【作者】 孙静；

【导师】 周亚滨；

【作者基本信息】 黑龙江中医药大学 ， 中医内科学， 2008， 博士

【摘要】 目的:通过观察以益气养心安神为立法组成的中药复方养心颗粒对近似不稳定型心绞痛家兔模型颈总动脉血小板源生长因子-BmRNA及血管内皮生长因子mRNA表达的影响,从分子生物学水平探讨其可能的作用机制,为临床应用提供理论依据。方法:以球囊一次性损伤家兔颈动脉结合高脂饮食喂养,8周末后随机分为模型对照组、养心颗粒组,消心痛组和阿托伐他汀组,每组各7只,另取7只家兔为空白对照组,于10周末进行药物触发的方法复制成近似不稳定型心绞痛动物模型。运用体表彩色超声方法观察各组家兔颈总动脉内中膜厚度及血流变化情况:采用HE染色观察模型血管病理形态学变化;采用电镜观察超微结构变化:采用酶标法观察模型血脂成分含量变化;采用速率散射光比浊法测定血浆纤维蛋白原的水平;采用放免法检测血浆内皮素和一氧化氮含量的变化;采用RT-PCR法观察颈总动脉血小板源生长因子-BmRNA和血管内皮生长因子mRNA表达的变化。结果:1.根据体表超声测定,与空白对照组相比,各组在8周末时,均出现颈总动脉IMT明显增厚(P＜0.01),PI及RI显著增加(P＜0.05),而VS加快不明显,差异无统计学意义(P＞0.05)。10周末时,模型对照组IMT、PI、RI值继续增加,养心颗粒治疗组和消心痛治疗组的IMT、PI、RI值明显下降,与模型对照组相比,差异有统计学意义(P＜0.0 5),但不如阿托伐他汀组。2.病理研究结果表明,除空白对照组外,各组均出现血管内皮损伤,血管内膜增厚,脂肪沉积明显,脂质斑块形成。各治疗组颈总动脉内皮细胞轻度减少,内膜轻度增厚,内膜下脂质沉积减少。3.在第8周末时,除空白对照组外,各组TC、TG、LDL-C明显升高,HDL-C明显降低,差异有统计学意义(P＜0.01);第10周末时,模型对照组TG继续升高,与8周相比,差异有统计学意义(P＜0.01),TC、HDL-C、LDL-C均无明显变化(P＞0.05);养心颗粒组中的TG含量与8周末相比,无显著变化(P＞0.05),CHO和LDL-C明显降低,与8周末相比,差异有统计学意义(P＜0.05);阿托伐他汀组中TC、TG和LDL-C含量均有所降低,HDL-C升高,与8周末相比,差异有统计学意义(P＜0.05):消心痛治疗组各项指标均无明显影响(P＞0.05)。4.与空白对照组相比,各组在8周末Fbg的含量明显升高,差异有统计学意义(P＜0.01);10周末,与模型对照组相比,各治疗组Fbg含量均有所降低,差异有统计学意义(P＜0.01);各治疗组间比较,养心颗粒疗效优于其他两组,差异有统计学意义(P＜0.05)。5.与空白对照组相比,8周末各组血浆NO含量均显著降低,而血浆ET含量显著升高,差异有统计学意义(P＜0.01);10周末,与模型对照组比较,各治疗组血浆NO含量均有显著升高,血浆ET含量显著下降,差异有统计学意义(P＜0.05);各治疗组间相比较,消心痛组疗效优于另外两组,差异有统计学意义(P＜0.01),而养心颗粒与阿托伐他汀组相比,差异无统计学意义。6.与模型对照组相比,各治疗组均可下调PDGF-B mRNA表达(P＜0.01),养心颗粒治疗组的PDGF-B mRNA表达显著低于消心痛和阿托伐他汀治疗组(P＜0.05);同时各治疗组均可上调VEGF mRNA的表达(P＜0.01),养心颗粒治疗组的PDGF-B mRNA表达显著高于另外两组,差异有统计学意义(P＜0.05)。结论:以益气养心安神法组成的中药复方制剂养心颗粒对近似不稳定型心绞痛模型具有明确的防治作用,它可以下调PDGF-B mRNA表达,上调VEGF mRNA表达,其可能是通过调整人体阴阳平衡、改善血管内皮系统功能障碍、抑制血管内膜增生,从而改善模型血管的病理形态学的异常,减轻血管内膜增厚程度和斑块易损程度,起到防治不稳定型心绞痛的作用。更多还原

【Abstract】 Objective:To observer the effects of recuperating-heart granule that it can to tonify Qi and stabilize the heart and calm the mind on rabbit model of similar unstable angina that the expression of arteria carotis communis PDGF-B mRNA and VEGF mRNA,to investigate its mechanism of action at level of molecular biology,to provide a reliable theoretic basis for this therapy.Methods:Aortic wall injuries were introduced using an intravascular balloon in rabbits before they were fed on an a high fat diet for 8 weeks.At the end of week 8,rabbits were randomly divided into five groups.There are model control group; recuperating-heart granule group;Cedocard group;atorvastation group and blank group.There are 7 rabbits in every group.At the end of 10 weeks,to copy the model of similar unstable angina that used the method of Pharmacological triggering.to investigate the thickness and blood flow rate of arteria carotis communis by body surface chromatic color ultrasonography and to observe pathomorphism change of blood vessel by HE dyeing.to survey ultrastructure change by Electron microscope,and blood fat content change of model by enzyme linked immunosorbent assay;to determine fibrinogenic level by speed stray light turbidity method; detecting content change of blood serum NO and ET by immunology; observer the expressing change of arteria carotis communis PDGF-B mRNA and VEGF mRNA by RT-PCR.Results:1.According to the result of body surface ultrasonography,compare with the blank group,every group emerge that arteria carotis communis IMT obviously thicken(p＜0.01),PI and RI obviously addition(p＜0.05),but VS accelerate in nubibus.It was not considered statistically significant,at the end of 8 weeks.10 weeks later,model control group IMT PI RI continue increased,those compare with model control group,recuperating-heart granule group and Cedocard group IMT PI RI decreased obviously.It was considered statistically significant(p＜0.05),but the result in atorvastation group is better than them.2.Effect of pathology research indicated,besides black control group,every group all emerge that vascular endothelial cell damaged,endangium thick,fatty deposition obviously,fatty substance plaque formed,each treatment group,arteria carotis communis endothelial cell decrease lightly,endomembrane thick lightly,fatty deposition decrease under endomembrane.3.In the end of 8 weeks,TC TG LDL-C increase obviously and HDL-C decline obviously at each group except to blank control group.It was considered statistically significant(p＜0.01).TG at model group continue increased in ten weeks than 8 weeks,it was considered statistically significant(p＜0.01),TC、HDL-C、LDL-C has no any change(p＞0.05);TG have no significant change in Recuperating-heart granule group(p＞0.05),CHO and LDL-C obviously decline.it was considered statistically significant (p＜0.05).TC、TG and LDL-C decreased,HDL-C increased in atorvastation group,it was considered statistically significant (p＜0.05);the index has no any influence in Cedocard group.4.Compare with blank group,Fbg obviously increase in each group at the end of 8 weeks,it was considered statistically significant (p＜0.01);AT the end of 10 weeks,Fbg decline in treatment group to compare with control group it was considered statistically significant(p＜0.01),compare with each other during treatment group,the function of Recuperating-heart granule is better than other group,itwas considered statistically significant(p＜0.05). 5.At the end of 8 weeks,to compare with black group,the contents of blood plasm NO all reduced obviously,while the contents of blood serum ET increase obviously in model group,it was considered statistically significant(p＜0.01).At the end of 10 weeks,to compare with model control group,the contents of blood plasm NO all increased obviously,the contents of blood plasm ET decreased obviously in each treatment group,it was considered statistically significant.Compare with each other during treatment group,the function of Cedocard group is better than other two group,it was considered statistically significant.While there is similar function between Recuperating-heart granule group and atorvastation group,it was not considered statistically significant6.To compare with control group,the treatment group can down regulation expression of model PDGF-B mRNA.the express of Recuperating-heart granule group is lower than Cedocard group and atorvastation group(p＜0.05),at the same time,treatment group can up-regulation expression of VEGF mRNA(p＜0.01),the expression of PDGF-B mRNA in Recuperating-heart granule group significant higher than other two groups,it was considered statistically significant(p＜0.05).Conclusion:Recuperating-heart granule which can tonify Qi and stabilize the heart and calm the mind on the model of similar unstable angina has protective effect definitely.It can improve system functional disturbance of vessel endothelium,down regulation the expression of PDGF-B mRNA,up-regulation expression of VEGF mRNA.It may be to regulate yin-yang balance of human body to improve the functional disturbance of blood vessel endothelial system,to restrain endangium hyperplasia,thus it can improve pathology form and ultrastructure abnormality of model blood vessel,it also reduce the thickness of endangium and extent of easy damage plaque.It can prevention and cure unstable angina.更多还原