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干扰Nrf2基因后骨髓间充质干细胞移植对大鼠心肌梗死后心肌凋亡的影响

, PP. 1319-1324

Keywords: NF-E相关因子,siRNA,心肌梗死,骨髓间充质干细胞,凋亡

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Abstract:

目的观察干扰核因子相关因子2(Nrf2)的骨髓间充质干细胞(mesenchymalstemcells,MSCs)注射大鼠心肌梗死局部后细胞凋亡情况及对心功能的影响。方法建立心肌梗死大鼠模型,随机分为:①干扰Nrf2基因的MSCs移植(Nrf2干扰组)组;②空载慢病毒修饰的MSCs移植(Nrf2正常组);③生理盐水注射对照组(对照组,n=12)。心肌梗死细胞移植后7d,细胞免疫荧光法检测心肌局部caspase3蛋白的表达;移植后28dHE染色评价梗死后心肌组织病理变化,TTC染色评估梗死面积;采用蛋白印迹法(Westernblot)检测心肌梗死区Nrf2和凋亡蛋白Bcl2、Bax的蛋白表达水平。结果心梗细胞移植后7d,心肌梗死区域有EGFP标记的MSCs,部分EGFP表达位置检测到Caspase3表达阳性,Nrf2干扰组凋亡蛋白Caspase3的荧光强度强,Nrf2正常组Caspase3的荧光强度弱,对照组无EGFP表达;细胞移植后28d,HE染色结果显示,与Nrf2正常组比较,Nrf2干扰组心肌组织肌纤维排列紊乱,炎性细胞浸润明显,而Nrf2正常组细胞排列整齐,炎性细胞浸润减少;心肌TTC染色结果显示,Nrf2干扰组的心肌梗死面积为(35.30±0.41)%,较Nrf2正常组[(25.07±0.15)%]明显增加(P<0.05),与对照组[(35.40±0.70)%]比较无差异(P>0.05);Westernblot结果显示,与Nrf2正常组比较,Nrf2干扰组心肌中Nrf2蛋白和抗凋亡蛋白Bcl-2表达下降,而促凋亡蛋白Bax表达增加,差异有统计学意义(P<0.05),Nrf2干扰组与对照组比较,各蛋白表达无差异性(P>0.05);心功能改善方面,Nrf2干扰组EF[(40.83±2.67)%]较Nrf2正常组[(52.35±1.58)%]降低(P<0.050而心室容积较[(LVDd,(1.090±0.061),LVDs(0.750±0.062)]较Nrf2正常组[(LVDd,(0.7900±0.0245),LVDs(0.610±0.026)]增加。结论移植干扰Nrf2的MSCs后,心肌梗死区凋亡相关蛋白表达增加,降低了外源性MSCs对梗死心脏的修复能力。

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