化痰化瘀通络法对糖尿病大鼠心肌微血管和细胞外间质的影响及机制研究  

Effect and Mechanism of Resolving Phlegm,Dispersing Blood Stasis and Dredging Collaterals on Myocardial Microvasculature and Extracellular Matrix in Diabetic Rats

作　　者：蔡正银[1,2] 储全根[1,2] 储俊[2,3] 轩云 程捷 CAI Zheng-yin;CHU Quan-gen;CHU Jun;XUAN Yun;CHENG Jie(College of Traditional Chinese Medicine,Anhui University of Traditional Chinese Medicine,Hefei,230038;Key Laboratory of Xin'an Medical Science,Ministry of Education,Anhui University of Chinese Medicine,Hefei,230038;Research and Experimental Center of Anhui University of Chinese Medicine,Hefei,230038)

机构地区：[1]安徽中医药大学中医学院,合肥230038 [2]安徽中医药大学新安医学教育部重点实验室,合肥230038 [3]安徽中医药大学科研实验中心,合肥230038

出　　处：《中国中西医结合杂志》2020年第8期948-954,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81774189);安徽省教育厅优秀拔尖人才培育项目(No.gxgwfx2019027);安徽省科技攻关项目(No.1704f0804032)。

摘　　要：目的观察化痰化瘀通络法对糖尿病大鼠心肌微血管和细胞外间质的影响并探索部分机制。方法将糖尿病成功造模的40只清洁级SD大鼠随机分为:模型组(DCM组)、糖基化终末产物特异性蛋白交联抑制剂(ALT-711)组、化痰化瘀组(RPDBS组)、化痰组(RP组)、化瘀组(DBS组),每组8只,另选10只正常SD大鼠作对照组(NC组)。3周后各组开始灌胃相应药物8周,麻醉状态下取材。光镜观察心肌组织的形态变化和细胞外间质胶原的沉积情况,透射电镜观察心肌微血管的超微结构;免疫组织化学法检测心肌组织中Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(ColⅢ)和基质金属蛋白酶1(MMP-1)、基质金属蛋白酶组织抑制因子1(TIMP-1)蛋白的表达。结果光镜显示,与DCM组比较,各治疗组心肌细胞变性坏死程度情况改善,心肌胶原纤维增生减少,排列基本均匀,RPDBS组治疗效果优于RP组和DBS组;透射电镜结果显示,与DCM组比较,各治疗组心肌微血管内皮细胞和基底膜的破坏程度均改善,且RPDBS组改善情况优于RP组和DBS组;免疫组织化学法显示,与NC组比较,DCM组ColⅠ、ColⅢ、MMP-1、TIMP-1蛋白表达升高(P<0.01,P<0.05);与DCM组比较,各治疗组均下调了ColⅠ、ColⅢ和MMP-1、TIMP-1蛋白表达水平(P<0.01,P<0.05),其中RPDBS组较RP组和DBS组蛋白表达水平下调明显(P<0.01)。结论化痰化瘀通络法能够改善糖尿病大鼠心肌微血管及细胞外间质病变,其机制可能与下调心肌组织MMP-1、TIMP-1蛋白表达水平,调节MMP-1/TIMP-1平衡,改善心肌细胞外基质代谢有关。且化痰化瘀通络法合用疗效优于化痰法和化瘀法单独应用。Objective To explore the effects and mechanism of resolving phlegm,dispersing blood stasis and dredging collaterals(RPDBS)on myocardial microvasculature and extracellular matrix in diabetic rats.Methods 40 clean SD diabetic rats were randomly divided into 4 groups:diabetic cardiomyopathy(DCM)group,Advanced glycosylation end products(AGEs)specific protein cross-linking inhibitor(ALT-711)group,RPDBS group,resolving phlegm(RP)group and dispersing blood stasis(DBS)group,8 rats in each group,and 10 normal SD rats were selected as normal control(NC)group.After 3 weeks,the rats of each group were disposed by intragastric administration of corresponding drugs for 8 weeks,and the samples were taken under anesthesia.The morphological changes of myocardial tissue and the deposition of collagen in extracellular matrix were observed under light microscopy,the ultrastructure of myocardial microvessels was surveyed under transmission electron microscopy,and the expressions of collagenⅠ(ColⅠ),collagenⅢ(ColⅢ),matrix metalloproteinase 1(MMP-1)and tissue inhibitor of metalloproteinases 1(TIMP-1)proteins in myocardial tissue were detected by immunohistochemistry.Results Compared with DCM group,the degree of degeneration and necrosis of cardiac myocytes in each treatment group was improved,the proliferation of collagen fibers in myocardium was reduced and arranged uniformly.The therapeutic effect of RPDBS group was better than that of RP group and DBS group.The results of transmission electron microscopy showed that the degree of destruction of myocardial microvascular endothelial cells and basement membrane in each treatment group was higher than that of DCM group.The improvement of RPDBS group was better than that of RP group and DBS group.Compared with NC group,the expression levels of ColⅠ,ColⅢ,MMP-1 and TIMP-1 were increased in DCM group(P<0.01,P<0.05).Compared with DCM group,the expression levels of ColⅠ,ColⅢ,MMP-1 and TIMP-1 were decreased in all treatment groups(P<0.01,P<0.05).The expression levels of ColⅠ,C

关 键 词：化痰化瘀通络 心肌微血管 细胞外间质 机制 

分 类 号：R285.5[医药卫生—中药学]