小鼠骨髓间充质干细胞通过JAK2/STAT3信号通路对成纤维细胞增殖和胶原表达水平的影响  

Effect of bone marrow mesenchymal stem cells of mice on proliferation and collagen expression levels of fibroblasts through JAK2/STAT3 signaling pathway

作　　者：黎涵玥 阳莲 刘剑锋[1] 张舒飞 洪莉[1] LI Hanyue;YANG Lian;LIU Jianfeng;ZHANG Shufei;HONG Li(Department of Obstetrics and Gynecology,Renmin Hospital,Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院妇产科,湖北武汉430060

出　　处：《吉林大学学报(医学版)》2025年第2期325-332,共8页Journal of Jilin University:Medicine Edition

基　　金：国家自然科学基金项目(82371639);国家重点研发计划项目(2021YFC2701300,2021YFC2701302);湖北省科技厅自然科学基金项目(2022CFB124);湖北省科技厅重点研发项目(2022BCA045);湖北省领军人才项目([2019] 47)。

摘　　要：目的:探讨骨髓间充质干细胞(BMSCs)对L929细胞增殖和其胶原表达水平的影响,阐明其相关作用机制。方法:提取4周龄C57BL/6小鼠的BMSCs。采用免疫荧光染色鉴定BMSCs的表型。将L929细胞分为对照组(L929细胞)、共培养组(L929细胞与BMSCs)、Janus激酶(JAK)抑制剂WP1066组(WP1066处理L929细胞与BMSCs)和二甲基亚砜(DMSO)组(DMSO处理L929细胞与BMSCs)。细胞计数试剂盒8 (CCK-8)法检测不同时间点各组L929细胞增殖活性,Western blotting法检测各组L929细胞中Ⅰ型胶原蛋白(ColⅠ)和Ⅲ型胶原蛋白(ColⅢ)表达水平,免疫荧光染色法检测各组L929细胞中ColⅠ和ColⅢ蛋白表达。结果:BMSCs表面抗原(SA)荧光检测,BMSCs表达表面标志物CD29+、CD45-、CD90+和CD105+。CCK-8法检测,与对照组比较,共培养组L929细胞增殖活性明显升高(P<0.01),DMSO组L929细胞增殖活性明显升高(P<0.01);与共培养组比较,WP1066组L929细胞增殖活性明显降低(P<0.01)。Western blotting法检测,与对照组比较,共培养组和DMSO组L929细胞中ColⅠ及ColⅢ蛋白表达水平明显升高(P<0.01);与共培养组比较,WP1066组L929细胞中ColⅠ和ColⅢ蛋白表达水平明显降低(P<0.01);与DMSO组比较,WP1066组L929细胞中ColⅠ和ColⅢ蛋白表达水平明显降低(P<0.01)。免疫荧光染色法检测,与对照组比较,共培养组和DMSO组L929细胞中ColⅠ及ColⅢ荧光强度明显升高(P<0.01);与共培养组比较,WP1066组L929细胞中ColⅠ和ColⅢ荧光强度明显降低(P<0.01);与DMSO组比较,WP1066组L929细胞中ColⅠ和ColⅢ荧光强度明显降低(P<0.01)。结论:间充质干细胞可通过JAK2/信号转导和转录激活子3 (STAT3)信号通路促进小鼠L929细胞增殖及胶原生成。Objective:To investigate the effect of bone marrow mesenchymal stem cells(BMSCs)on the proliferation and collagen expression levels of L929 cells,and to clarify its related mechanism.Methods:The BMSCs were extracted from the 4-week-old C57BL/6 mice.The phenotypes of BMSCs were identified by immunofluorescence staining.The L929 cells were divided into control group(L929 cells),co-culture group(L929 cells and BMSCs),inhibitor of Janus kinase(JAK)WP1066 group(WP1066-treated L929 cells and BMSCs),and dimethyl sulfoxide(DMSO)group(DMSO-treated L929 cells and BMSCs).Cell counting kit-8(CCK-8)assay was used to detect the proliferation activities of the L929 cells in various groups at different time points;Western blotting method was used to detect the expression levels of typeⅠcollagen(ColⅠ)and typeⅢcollagen(ColⅢ)in the L929 cells in various groups;immunofluorescence staining was used to detect the expressions of ColⅠand ColⅢproteins in the L929 cells in various groups.Results:The fluorescence assay results of surface antigen(SA)showed that the surface markers CD29+,CD45-,CD90+and CD105+were found in the BMSCs.The CCK-8 assay results showed that compared with control group,the proliferation activities of the L929 cells in co-culture group and DMSO group were significantly increased(P<0.01);compared with co-culture group,the proliferation activity of the L929 cells in WP1066 group was significantly decreased(P<0.01).The Western blotting method results showed that compared with control group,the expression levels of ColⅠand ColⅢproteins in the L929 cells in co-culture group and DMSO group were significantly increased(P<0.01);compared with co-culture group,the expression levels of ColⅠand ColⅢproteins in the L929 cells in WP1066 group were significantly decreased(P<0.01);compared with DMSO group,the expression levels of ColⅠand ColⅢproteins in the L929 cells in WP1066 group were significantly decreased(P<0.01).The immunofluorescence staining results showed that compared with control group,the fluore

关 键 词：盆底功能障碍性疾病 骨髓间充质干细胞 成纤维细胞 Ⅰ型胶原蛋白 Ⅲ型胶原蛋白 JANUS激酶2 信号转导和转录激活子3 

分 类 号：R711[医药卫生—妇产科学]