摘要
目的探讨严重烧伤大鼠血清对骨髓间充质干细胞(BMSCs)迁移的影响及作用机制。方法建立严重烧伤大鼠模型,并制备严重烧伤大鼠血清。实验分为正常培养组(含10%胎牛血清,C组)、烧伤血清组(含10%烧伤大鼠血清,B组)、烧伤血清+阻断剂组(含10%烧伤大鼠血清+终浓度为10μmol/L的PI3K信号通路抑制剂LY294002,B+LY组),MTT法检测各组细胞活性;Transwell小室检测各组细胞迁移情况;Western blot检测各组细胞p-Akt、Akt蛋白表达;免疫荧光检测各组细胞微管结构。结果与C组比较,B组烧伤血清处理24h后,BMSCs细胞活性增高(P<0.01),p-Akt水平升高(P<0.05),细胞微管结构出现解聚,迁移数量增高(P<0.001);加入抑制剂后,与B组比较,B+LY组BMSCs细胞活性降低(P<0.01),迁移数量降低(P<0.001),p-Akt水平降低(P<0.05),细胞微管结构变化不明显。结论严重烧伤大鼠血清可促进BMSCs细胞迁移,可能与烧伤血清中细胞因子激活PI3K/Akt信号通路相关,进而引起BMSCs细胞微管结构变化,促进BMSCs迁移。
Objective To study the effects of severely burned rats serum on migration of BMSCs and mechanism.MethodsSeverely burned rats model was established,and the preparation of severely burned rats serum.Experimental groups：normal training group（containing 10%fetal bovine serum,group C）,burn serum group（containing 10% burns in the rat serum,group B）,burn serum＋blockers（10% burns in the rat serum＋final concentration of 10μmol/L PI3 Ksignaling pathway inhibitor LY294002 training,group B＋LY）.Activity of cells was examined with MTT;migration of cells was examined with Transwell chambers testing;protein expression of p-AKT/AKT was determined with Western blot;microtubule structure of cells was examined with immunofluorescence.Results Compared with group C,group B burn serum treatment after 24 h,BMSCs activity（P〈0.01）,p-AKT levels（P〈0.05）,increased migration quantity（P〈0.001）;cell microtubule structures appear rupture,after adding inhibitor,compared with group B,group B＋LY BMSCs activity（P〈0.01）,to reduce the number of migration（P〈0.001）,p-lower AKT（P〈0.05）,cell microtubule structure similar to the normal group.Conclusion Severely burned rats serum can promote BMSCs migration,may burn serum cytokine activation of PI3K/AKT signal pathway,resulting in cell microtubule structure change,promote the migration of BMSCs.
出处
《重庆医学》
CAS
北大核心
2015年第27期3752-3755,共4页
Chongqing medicine
基金
重庆市基础与前沿研究计划项目(cstc2013jcyjA10069)
