丹酚酸B抑制HuH-7细胞的Hippo/YAP通路机制研究  

Study on mechanism of Hippo/YAP pathway inhibitionby salvianolic acid B in HuH-7 cells

作　　者：余新梅 李利利 张傲 邓洁 杨雁[1] YU Xin-mei;LI Li-li;ZHANG Ao;DENG Jie;YANG Yan(School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China;First Clinical Medical College,Anhui Medical University,Hefei 230032,China;Second Clinical Medical College,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学基础医学院,安徽合肥230032 [2]安徽医科大学第一临床医学院,安徽合肥230032 [3]安徽医科大学第二临床医学院,安徽合肥230032

出　　处：《中国药理学通报》2024年第1期106-113,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 82074073);安徽医科大学临床医学专业“早期接触科研”训练计划立项项目(No 2021-ZQKY-64,No 2022-ZQKY-175)。

摘　　要：目的 探讨丹酚酸B(salvianolic acid B,Sal B)对人肝癌HuH-7细胞是否具有抑制作用,并探讨其是否通过Hippo/YAP信号通路发挥作用。方法 采用TGF-β1 (9 pmol·L^(-1))或MST1/2抑制剂XMU-MP-1 (5、10μmol·L^(-1))刺激HuH-7细胞,用Sal B(50μmol·L^(-1))处理HuH-7细胞。用CCK-8和EdU检测细胞增殖情况,用细胞划痕实验检测细胞迁移情况,用免疫荧光染色法检测YAP和TAZ的表达和分布,用Western blot检测p-MST1、MST1、p-YAP、YAP、p-TAZ、TAZ和CTGF的蛋白表达水平。结果 Sal B抑制了TGF-β1或XMU-MP-1刺激的HuH-7细胞的增殖。同时,Sal B抑制了TGF-β1刺激的HuH-7细胞的迁移。值得注意的是,与XMU-MP-1对HepG2细胞迁移能力的促进作用不同,XMU-MP-1在本实验中不能明显促进HuH-7细胞的迁移。此外,Sal B可上调p-MST1、MST1、p-YAP、p-TAZ的蛋白表达,降低YAP、TAZ、CTGF的表达。结论 Sal B抑制HuH-7细胞的作用可能与激活Hippo/YAP信号通路有关。Aim To investigate whether salvianolic acid B(Sal B)has inhibitory effect on hepatoma HuH-7 cells and explore whether it works via Hippo/YAP signaling pathway.Methods HuH-7 cells were induced by TGF-β1(9 pmol·L^(-1))or the MST1/2 inhibitor,XMU-MP-1(5,10μmol·L^(-1))and pretreated with Sal B(50μmol·L^(-1)).CCK8 and EdU assays were used to evaluate cell proliferation.Wound healing assay was used to detect cell migration.Immunofluorescence was used to evaluate the expression and distribution of YAP and TAZ.Western blot was used to detect the levels of protein expression of p-MST1,MST1,p-YAP,YAP,p-TAZ,TAZ,and CTGF.Results The results showed that Sal B inhibited the proliferation of TGF-β1 or XMU-MP-1-stimulated HuH-7 cells.At the same time,Sal B suppressed the migration of TGF-β1-stimulated HuH-7 cells.Notably,unlike the promotion of XMU-MP-1 on the migration ability of HepG2 cells,XMU-MP-1 could not significantly promote the migration of HuH-7 cells in this experiment.In addition,Sal B increased protein expressions of p-MST1,MST1,p-YAP and p-TAZ,while decreased the expressions of YAP,TAZ,CTGF.Conclusions The inhibitory effect of Sal B on HuH-7 cells may be related to the activation of Hippo/YAP signaling pathway.

关 键 词：丹酚酸B HuH-7 肝癌 Hippo/YAP TGF-Β1 XMU-MP-1 

分 类 号：R967[医药卫生—药理学] R392.5[医药卫生—药学] R392.12