丹参通络解毒汤联合内皮祖细胞移植对心肌缺血再灌注损伤大鼠促血管新生作用的研究  

Study on Effect of Danshen Tongluo Jiedu Decoction(丹参通络解毒汤) Combined with Endothelial Progenitor Cells Transplantation on Promoting Angiogenesis in Rats with Myocardial Ischemia Reperfusion Injury

作　　者：李彩云 肖青 李鑫辉 杜建芳 王静雯 陈欣 IJ Caiyun;XIAO Qing;LI Xinhui;DU Jianfang;WANG Jingwen;CHEN Xin(TCM College of Hunan University of Chinese Medicine,Changsha 410208,Hunan,China)

机构地区：[1]湖南中医药大学中医学院,湖南长沙410208

出　　处：《中华中医药学刊》2022年第1期25-29,I0014-I0016,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074392);湖南省自然科学基金面上项目(2019JJ40210)。

摘　　要：目的探讨丹参通络解毒汤(DSTLJD)联合内皮祖细胞(endothelial progenitor cells, EPCs)移植对心肌缺血再灌注损伤(ischemic reperfusion injury, IRI)模型大鼠促血管新生作用的影响及其机制。方法采用密度梯度离心法及差数贴壁法分离和培养EPCs,采用免疫荧光法鉴定EPCs。通过结扎左冠状动脉左前降支的方法建立IRI模型,采用随机数字表法分为SH组(假手术组)、IRI组(模型组)、EPCs组(EPCs移植组)、DSTLJD组(丹参通络解毒汤组)、DSTLJD+EPCs组(丹参通络解毒汤+EPCs移植),每组10只。4周后,采用HE染色法观察心肌组织的病理形态,用免疫组化法测定各组大鼠心肌微血管计数(microvessel count, MVC)、心肌微血管密度(microvessel density, MVD),并检测各组大鼠血管内皮生长因子(vascular endothelial growth factor, VEGF)、碱性成纤维生长因子(basic fibroblast growth factor, bFGF)的蛋白表达。结果与IRI组比较,EPCs组、DSTLJD组、DSTLJD+EPCs组大鼠细胞肿胀及坏死程度减轻,MVC、MVD及VEGF、bFGF蛋白表达显著增高(P<0.01);与EPCs组相比,DSTLJD组、DSTLJD+EPCs组大鼠炎性细胞浸润及细胞肿胀程度轻微,MVC、MVD及VEGF、bFGF蛋白表达增高(P<0.05,P<0.01);与DSTLJD组相比,DSTLJD+EPCs组大鼠肌束排列较整齐,无明显细胞坏死,MVC、MVD及VEGF、bFGF蛋白表达明显增高(P<0.01)。结论丹参通络解毒汤联合EPCs移植可能通过上调VEGF、bFGF的表达,促进心肌IRI大鼠的血管新生,从而保护IRI心肌。Objective To investigate the effect and mechanism of Danshen Tongluo Jiedu Decoction(丹参通络解毒汤,DSTLJD)combined with endothelial progenitor cells(EPCs)transplantation on promoting angiogenesis in rats with myocardial ischemia reperfusion injury(IRI).Methods The density gradient centrifugation and differential adherence methods were used to isolate and culture EPCs,identificating the EPCs by immunofluorescence.The IRI model was established by ligating the left anterior descending coronary artery.The rats were grouped by means of random digital tables:sham operation group,IRI group,EPCs transplantation group,DSTLJD group,DSTLJD combined with EPCs transplantation group,10 rats in each group.After 4 weeks,the pathological morphology of myocardium was observed by hematoxylin eosin staining.Myocardial microvessel count(MVC)and myocardial microvessel density(MVD)in rats of each group were measured by immunohistochemical method.The expressions of vascular endothelial growth factor(VEGF)and basic fibroblast growth factor(bFGF)in each group were detected.Results Compared with those of IRI group,the swelling and necrosis of the cells in EPCs transplantation group,DSTLJD group,DSTLJD combined with EPCs transplantation group were reduced,meanwhile levels of MVC and MVD as well as protein expressions of VEGF and bFGF were significantly increased.The difference was statistically significant(P<0.01).Compared with those of EPCs transplantation group,the infiltration of inflammatory cells and the degree of cell swelling in DSTLJD group and DSTLJD combined with EPCs transplantation group were slight,meanwhile levels of MVC and MVD as well as protein expressions of VEGF and bFGF were increased(P<0.05,P<0.01).Compared with those of DSTLJD group,the muscle bundles in DSTLJD combined with EPCs transplantation group were arranged in good order and there was no obvious cell necrosis.Meanwhile,levels of MVC and MVD as well as protein expressions of VEGF and bFGF were significantly increased(P<0.01).Conclusion DSTLJD combined wi

关 键 词：丹参通络解毒汤 心肌缺血再灌注损伤 血管内皮生长因子 碱性成纤维生长因子 血管新生 

分 类 号：R285.5[医药卫生—中药学]