电针联合骨髓间充质干细胞促进心肌缺血再灌注损伤大鼠血管新生作用机制研究  

Mechanism of Electro-Acupuncture Combined with BMSCs on Angiogenesis in Rats with MIRI

作　　者：王秀珍[1,2] 邹伟[1,3] 马育轩[1] 赵楠[1] 高长玉[1] 王特哈斯[1] 彭宇飞 WANG Xiuzhen;ZOU Wei;MA Yuxuan;ZHAO Nan;GAO Changyu;WANG Tehasi;PENG Yufei(Heilongjiang University of Chinese Medicine, Harbin 150040, China;The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150001, China;The First Affiliated Hospital of Heilongjiang University of Chinese Medicine, Harbin 150040, China)

机构地区：[1]黑龙江中医药大学,黑龙江哈尔滨150040 [2]黑龙江中医药大学附属第二医院,黑龙江哈尔滨150001 [3]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《针灸临床杂志》2021年第5期68-76,共9页Journal of Clinical Acupuncture and Moxibustion

基　　金：国家自然科学基金面上项目,编号:81874426;黑龙江省博士后科研基金面上项目,编号:LBH-Z16194;黑龙江省中医药管理局科技计划项目,编号:ZHY2020-116;黑龙江中医药大学博士创新基金,编号:2019BS10;黑龙江中医药大学科研基金面上项目,编号:2019MS37;黑龙江中医药大学生创新创业训练计划项目,编号:201910228075。

摘　　要：目的:探讨电针联合BMSCs促进心肌缺血再灌注损伤模型大鼠血管新生的作用机制研究。方法:倒置显微镜下观察细胞生长形态;MTT法检测细胞增殖情况;流式细胞仪鉴定BMSCs表面抗原。将SD大鼠随机分为5组,分别为假手术组、模型组、电针组、BMSCs组和电针+BMSCs组,制备MIRI模型。模型组、电针组直接注射生理盐水;BMSCs组、电针+BMSCs组将BMSCs移植到梗死边缘区。在3 d、7 d和14 d检测CK-MB、cTnI含量;TTC染色测定梗死面积;光镜下观察病理形态学改变;荧光标记观察BMSCs增殖情况;免疫组化法检测心肌组织CD31、SDF-1蛋白表达。结果:细胞增殖较为活跃,生长曲线呈S形,表面抗原CD45阳性率<5%,CD90阳性率>90%。再灌注后,随着时间的延长,电针+BMSCs组能够明显降低心肌酶及心肌梗死面积的水平(P<0.05);病理形态学显示电针+BMSCs移植组能够减少炎性细胞浸润;荧光标记显示电针+BMSCs组能够明显促进BMSCs在心肌内生长增殖;电针+BMSCs组CD31、SDF-1蛋白表达明显优于其他治疗组(P<0.05),其中14 d时疗效最佳。结论:随着治疗时间的延长电针诱导BMSCs移植后对心肌具有明显的保护作用,其机制可能与电针促进BMSCs在心肌内生长增殖、有效减少心肌损伤、减小心肌梗死面积、改善MIRI心肌组织病理形态、增加微血管密度和促进缺血区边缘血管新生有关。Objective:To explore the mechanism of electro-acupuncture combined with bone marrow mesenchymal stem cells(BMSCs)on angiogenesis in rats with myocardial ischemia-reperfusion injury(MIRI).Methods:The cell growth morphology was observed under inverted microscope,the proliferation of cells was detected by MTT method,and BMSCs surface antigen was identified by flow cytometry.SD rats were randomly divided into five groups,which were the sham operation group,the model group,the electro-acupuncture group,the BMSCs group,as well as the electro-acupuncture and BMSCs group.The model group and the electro-acupuncture group were injected with normal saline directly;BMSCs were transplanted to the border area of infarction in the BMSCs group and in the electro-acupuncture and BMSCs group.The contents of CK-MB and cTnI were detected after 3 d,7 d and 14 d.Infarct area was measured by TTC staining,pathomorphological changes were observed under light microscope,proliferation of BMSCs was observed by fluorescent labeling,and the protein expressions of CD31 and SDF-1 in myocardial tissue were detected by immunohistochemistry.Results:The cell proliferation was active,the growth curve was S-shaped,the positive rate of CD45 was less than 5%,and the positive rate of CD90 was more than 90%.After reperfusion,the level of myocardial enzyme and the area of myocardial infarction were significantly reduced in the electro-acupuncture and BMSCs group(P<0.05).The pathological morphology showed that the intervention of electro-acupuncture and BMSCs could reduce inflammatory cell infiltration;the fluorescence markers showed that the intervention of electro-acupuncture and BMSCs could promote the growth and proliferation of BMSCs in the myocardium;the expressions of CD31 and SDF-1 in the electro-acupuncture and BMSCs group were significantly improved than those of the other treatment groups(P<0.05),the best effect was on 14th day.Conclusion:With the prolongation of treatment,the BMSCs implant induced by electro-acupuncture has significant protecti

关 键 词：心肌缺血再灌注 电针 骨髓间充质干细胞 血管新生 

分 类 号：R246.1[医药卫生—针灸推拿学]