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

发布时间：2018-01-27 06:09

本文关键词： 高迁移率族蛋白骨髓间充质干细胞细胞因子　出处：《中国动脉硬化杂志》2017年04期 　论文类型：期刊论文

【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(P0.05);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(P0.05)。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(P0.05),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。
[Abstract]:Objective to investigate the effect of high mobility group protein 1 (HMGB1) and its receptor on rat bone marrow mesenchymal stem cells (MSC) proliferation and cytokine levels. Methods in vitro cultured rat MSC, 25.0,50.0 and 100 g/L HMGB1 respectively. Cell 24 h, 48 h, 72 h, MTT detection the proliferation of MSC.25.0,50.0 and 100 g/L HMGB1 MSC 48 h, ELISA was determined in cell culture fluid vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (B FGF), HMGB1 Western blot advanced glycation endproducts in detection of MSC receptor (RAGE) and Toll like receptor 4 (TLR4). The expression of drug effect on MSC to determine the optimal concentration of HMGB1, siRNA respectively inhibited RAGE and TLR4 receptors, the effect of HMGB1 on proliferation and cytokines secretion of MSC. The 25 g/L HMGB1 MSC 24 h, 48 h, 72 h were found to be effective in promoting cell proliferation (P0.0 5); 25.0,50.0 g/L HMGB1 cells after 48 h cells to release VEGF, B FGF increased significantly, RAGE and TLR4 expression also increased (P0.05). Specific siRNA targeting expression of TLR4 interference in MSC, promoting cell proliferation and secretion of VEGF, B FGF (P0.05) was inhibited, whereas no obvious effect of siRNA interference RAGE expression effects on HMGB1. Conclusion HMGB1 can promote the proliferation of MSC binding to its receptor TLR4, VEGF, B and FGF secretion.

【作者单位】：遵义医学院第一附属医院心内科;遵义医学院生物化学与分子生物学教研室;
【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]
【分类号】：R542.22
【正文快照】： 骨髓间充质干细胞(marrow mesenchymal stemcells,MSC)具有多向分化潜能,MSC移植治疗心肌梗死的有效性目前已得到基础和临床研究的证实[1],但分子机制尚不清楚。研究发现MSC的旁分泌效应在梗死心肌的修复中发挥了重要作用。高迁移率族蛋白B1(high mobility group box 1,HMGB1)

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