摘要
目的 探讨普罗布考在保护人脂肪间充质干细胞（hADSCs）对抗高糖引起损伤中的作用及机制.方法 原代培养人脂肪间充质干细胞hADSCs,应用免疫荧光对3～5代细胞表面抗原CD34、CD45、CD90、CD105进行表型鉴定.将hADSCs分为正常糖对照组（CON组）、高糖组（GLU组）、普罗布考＋高糖组（PRO＋GLU组）、抑制剂SB203580＋高糖组（SB＋GLU组）、普罗布考组（PRO组）、抑制剂SB203580组（SB组）.MTT法测定细胞增殖;流式细胞仪检测细胞凋亡情况;DCFH-DA法检测细胞内活性氧水平.Western blot技术检测p-p38MAPK蛋白的表达水平.结果 与CON组比较,GLU组hADSCs细胞增殖活性明显降低（P＜0.05）,凋亡细胞比例、ROS水平显著增加（P＜0.05）;与GLU组比较,PRO＋GLU组和SB＋GLU组hADSCs细胞细胞增殖活性都明显增加（P＜0.05）,凋亡细胞比例、ROS水平表达水平均显著降低（P＜0.05）;与CON组比较,GLU组p-p38MAPK表达水平明显上升,普罗布考预处理能明显抑制高糖诱导的p-p38MAPK表达水平上调.结论 普罗布考能帮助hADSCs对抗高糖引起的损伤;p38MAPK通路参与高糖对hADSCs的损伤作用;普罗布考可通过抑制p38MAPK通路的激活保护hADSCs对抗高糖诱导的损伤.
Objective To investigate the effect of probucol on high glucose-induced impairment in human adipose mesenchymal stem cells （hAD-SCs）.Methods hADSCs were isolated from human adipose tissue.The expression profiles of CD34,CD45,CD90,and CD 105 were examined by immunofluorescence.The hADSCs were treated with normal glucose,high glucose,probucol with high glucose,SB203580 with high glucosse,probucol and SB203580,respectively.The proliferation of hADSCs was determined by MTT assay.Apoptotic cells were analyzed using flow cytometry.Intracellular ROS levels were measured by using DCFH-DA.The expression level of p-p38MAPK protein was tested by Western blot.Results Compared with normal control group,high glucose decreased hADSC proliferative potential,increased the amount of apoptotic cells and intracellular ROS levels,upregulated expression level of p-p38MAPK protein.Compared with high glucose group,pretreatment with probucol increased hADSC proliferative potential,decreased in amount of apoptotic cells and intracellular ROS levels,obviously inhibited high glycose induced upregulation of pp38MAPK expression.Similar to the protective effect of probucol,pretreatment with SB203580 （an inhibitor of p38MAPK） also protected hADSCs against high glucose-induced injury.Conclusion Probucol can protect hADSCs against high glucose-induced injury; p38MAPK pathway was involved in glucose-induced ADSC injury; Probucol may protect ADSC against high glucose-induced impairment by medulating p38MAPK pathway.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2014年第7期615-620,共6页
Journal of China Medical University
基金
辽宁省科学技术计划(2011225020
2012225014
2013225303)
沈阳市科技局科技计划(F11-262-9-52)
