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银茶复方抑制实验性兔动脉粥样硬化的药效学研究

Study of the Anti-atherosclerotic Effect of Yincha Compound in Rabbits with Experimental Atherosclerosis

【作者】 姚祥鑫

【导师】 蒋琳兰;

【作者基本信息】 华南理工大学 , 发酵工程, 2010, 硕士

【摘要】 目的:在成功建立兔动脉粥样硬化模型的基础上,通过观察实验兔血脂水平、动脉粥样硬化斑块的面积、主动脉核转录因子、金属基质蛋白酶-2表达水平的变化等,研究银茶复方抗动脉粥样硬化的药效学作用及探讨其作用机制,比较了银茶复方及其组方成分茶多酚与银杏叶提取物对动脉粥样硬化作用的差异,为研究该复方制剂的物质基础和临床治疗提供理论依据。方法:雄性新西兰白兔35只随机均分为7组,分别为:正常对照组、模型组、银茶复方高剂量组、银茶复方低剂量组、辛伐他汀组、茶多酚组和银杏叶提取物组。正常对照组喂食普通饲料,其它组喂食高脂饲料,银茶复方高剂量组、银茶复方低剂量组、辛伐他汀组、茶多酚组和银杏叶提取物组同时每天灌胃银茶复方、银茶复方、辛伐他汀、茶多酚、银杏叶提取物各500mg/﹒kg-1﹒d-1、200mg﹒kg-1﹒d-1、5mg﹒kg-1﹒d-1、250mg﹒kg-1﹒d-1、250mg﹒kg-1﹒d-1,共喂养10周。于实验前、实验结束时检测血清TC、TG、LDL-C、HDLC、MDA、SOD并计算动脉粥样硬化指数;HE染色和MASSON染色观察主动脉组织形态学变化;图像分析软件测定主动脉内膜厚度、中膜厚度、内膜厚度/中膜厚度及粥样斑块面积百分比;免疫组织化学法检测主动脉粥样硬化斑块中NF-ΚB、MMP-2的蛋白表达,RT-PCR检测主动脉粥样硬化斑块中MMP-2的mRNA表达。结果:实验结束时,除对照组外的其它各组TC、TG、LDL-C、HDLC、MDA、SOD显著增高,主动脉见典型脂质条纹,局部斑块形成,病变处NF-ΚB、MMP-2显著表达。与模型组相比,①银茶复方高剂量组、银茶复方低剂量组血脂水平改善,内膜厚度显著减轻,粥样斑块面积减少,NF-ΚB、MMP-2的表达减少(P<0.05),银茶复方高剂量组与银茶复方低剂量组相比上述改善更为明显(P<0.05);②茶多酚组、银杏叶提取物组血脂水平改善,内膜厚度显著减轻,粥样斑块面积减少,减少NF-ΚB、MMP-2的表达(P<0.05),茶多酚组较银杏叶提取物组作用更为明显(P<0.05),以茶多酚与银杏叶提取物联合的银茶复方疗效更加显著(P<0.05)。结论:银茶复方可通过调节血脂水平、抑制氧化反应、稳定斑块、抑制体内局部炎症反应达到抑制动脉粥样硬化病变的作用;银茶复方对动脉粥样硬化的抑制作用有一定的剂量效应;银茶复方中对抑制动脉粥样硬化起主要作用的单方是茶多酚,银杏叶提取物起辅助性作用。

【Abstract】 Objectives:To investigate the ability of anti-atherosclerosis of Yincha compound by observe changes the serum lipid levels, the area of plaque,the expression of nuclear factor kappaB and matrix metalloproteinase-2,and then to ascertain its mechanisms,and compare the effect of tea polyphenols and inko biloba extract so as to provide the therapeutic basis to material research and clinical coronary disease.Methods:Thirty five male New Zealand rabbits were randomly allocated into five equal groups:NC group;AS group;YCH group;YCL group;Sim group;TP group and GBE group.The NC group were fed with basis forage,the rest were fed with high-fat gorage.The NC group,AS group,YCH group,YCL group,Sim group,TP group and GBE group were independently given Yincha compound, Yincha compound, Simvastatin, tea polyphenols,ginko biloba extract 500mg/﹒kg-1﹒d-1、200mg﹒kg-1﹒d-1、5mg﹒kg-1﹒d-1、250mg﹒kg-1﹒d-1、250mg﹒kg-1﹒d-1 , They were fed for 10 weeks in all.Serum levels were examined before experiment and at the end of 10 week and AI were calculated. The histomorphology of the aorta was observed by HE stain and MASSON stain.The IT,MT,I/M and atherosclerotic area ratio were measured by image analysis software. The protein expression of NF-κB,MMP-2were detected by immunohistochemitry staining.The mRNA levels of MMP-2 were detected by RT-PCR.Results:At the end of 10 weeks, In all high cholesterol-fed rabbits,serum lipid level increased remarkly, The typical fatty streaks and early atheromatous Plaque on the aorta have been formed and the expression of NF-ΚB and MMP-2 in the lesion were inereased significantly. Compare to atherosclerosis group:①the thickeness of intima and media,and the area of atheromatous Plaque were lower in YCH group and YCL group,especially in YCH group(P<0.05);②the thickeness of intima and media,and the area of atheromatous Plaque were lower in TP group and GBE group,especially in TP group,and the comnination of both of them gain a best effect(P<0.05).Conclusions:Yincha compound prevent the development of atherosclerosis through lowering lipid level, oxidizing reaction,stabilizing plaque and inhibiting inflammatory reaction.

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