电针诱导骨髓间充质干细胞分化为血管生长因子的机制研究  

Mechanism of Electro-Acupuncture Inducing Differentiation of BMSCs into Vascular Growth Factors

作　　者：王秀珍[1,2] 贾坤平[1,2] 赵楠 倪虹[1,2] 王虹[1,2] 王亚楠 王树人[1,3] 彭宇飞 吴全娥[4] WANG Xiuzhen;JIA Kunping;ZHAO Nan;NI Hong;WANG Hong;WANG Yanan;WANG Shuren;PENG Yufei;WU Quan'e(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;Hebei University of Chinese Medicine, Shijiazhuang 050200, 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 [4]河北中医学院,河北石家庄050200

出　　处：《针灸临床杂志》2021年第8期74-81,共8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:探讨电针诱导骨髓间充质干细胞(BMSCs)分化为血管生长因子的机制研究。方法:全骨髓贴壁法分离、传代BMSCs;流式细胞仪鉴定BMSCs表面抗原CD29。将SD大鼠随机分为假手术组、模型组、电针组、BMSCs组和电针+BMSCs组,制备MIRI模型。假手术组只结扎不穿线;模型组、电针组采用心肌内直接注射生理盐水的方法;BMSCs组、电针+BMSCs组将BMSCs移植到梗死边缘区;分别在移植后3 d、7 d和14 d共3个时间点电镜下观察心肌细胞超微结构改变,免疫组化法检测心肌组织VEGF、TGF-β1蛋白含量的变化;Western blot法检测Ang-1、bFGF蛋白含量。结果:CD29阳性率高于90%;与假手术组比较,模型组心肌线粒体肿胀明显,VEGF、TGF-β1、Ang-1和bFGF蛋白水平明显升高,差异具有统计学意义(P<0.05);与模型组比较,各治疗组心肌线粒体肿胀明显缓解,VEGF、Ang-1和bFGF蛋白水平明显升高,差异具有统计学意义(P<0.05),其中以14 d时电针+BMSCs组升高显著,差异具有统计学意义(P<0.05);与再灌注3 d、7 d比较,14 d时电针+BMSCs组TGF-β1蛋白水平明显降低,差异具有统计学意义(P<0.05)。结论:电针能够诱导BMSCs分化为血管生长因子,其机制可能与电针促进BMSCs在心肌内生长增殖,明显改善MIRI心肌组织超微细胞结构损伤,上调VEGF、Ang-1和bFGF蛋白水平,动态调控TGF-β1水平有关。Objective:To investigate the mechanism of electro-acupuncture inducing differentiation of bone mesenchymal stem cells(BMSCs)into vascular growth factors.Methods:The whole bone marrow adherence method was used for the isolation and passage of rat BMSCs,and flow cytometry was applied for the identification of BMSCs surface antigen CD29.SD rats were randomly divided into the sham operation group,the model group,the electro-acupuncture group,the BMSCs group and the combined group(electro-acupuncture+BMSCs).Ligation was performed without threading in the sham operation group.After successful modeling of myocardial ischemia reperfusion injury(MIRI),the model group and the electro-acupuncture group were injected with normal saline directly into the myocardium;in the BMSCs group and the combined group,BMSCs were transplanted to the marginal area of infarction;the ultrafine structure changes of myocardium were observed 3 d,7 d and 14 d after the transplantation under electron microscope;immunohistochemical method was used to detect the protein contents of VEGF and TGF-β1 in myocardial tissue;Western blot method was used to detect the protein contents of Ang-1 and bFGF.Results:The positive rate of CD29 was higher than 90%.Compared to those in the sham operation group,the swelling of myocardial mitochondria was obvious in the model group;the protein levels of VEGF,TGF-β1,Ang-1 and bFGF increased significantly(P<0.05).Compared to those in the model group,the swelling of myocardial mitochondria in each treatment group was significantly relieved;the protein levels of VEGF,Ang-1 and bFGF were significantly increased(P<0.05),of which the increases in the combined group were more significant on 14 d(P<0.05);Compared to that after 3 d and 7 d of reperfusion,the protein level of TGF-β1 was significantly decreased in the combined group after 14 d of reperfusion(P<0.05).Conclusion:Electro-acupuncture can induce BMSCs to differentiate into vascular growth factors.The mechanism may be related to promoting the growth and proliferatio

关 键 词：电针 骨髓间充质干细胞 心肌缺血再灌注损伤 血管生长因子 血管修复 

分 类 号：R245.97[医药卫生—针灸推拿学]