摘要
目的探讨人脐带具有心肌特异分化潜能的间充质干细胞外泌体(MSCs-exosomes)对大鼠心肌梗死冠状动脉再通后损伤过程中诱导凋亡的干预作用以及对磷脂酰肌醇-3激酶/蛋白激酶B (PI-3K/AKT)通路的影响。方法将40只SD大鼠随机分为假手术组(S组)、缺血再灌注组(I/R组)、外泌体干预组(Ex组)、外泌体干预联合PI-3K抑制剂组(Ex+L组)。再灌注后,采用酶联免疫吸附实验(ELISA)测定血清乳酸脱氢酶(LDH),细胞计数试剂盒(CCK-8)分析评估细胞活力,原位缺口末端标记(TUNEL)法检测心肌凋亡率,蛋白免疫印迹法(Western-blot)检测凋亡相关蛋白Bcl-2、Bax、Caspase-3以及Caspase-9和细胞色素C(Cyt C)的表达水平。结果与S组相比,I/R组心肌细胞Bax、Caspase-3、Caspase-9、Cyt C表达水平增高,凋亡比例明显增高,血清中LDH水平明显上升。Ex组可明显减少细胞损伤,降低促凋亡相关蛋白的水平,促使血清中LDH含量下降。结论干细胞外泌体可减轻心肌梗死冠状动脉再通后损伤,这种保护机制可能与PI-3K/AKT调控的线粒体凋亡途径有关。
Objective To explore the intervention effect of human umbilical cord mesenchymal stem cells exosomes(MSCs-exosomes) on the induction of apoptosis and the effect on PI-3K/AKT signaling pathway in the process of reperfusion after myocardial infarction in rats.Methods Forty SD rats were randomly divided into sham operation group(group S),ischemia-reperfusion group(group I/R),exosomes intervention group(Ex group),exosomes intervention combined with PI-3K inhibitor group(Ex+L group).After reperfusion,serum lactate dehydrogenase(LDH) was measured by ELISA assay,cell viability was analyzed by CCK-8 kit,TUNEL method was used to detect the apoptosis rate of myocardium,Western blot was used to detect the expression level of apoptosis related protein(Bcl-2,Bax),Caspase-3,Caspase-9 and cyt C.Results Compared with group S,The expression level of Bax、Caspase-3、Caspase-9、Cyt C increased significantly in group I/R,and the proportion of apoptosis increased significantly and the level of LDH in serum increased significantly.The Ex group could significantly reduce the cell damage,reduce the level of apoptosis related protein and decrease the content of LDH in the serum.Conclusion the stem cell exosomes can reduce the injury of reperfusion after myocardial infarction,which may be related to the mitochondrial apoptosis pathway regulated by PI-3K/AKT.
引文
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