微泡破坏及间充质干细胞治疗大鼠心肌损伤模型的有效性研究

Study on the validation of microbubble destruction and MSC in treating myocardial damage model of rat

目的:探究超声微泡介导间充质干细胞(MSC)移植治疗阿霉素心肌病的效果。方法:取SD大鼠36只,建立阿霉素心肌病模型,采用随机数表法将其分为磷酸盐缓冲液(PBS)组、单纯MSC组和超声微泡+MSC组,每组12只。采用1.0 W/cm^2、2.0 W/cm^2和3.0 W/cm^2不同强度超声微泡对大鼠MSC干预与检测。采用SPIO标记、荧光标记、蛋白印迹法等分别对3组大鼠进行心肌组织学检查、心功能检查、心肌bax、bcl-2蛋白表达及移植后24 h的MSC心肌归巢阳性比率进行比较。结果:①1.0 W/cm^2、2.0 W/cm^2和3.0 W/cm^2强度超声辐射90 s/d、120 s/d辐射后,即刻、24 h和48 h的MSC吸光度水平比较,差异均有统计学意义(F=9.314,F=8.584,F=9.026;P<0.05),且1.0 W/cm^2即刻、24 h和48 h的MSC吸光度水平显著高于2.0 W/cm^2和3.0 W/cm强度超声辐射;②三组大鼠的bax蛋白、bcl-2蛋白和bcl-2/bax表达水平比较,差异均有统计学意义(F=9.333,F=7.962,F=29.514;P<0.05);三组大鼠移植部位毛细血管数、荧光标记细胞数比较差异均有统计学意义(F=25.043,F=36.697;P<0.05);③移植后24h超声微泡+MSC组检出MSC心肌归巢阳性率最高,三组检出MSC心肌归巢阳性率比较差异有统计学意义(F=166.411,P<0.05)。结论:超声辐射微泡可促进阿霉素心肌病大鼠自体骨髓间充质干细胞(BM-MSC)的移植,促进心肌血管新生,改善因阿霉素心肌梗死而受损的心功能。

Objective:To explore the effect of transplantation therapy ultrasound microbubble-mediated mesenchymal stem cell(MSC)in treating Adriamycin cardiomyopathy.Methods:Thirty-six SD rats were selected to establish Adriamycin cardiomyopathy model,and they were divided into phosphate buffered saline(PBS)group,simple MSC group and ultrasound microbubble+MSC group by adopting random number table method,with 12 rats in each group.And the ultrasound microbubbles with different intensity included 1.0 w/cm2,2.0 W/cm2 and 3.0 W/cm2 were adopted to intervene and detect MSC of rats.And superparamagnetic iron oxide(SPIO)labeling,fluorescent labeling,Western blotting,etc.were adopted respectively to implement histological examination of cardiac muscle,cardiac functional examination,the expression of bax and bc1-2 protein of myocardium and the calculation of positive ratio of MSC myocardial homing at 24 h post transplantation.Results:1)The differences of the MSC absorbance levels at once,24 h and 48 h post ultrasound radiation of 90 s/d and 120 s/d among the 1.0 W/cm2,2.0 W/cm2 and 3.0 W/cm3 were significant(F=9.314,F=8.584,F=9.026,P<0.05),and the MSC absorbance levels of 1.0 W/cm2 at once,24 h and 48h post ultrasound radiation were significantly higher than that of 2.0 W/cm2 and 3.0 W/cm3,respectively.2)The differences of the expression levels of bax protein,bc1-2 protein and bc1-2/bax were significantly(F=9.333,F=7.962,F=29.514,P<0.05).The differences of the number of capillaries and the number of fluorescently labeled positive cells among the three groups were significant(F=25.043,F=36.697,P<0.05).3)At 24 hours after transplantation,the positive ratio of MSC myocardial homing of ultrasound microbubble+MSC group was the highest,and the difference of that among three groups was significant(F=166.411,P<0.05).Conclusion:Ultrasound radiation microbubbles can promote the transplantation of autologous bone marrow mesenchymal stem cells(BM-MSCs)in rats with Adriamycin cardiomyopathy,and promote myocardial angiogenesis,and improve the damaged cardiac function by Adriamycin myocardial infarction.