目的基于网络药理学方法分析临床治疗糖尿病肾病(DN)的有效方剂水陆二仙丹治疗糖尿病肾病的药理机制,为其临床运用和新药研发提供参考。方法结合文献,通过中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)获取芡实、金樱子的主要化学成分及其靶点,根据ADME筛选中药活性组分;通过GenCards、OMIM、TTD、DRUGBANK数据库获取糖尿病肾病主要靶点,利用STRING平台进行蛋白质相互作用分析,而后采用Cytoscape 3.8.0软件构建"中药-成分-疾病-靶点"网络。构建PPI网络并挖掘网络中潜在的蛋白质功能模块。采用R 3.6.0分析"药物-成分-靶点-疾病"及其参与的生物过程及通路。结果水陆二仙丹治疗糖尿病肾病的核心活性成分为β-谷甾醇、槲皮素、山柰酚、维生素E等,核心靶点有AKT1、MAPK1、PTGS2、JUN等,水陆二仙丹治疗糖尿病肾病生物学通路主要作用于AGE-RAGE信号通路、TNF通路、VEGF通路等,其功能主要为调节细胞内激素受体的生命活动等。结论本研究初步揭示了水陆二仙丹治疗糖尿病肾病的多成分、多靶点、多通路的作用机制,为水陆二仙丹后续研究和临床应用提供了理论基础。
Background Our previous studies have demonstrated that Tongxinluo （TXL）, a traditional Chinese medicine, can protect hearts against no-reflow and reperfusion injury in a protein kinase A （PKA）-dependent manner. The present study was to investigate whether the PKA-mediated cardioprotection of TXL against no-reflow and reperfusion injury relates to the inhibition of myocardial inflammation, edema, and apoptosis. Methods In a 90-minute ischemia and 3-hour reperfusion model, minipigs were randomly assigned to sham, control, TXL （0.05 g/kg, gavaged one hour prior to ischemia）, and TXL ＋ H-89 （a PKA inhibitor, intravenously and continuously infused at 1.0 μg/kg per minute） groups. Myocardial no-reflow, necrosis, edema, and apoptosis were determined by pathological and histological studies. Myocardial activity of PKA and myeloperoxidase was measured by colorimetric method. The expression of PKA, phosphorylated cAMP response element-binding protein （p-CREB） （Ser133）, tumor necrosis factor a （TNF-a）, P-selectin, apoptotic proteins, and aquaporins was detected by Western blotting analysis. Results TXL decreased the no-reflow area by 37.4% and reduced the infarct size by 27.0% （P〈0.05）. TXL pretreatment increased the PKA activity and the expression of Ser133 p-CREB in the reflow and no-reflow myocardium （P 〈0.05）. TXL inhibited the ischemia-reperfusion-induced elevation of myeloperoxidase activities and the expression of TNF-a and P-selectin, reduced myocardial edema in the left ventricle and the reflow and no-reflow areas and the expression of aquaporin-4, -8, and -9, and decreased myocytes apoptosis by regulation of apoptotic protein expression in the reflow and no-reflow myocardium. However, addition of the PKA inhibitor H-89 counteracted these beneficial effects of TXL. Conclusion PKA-mediated cardioprotection of TXL against no-reflow and reperfusion injury relates to the inhibition of myocardial inflammation, edema, and apoptosis in the reflow and no-reflow myocardium.
LI Xiang-dongYANG Yue-jinCHENG Yu-tongDOU Ke-feiTIAN YiMENG Xian-min
