骨髓间充质干细胞移植对心肌梗死大鼠心肌钾离子通道蛋白和细胞因子的影响  

Effects of bone marrow mesenchymal stem cell transplantation on voltage-gated K^+ channel proteins and cytokines in the infarcted myocardium of rats

作　　者：胡继宏[1] 赵静苗[1] 王秋萍[1] 贾佳[1] 路娟[1] 金华[1] 侯茜[1] Hu Ji-hong;Zhao Jing-miao;Wang Qiu-ping;Jia Jia;Lu Juan;Jin Hua;Hou Qian(Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China)

机构地区：[1]甘肃中医药大学,甘肃省兰州市730000

出　　处：《中国组织工程研究》2018年第9期1389-1394,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81260597);甘肃省高校基本科研业务费项目(2012-3)~~

摘　　要：背景:研究表明,骨髓间充质干细胞移植可以有效改善心肌梗死后的心功能,移植后的骨髓间充质干细胞可能对心肌电生理有一定影响,但目前相关的研究甚少。目的:观察骨髓间充质干细胞移植对心肌梗死大鼠心肌钾离子电压门控通道蛋白和心肌梗死相关细胞因子的影响,为进一步探索骨髓间充质干细胞治疗心肌梗死引起心律失常的相关机制提供研究基础。方法:将40只SPF级Wistar雄性大鼠随机分为假手术组、模型组、细胞培养液组和骨髓间充质干细胞组。通过冠状动脉左前降支结扎建立心肌梗死模型,并于建模成功15-20 min后在梗死区周围采用四点法注射100μL骨髓间充质干细胞悬液(含1×10~6个骨髓间充质干细胞)和100μL细胞培养液。细胞移植4周后获取心脏标本,采用苏木精-伊红染色观察梗死心肌组织病理形态并计算梗死面积,采用Westernn blot检测钾离子电压门控通道蛋白Kv1.2和Kv1.5和心肌细胞特异性蛋白cTnT的表达,利用免疫组化方法检测梗死区心肌细胞凋亡因子Caspase-3、自噬因子Bcl-2和一氧化氮及超氧化物歧化酶的表达。结果与结论:(1)与模型组相比,骨髓间充质干细胞组心肌梗死面积减小(P<0.05),cTnT、Kv1.5和超氧化物歧化酶表达水平均提高(P<0.05),Caspase-3、Bcl-2和Kv1.2表达水平均降低(P<0.05);(2)与细胞培养液组比较,骨髓间充质干细胞组心肌梗死面积减小(P<0.05),cTnT、Kv1.5和超氧化物歧化酶表达水平均提高(P<0.05),Caspase-3、Bcl-2和Kv1.2表达水平均降低(P<0.05);(3)结果提示,骨髓间充质干细胞移植提高了心肌抗氧化能力,减少了细胞凋亡和自噬,促进钾离子通道蛋白Kv1.5的表达。Abstract BACKGROUND:Studies have shown that bone marrow mesenchymal stem cell(BMSC)transplantation can effectively improve cardiac function after myocardial infarction.However,few reports have been issued on myocardial electrophysiology after BMSCs transplantation.OBJECTIVE:To observe the effects of BMSCs transplantation on voltage-gated K+channel protein and myocardial infarction-related cytokines,thereby providing basic evidence for further exploration on the mechanism underlying arrhythmia in myocardial infarction due to BMSCs transplantation.METHODS:Forty male Wistar rats,SPF grade,were randomly divided into four groups:sham group,model group,cell culture medium group and BMSCs group.The myocardial infarction model was created in rats by permanent ligation of the left descending coronary artery.At 15-20 minutes after surgery,BMSCs(100μL,1×106)or cell culture medium(100μL)was injected at four sites in the peri-infarct zone.Four weeks after cell therapy,cardiac samples were taken,the pathological morphology of the infarcted myocardium was observed by hematoxylin-eosin staining,and the infarct size was calculated;the expression levels of voltage-gated K+channel proteins Kv1.2 and Kv1.5 and cardiac troponin T(cTnT)were measured by western blot assay;and the expression levels of apoptotic factor(Caspase-3),autophagy factor(Bcl-2),nitric oxide and superoxide dismutase were tested by immunohistochemistry.RESULTS AND CONCLUSION:Compared with the model group and cell culture medium group,the infarct size decreased in the BMSCs group(P<0.05);the expression levels of cTnT,Kv1.5 and superoxide dismutase increased(P<0.05),and the expression levels of Caspase-3,Bcl-2 and Kv1.2 decreased(P<0.05)in the BMSCs group.In summary,BMSCs transplantation can promote the expression of voltage-gated K+channel proteins,and improve anti-oxidation capacity of the myocardium and decrease apoptosis and autophagy.

关 键 词：骨髓 间质干细胞移植 心肌梗塞 钾通道 电压门控 细胞凋亡 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]