基于“痰瘀阻络说”研究“搜剔络道痰瘀同治”对DCM大鼠心肌AGEs及所致病变的干预作用

Our previous studies had shown that the treatment of activate blood and removing blood stasis has a better effect on myocardial fibrosis and vascular disease in diabetic cardiomyopathy (DCM), and its correlated with the inhibition of oxidative stress, NF -κB signal pathway and the content of Ang Ⅱand TGF-β1/Smads. In general, DCM has a long course of disease and its pathological change had been according with the feature of "long illness into the collaterals". So, can we use "Picking evils in collaterals with insects drugs" to improve the efficacy? The glycosylation end products (AGEs), which had an important role in the development and progression of DCM, and it had the characteristics of "Phlegm", and their lesions of myocardial microvascular and ECM were associated with "Blood Collaterals" and "Gas Collaterals" respectively. Therefore the opinion were put forward in our project that the accumulation of AGEs and the myocardial fibrosis exist the pathogenesis of "phlegm and blood stasis combined each other with blocking collaterals" and the method of "Picking evils in collaterals and Treated with Phlegm and Blood Stasis" were used to treat. Then the prevention and treatment effect of DCM will be observed by the method of taking the combination decoction of Didang and Xiao Xianxiong. The mechanism of it was explored from the accumulation of AGEs, AGEs-RAGE axis of the series reaction and degradation of AGEs and the other different segments. Our project results will contribute to understand the scientific connotation of "Stagnation of phlegm and blood stasis" and "Picking evils in collaterals and Treated with Phlegm and Blood Stasis" together and to find some effective methods and prescription for the prevention and control of DCM. What more, it can offer the foundation for the development and application of the classical prescription.

前期研究发现，活血化瘀法对糖尿病心肌病（DCM）中的心肌纤维化和血管病变具有较好防治作用，机制与抑制氧化应激、心肌组织AngⅡ的含量、TGF-β1/Smads以及NF-κB信号通路有关。DCM病理改变符合“久病入络”特征，一般的活血化瘀效果不佳，能否使用虫蚁之类“搜剔络道”来提高疗效？在该病的发生发展中起重要作用的糖基化终末产物（AGEs）本身具有“痰”的特征，其所导致的病变——心肌微血管和ECM分别与“血络”、“气络”吻合，故本课题提出AGEs的蓄积及引发的心肌病变存在“痰瘀互结阻络”病机，采用“搜剔络道痰瘀同治”法，以抵当汤与小陷胸汤合方为代表，观察该治法对DCM防治作用。从AGEs的蓄积、AGEs-RAGE轴的系列反应和AGEs的降解等不同环节探索其作用机制。以冀部分揭示“痰瘀阻络”和“搜剔络道、痰瘀同治”法的科学内涵，同时为防治DCM寻找有效的治法和方药，为经方的开发应用奠定基础。

糖尿病心肌病（DCM）病理改变符合“久病入络”特征，其病程中起重要作用的糖基化终末产物（AGEs）蓄积及引发的心肌病变存在“痰瘀互结阻络”病机。本项目主要研究采用“搜剔络道，痰瘀同治”法，通过观察该治法对AGEs的蓄积、AGEs-RAGE轴的系列反应和AGEs的降解等作用，探讨其防治DCM的作用机制。主要研究内容与结果如下：.①通过观察DCM模型大鼠心肌超微结构和功能的变化，发现“搜剔络道，痰瘀同治”法对心肌组织具有保护作用，能减缓糖尿病心肌病变的发生，减轻病变程度。.②运用酶联免疫吸附法、Western blotting及PCR技术检测心肌组织AGEs、RAGE及相关蛋白在DCM模型大鼠体内的表达，证实“搜剔络道，痰瘀同治”法干预糖尿病心肌病变的机制，与减少AGEs蓄积、加速AGEs降解有关。.③“搜剔络道，痰瘀同治”之抵当陷胸汤含药血清作用于心肌微血管内皮细胞，能抑制AGEs-RAGE轴的表达，降低活性氧浓度，证实该治法能抑制AGEs-RAGE轴的过度激活及抑制氧化应激反应，减轻心肌微血管内皮细胞损伤。.④“搜剔络道，痰瘀同治”法可降低DCM模型大鼠心肌组织炎症因子的表达、改善心肌纤维化病变，证实糖尿病心肌病变与AGEs-RAGE轴激活引发的系列反应有关。.⑤开展抵当陷胸汤HPLC-MS指纹图谱检测。.本研究揭示了“搜剔络道、痰瘀同治”法防治DCM的部分作用机制，阐明了“痰瘀互结阻络”的部分科学内涵，为临床防治DCM提供了新的治疗思路，也为经方的开发应用奠定了基础。