从RAAS激活通路探讨壮药壮通饮干预冠心病心肌缺血血瘀模型大鼠的分子靶点

CHD(coronary heart disease,CHD)is a serious disease that threatens to people's health . Recently a number of researches have shown that oxidative stress injury is an important mechanism in the development and progression of syndrome of blood stasis in myocardial ischemia of CHD due to RAAS (renin-angiotensin-aldosterone system，RAAS) activation. We have drawn out the pattern of signal molecules in the path of oxidative stress injury due to RAAS activation in syndrome of blood stasis in our premilinary work, moreover, in our clinical research we have observed that Zhuang minority medicine Zhuang Tong Yin formulae has an actual effect in the treatment of syndrome of blood stasis by promoting blood circulation to remove blood stasis so we speculate that Zhuang Tong Yin formulae takes an effect by affecting the high specificity and sensibility targeted molecules . On the base of our work, we will set up rat model of syndrome of blood stasis of myocardial ischemia of CHD, and firstly detect signal molecules and proteins by applying RT-PCR,ELISA,Western-blot, immunohistochemics, flow cytometry technologies ,then filtrate the high specificity and sensibility molecules that are able to reflect the degree of oxidative stress injury by data mining , lastly under the interference of Zhuang Tong Yin formulae, survey and verify the targeted effects of Zhuang Tong Yin formulae on the high specificity and sensibility molecules by enlarging samples and charify the therapeutic mechanism of Zhuang Tong Yin formulae in the treatment of syndrome of blood stasis , meantime,our findings are expected to provide some ideas for the targeted therapy of CHD.

冠心病严重危害人类的健康，近年研究表明肾素－血管紧张素－醛固酮系统(renin-angiotensin-aldosterone system，RAAS)激活启动的氧化应激损伤是冠心病心肌缺血血瘀证发生发展的重要机制，我们在前期基础研究中描绘了氧化应激反应RAAS激活通路信号分子导致血瘀证的模式，在临床中观察到具有活血化瘀作用的壮药经验方壮通饮对血瘀证具有确切的疗效，我们推测壮通饮通过影响通路中高特异性、敏感性靶点分子而发挥疗效，本课题在此工作基础上，建立冠心病心肌缺血血瘀证大鼠模型，首先采用RT-PCR、ELISA、Western-blot、免疫组化、流式细胞等技术或方法检测信号分子或蛋白，然后通过数据挖掘，筛选反映氧化应激损伤程度的高特异性、敏感性分子，最后通过壮通饮干预，扩大样本观察和验证其对高特异性、敏感性分子的靶向作用，阐明其治疗机制，研究结果也为建立冠心病的靶向治疗策略提供思路。

冠心病是一种慢性进展性疾病，危害人类健康并给社会造成了沉重的医疗负担，因此对冠心病发生机制及治疗需要深入的研究。部分壮族医药对冠心病有确切的疗效，但其起效机制仍不明确。本项目通过动物实验，建立冠心病心肌缺血血瘀证大鼠模型，采用壮药经验方壮通饮干预，检测反映氧化应激损伤程度的高特异性、敏感性分子变化，从氧化应激损伤角度阐明冠心病发病及壮通饮的起效机制。研究结果表明壮通饮高剂量组超氧化物歧化酶水平（198.55±2.89）升高，谷胱甘肽过氧化物酶活力（181.90±5.13）升高，缝隙连接蛋白43蛋白表达（0.55±0.02）升高，缝隙连接蛋白43mRNA相对表达（0.42±0.10）升高，白细胞介素10（IL-10）表达水平（36.62±3.33）升高，丙二醛水平（8.70±0.39）下降，内皮素1浓度（120.97±6.65）下降，血管紧张素ⅡmRNA相对表达（2.01±0.29）下降，血管紧张素Ⅱ 1型受体蛋白表达（2.75±0.20）降低， 血管紧张素Ⅱ 1型受体mRNA相对表达（1.96±0.35）下降，肿瘤坏死因子α（TNF-α）表达水平（116.80±6.61）下降、白细胞介素6（IL-6）表达水平（82.75±3.45）下降，与模型组比较，差异均有统计学意义（P<0.05 或P<0.01）。研究结果提示肾素－血管紧张素－醛固酮系统（RAAS）激活启动的氧化应激损伤是冠心病心肌缺血血瘀证发生发展的重要机制，壮通饮通过提高机体抗氧化能力，抑制氧化应激发生，减轻氧化应激对心肌的损害；同时，缓解血管收缩，改善心肌缺血缺氧状况，而且，减少大鼠心肌缺血再灌注引起的再灌注损伤发生率，从而降低再灌注损伤，减轻炎症反应，对心肌起到保护作用，是壮通饮治疗冠心病心肌缺血血瘀证的起效机制。本项目研究为壮药经验方壮通饮治疗冠心病心肌缺血的临床应用提供了基础研究的依据。