, thereby inhibiting the activation and proliferation of HSCs, which further prevents the development of liver fibrosis. These results provide innovative insights into the underlying liver fibrosis. Moreover, hastatoside is a potential anti-fibrosis monomer that can potentially be used for the treatment of liver fibrosis.
中文翻译：
Hastatoside 通过靶向糖原合酶激酶 3β 减轻四氯化碳诱导的肝纤维化
背景
Hastatoside 是从草本植物Verbena officinalis中提取的一种环烯醚萜苷，具有多种药理作用，包括抗炎、促进睡眠和镇痛作用。然而，只有少数研究报道了海贼苷对肝纤维化的疗效。肝纤维化是一种病理生理过程，其持续存在会严重影响患者的生活质量和幸福感。
假设/目的
本研究旨在探讨藜芦苷对肝纤维化的作用及其可能的潜在机制。
方法
四氯化碳 (CCl 4)-诱导的肝纤维化用作体内模型。使用马松三色和苏木精-伊红染色观察肝脏的组织学特征。使用相应的检测试剂盒测量丙氨酸氨基转移酶和天冬氨酸氨基转移酶水平以及肝纤维化指标（3 型原胶原、层粘连蛋白和透明质酸）。用转化生长因子 β1 刺激的 LX-2 人肝星状细胞 (HSC) 用作体外模型。糖原合酶激酶 (GSK)-3β 小干扰 RNA (siRNA) 和 β-连环蛋白质粒的转染也在体外进行。GSK-3β、磷酸-GSK-3β (Ser 9)、α-平滑肌肌动蛋白、I 型胶原蛋白 α1、c-Myc、细胞周期蛋白 D1 和 β-连环蛋白的蛋白质水平通过蛋白质印迹法测定。此外，p-GSK-3β：
结果
Hastatoside 可防止 CCl 4引起的肝损伤和组织学损伤。它抑制 CCl 4诱导的小鼠肝纤维化模型中 α-SMA 和 Col1α1 水平的上调。在体外，hastatoside 通过降低细胞周期蛋白 D1 和 c-Myc 的表达水平以及在 G0/G1 期激活的 LX-2 细胞的比例来抑制 HSC 的增殖和活化。分子对接结果表明，藜芦苷与 GSK-3β 结合。Hastatoside 显着增加 GSK-3β 活性并抑制 β-catenin 的下游效应子表达。
结论
这些研究结果表明，藜芦苷可以与 GSK-3β 结合并促进其活性，同时抑制 GSK-3β 下游效应子 β-catenin 的表达，从而抑制 HSCs 的活化和增殖，从而进一步阻止肝纤维化的发展。这些结果为潜在的肝纤维化提供了创新的见解。此外，hastatoside 是一种潜在的抗纤维化单体，可用于治疗肝纤维化。
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