高迁移率族蛋白1及其受体对大鼠骨髓间充质干细胞增殖及细胞因子水平的影响  

Effects of high mobility group box 1 and its receptor on proliferation and cytokine levels of rat marrow mesenchymal stem cells

作　　者：刘志江[1] 石蓓[1] 束波[2] 马帅[1] LIU Zhi-Jiang SHI Bei SHU Bo MA Shuai(Department of Cardiology, the First Affiliated Hospital of Zunyi Medical University Department of Biochemistry and Molecular Biology of Zunyi Medical University, Zunyi, Guizhou 563000, China)

机构地区：[1]遵义医学院第一附属医院心内科,贵州省遵义市563000 [2]遵义医学院生物化学与分子生物学教研室,贵州省遵义市563000

出　　处：《中国动脉硬化杂志》2017年第4期337-342,共6页Chinese Journal of Arteriosclerosis

基　　金：贵州省科技计划项目[黔科合SY字(2012)3109号];遵义市科技计划项目[遵市科会社字(2014)82号]

摘　　要：目的探讨高迁移率族蛋白1(HMGB1)及其受体对大鼠骨髓间充质干细胞(MSC)增殖及细胞因子水平的影响。方法体外传代培养大鼠MSC,以25.0、50.0及100.0μg/L HMGB1分别作用细胞24 h、48 h、72 h后,MTT检测MSC的增殖情况。25.0、50.0及100.0μg/L HMGB1作用MSC 48 h后,ELISA测定细胞培养液中血管内皮生长因子(VEGF)和碱性成纤维细胞生长因子(b FGF)水平,Western blot检测MSC中HMGB1的晚期糖基化终产物受体(RAGE)和Toll样受体4(TLR4)的表达情况。确定HMGB1对MSC作用的最佳药物浓度后,siRNA分别抑制RAGE受体和TLR4受体,观察HMGB1对MSC增殖及分泌细胞因子水平的影响。结果 25.0μg/L HMGB1作用MSC 24 h、48 h、72 h后均能有效促细胞增殖(P<0.05);25.0、50.0μg/L HMGB1作用细胞48 h后,细胞释放VEGF、b FGF明显增多,RAGE及TLR4表达也增高(P<0.05)。特异siRNA转染靶向干扰MSC中TLR4的表达后,促细胞增殖及分泌VEGF、b FGF作用受到抑制(P<0.05),而siRNA干扰RAGE表达对HMGB1的细胞效应无明显影响。结论 HMGB1可通过结合其受体TLR4促进MSC增殖以及VEGF、b FGF分泌。Aim To investigate the effect of high mobility group box 1 （HMGB1） and its receptor on survival and cytokine levels of rat marrow mesenchymal stem cells. Methods Marrow mesenchymal stem cells were treated with HMGB1 at 25.0, 50.0 and 100.0 μg/L for 24, 48 and 72 hours. Cell proliferations were detected by MIT after different treatment. VEGF and bFGF were detected by ELISA. Expressions of receptor of advanced glycation end products （RAGE） and Toll-like receptor 4 （TLR4） in mesenchymal stem cells were detected by Western blot. Real-time PCR and Western blot were used to examine RAGE and TLR4 expression, which were to verify the efficiency of transfection with siRNA in marrow mesenchymal stem cells. MTT assay and ELISA were used to analyze proliferation and secretions of VEGF and bFGF in si-RAGE or si-TLR4 marrow mesenchymal stem cells. Results 25.0 μg/L HMGB1 could increase marrow mesenchymal stem cells proliferation （P〈0.05）. 25.0 and 50.0 μg,/L HMGB1 could promote VEGF and bFGF secretion compared with control group （P〈0.05）. The protein levels of RAGE and TLR4 in 25.0 μg/L HMGBI group were significantly higher than those in other groups （P〈0.05）. Marrow mesenchymal stem cells transfected with siTLR4 antagonized the cell proliferation and secretion of VEGF, bFGF induced by HMGB1. Conclusion HMGB1 promote cell proliferation and secret VEGF, bFGF by binding to its TLR4 receptor.

关 键 词：高迁移率族蛋白1 骨髓间充质干细胞 细胞因子 

分 类 号：R363[医药卫生—病理学]