YTHDF2对肝星状细胞活化增殖与迁移能力的影响  

Effects of YTHDF2 on proliferation and migration of activated hepatic stellate cells

作　　者：孙峰 王娟[1,2] 杨晶晶[2] 周仁鹏[1,2] 胡伟[1,2] 鲁超 SUN Feng;WANG Juan;YANG Jing-jing;ZHOU Ren-peng;Hu Wei;LU Chao(School of Pharmacy,Anhui Medical University,Hefei 230032,China;Dept of Clinical Pharmacology,the Second Hospital of Anhui Medical University,Hefei 230601,China;The First Hospital of Anhui University of Science&Technology,Huainan,Anhui 232001,China)

机构地区：[1]安徽医科大学药学院,安徽合肥230032 [2]安徽医科大学第二附属医院药物临床试验研究中心,安徽合肥230601 [3]安徽理工大学第一附属医院,安徽淮南232001

出　　处：《中国药理学通报》2023年第6期1085-1090,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81600477);安徽省高校自然科学基金资助项目(No KJ2020A0181);安徽医科大学第二附属医院国家自然科学基金孵育计划(No 2019GMFY07);安徽省自然科学基金资助项目(No 2208085MH215)。

摘　　要：目的探究YTHDF2对肝星状细胞(hepatic stellate cells,HSCs)活化增殖与迁移能力的影响。方法使用5μg·L^(-1)TGF-β1诱导HSC-T6细胞活化,使用YTHDF2-siRNA构建YTHDF2沉默模型,实验分为Control组、TGF-β1组、si-NC组和si-YTHDF2组。Dot blot方法检测N^(6)-甲基腺苷(N^(6)-methyladenosine,m^(6)A)水平的变化;Western blot和RT-qPCR方法检测活化HSCs中YTHDF2的表达变化和纤维化关键指标α-SMA、CollagenⅠ的表达变化;CCK-8、Edu染色方法检测HSCs增殖能力;细胞划痕实验和Transwell迁移实验检测HSCs的迁移能力。结果在TGF-β1诱导活化的HSCs中YTHDF2的表达明显增高,而在si-YTHDF2组中水平明显下降;与Control组相比,TGF-β1组中m^(6)A水平以及α-SMA、CollagenⅠ的水平明显升高,且HSCs的增殖和迁移能力也明显增高;而与si-NC组比较,沉默YTHDF2后,m^(6)A水平、α-SMA和CollagenⅠmRNA和蛋白水平均明显降低,同时HSCs的增殖和迁移能力明显被抑制。结论YTHDF2可促进α-SMA和CollagenⅠ的表达,同时可促进HSCs的增殖和迁移能力,进而促进HSCs活化和肝纤维化的进展,这一过程可能是与m^(6)A甲基化修饰有关。Aim To investigate the effect of YTHDF2 on the proliferation and migration of activated hepatic stellate cells(HSCs).Methods 5μg·L^(-1) TGF-β1 was used to induce the activation of HSC-T6 cells,and siRNA was used to construct YTHDF2 silencing model.The experiment was divided into Control group,TGF-β1 group,si-NC group and si-YTHDF2 group.The expressions ofα-SMA and collagenⅠthe levels of YTHDF2 in HSCs were detected by Western blot and RT-qPCR.The levels of m^(6)A was detected by m^(6)A dot blot.Cell proliferation was measured by CCK-8 and Edu staining.The migration ability of HSCs was detected by cell Wound healing assay and Transwell migration assay.Results The level of YTHDF2 increased significantly after TGF-β1 induction while decreased significantly in si-YTHDF2 group.Compared with the Control group,the levels of m^(6)A and the levels ofα-SMA and collagenⅠin TGF-β1 group significantly increased,and the proliferation and migration ability of HSCs were also significantly improved.Compared with the si-NC group,the levels of m^(6)A,the mRNA and protein levels ofα-SMA and collagenⅠwere significantly reduced after YTHDF2 silenced,and the proliferation and migration ability of HSCs were significantly inhibited.Conclusions YTHDF2 promotes the expression ofα-SMA and collagen,and promotes the proliferation and migration ability of HSCs,thereby promoting the activation of HSCs and the progression of liver fibrosis.This process may be related to m^(6)A methylation.

关 键 词：YTHDF2 HSCS m^(6)A 增殖 迁移 肝纤维化 

分 类 号：R-332[医药卫生] R329.28R341R342.2R575.2