YTHDF1调控Fis1对肝星状细胞活化、增殖及迁移能力的影响
YTHDF1 regulation of Fis1 on the activation and proliferation and migration ability of hepatic stellate cells
摘要
Abstract
Objective To explore the effect of YTH domain family protein 1(YTHDF1)on the activation,prolifer-ation and migration of hepatic stellate cells(HSCs)by regulating mitochondrial fission mediated by mitochondrial fission protein 1(Fis1).Methods The mouse hepatic stellate cell line JS-1 was treated with 5 ng/ml TGF-β1 for 24 h to induce its activation and proliferation,and YTHDF1-siRNA was used to construct a YTHDF1 silencing mod-el.The experiment was divided into Control group,TGF-β1 group,TGF-β1+si-NC group and TGF-β1+si-YTH-DF1 group.Expression changes of YTHDF1,Fis1 and key indicators of fibrosis,type Ⅰ collagen(Collagen Ⅰ)and α-smooth muscle actin(α-SMA)were detected through reverse transcription quantitative polymerase chain re-action(RT-qPCR)and Western blot;CCK-8 was used to detect cell proliferation ability;Trans well migration assay and cell scratch assay were used to detect cell migration ability;immunofluorescence staining experiment was used to detect the effect of YTHDF1 on Fis1-mediated mitochondrial fission;finally,JC-1 staining was used to experi-mentally detect the effect of YTHDF1 on mitochondrial membrane potential.Results Compared with the Control group,RT-qPCR and Western blot experimental results showed that the expression of YTHDF1 and Fis1 increased in the TGF-β1 group(P＜0.05,P＜0.01;P＜0.000 1),as well as the fibrosis markers Collagen Ⅰ and the ex-pression level of α-SMA increased(P＜0.01;P＜0.001,P＜0.000 1);while adding CCK-8,the experimental results showed that the proliferation ability of HSCs in the TGF-β1 group was enhanced(P＜0.000 1);Trans well experimental results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P＜0.01);the cell scratch experiment results showed that the migration ability of HSCs in the TGF-β1 group was enhanced(P＜0.000 1);the immunofluorescence experiment results showed that the TGF-β1 group Mito-Tracker Red staining and Fis1 co-localization signal increased(P＜0.05);JC-1 staining experiment results showed that the mitochondri-al membrane potential increased in the TGF-β1 group(P＜0.01).Compared with the TGF-β1+si-NC group,RT-qPCR and Western blot experimental results showed that the expression of YTHDF1 and Fis1 in the TGF-β1+si-YTHDF1 group was reduced(P＜0.01;P＜0.001),and fibrosis markers the levels of Collagen Ⅰ and α-SMA were reduced(P＜0.01;P＜0.001,P＜0.01).CCK-8 experimental results showed that the proliferation ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P＜0.000 1);Transwell experiment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P＜0.001);cell scratch ex-periment results showed that the migration ability of HSCs in the TGF-β1+si-YTHDF1 group was weakened(P＜0.000 1);immunofluorescence experiment results showed that the Mito-Tracker Red staining and Fis1 co-localiza-tion signal decreased in the TGF-β1+si-YTHDF1 group(P＜0.01);JC-1 staining experiment results showed that mitochondrial membrane potential decreased in the TGF-β1+si-YTHDF1 group(P＜0.05).Conclusion YTH-DF1 promotes the activation,proliferation and migration capabilities of HSCs by positively regulating Fis1-mediated mitochondrial fission.This suggests that YTHDF1 may be a key gene involved in regulating the activation,prolifera-tion and migration of HSCs.关键词
YTHDF1/Fis1/HSCs/活化/增殖/迁移/肝纤维化Key words
YTHDF1/Fis1/HSCs/activation/proliferation/migration/hepatic fibrosis分类
医药卫生引用本文复制引用
贾琳,孙峰,董琪琪,杨晶晶,周仁鹏,胡伟,鲁超..YTHDF1调控Fis1对肝星状细胞活化、增殖及迁移能力的影响[J].安徽医科大学学报,2025,60(1):49-58,10.基金项目
国家自然科学基金资助项目(编号:81600477) （编号:81600477）
安徽高校自然科学基金资助项目(编号:KJ2020A0181) （编号: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(编号:2019GMFY07) （编号:2019GMFY07）
安徽省重点研究与开发计划项目(编号:202204295107020035) （编号:202204295107020035）
安徽省自然科学基金面上项目(编号:2208085MH215) （编号:2208085MH215）
合肥综合性国家科学中心大健康研究院职业医学与健康联合研究中心项目(编号:OMH-2023-03) National Natural Science Foundation of China(No.81600477) （编号:OMH-2023-03）
Natural Science Foundation for Universities in Anhui Province(No.KJ2020A0181) （No.KJ2020A0181）
National Natural Science Foundation Incubation Plan of the Second Affiliated Hospital of Anhui Medical University(No.2019GMFY07) （No.2019GMFY07）
Key Research and Development Pro-gram of Anhui Province(No.202204295107020035) （No.202204295107020035）
Natural Science Foundation of Anhui Province(No.2208085 MH215) （No.2208085 MH215）
Occupational Medicine and Health Joint Research Project from Institute of Health and Medicine,Hefei Comprehensive National Science Center(No.OMH-2023-03) （No.OMH-2023-03）
