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目的:观察血府逐瘀汤复方对抗糖尿病血瘀证患者血清致血管内皮功能障碍的作用机制。方法:ECV-304分组后以MTT法选取该复方药物血清的最佳作用浓度,检测ET和NO、内皮细胞蛋白C受体(EPCR)、血管内假性血友病因子(vWF)和血栓调节蛋白(sTM)的含量。在激光扫描共聚焦显微镜下观察其对细胞内Ca2+浓度([Ca2+]i)和细胞骨架微丝结构的影响。结果:与模型组相比,药物组细胞活性和([Ca2+]i)浓度升高,ET/NO改变,EPCR、vWF和sTM降低,骨架微丝减少。结论:该复方可以有效对抗糖尿病血瘀证患者血清干预ECV-304造成的内皮功能障碍,从药物的角度反证了细胞损伤模型的建立。
Objective: To observe the mechanism of compound Xuefu Zhuyu decoction against vascular endothelial dysfunction caused by diabetes mellitus patients with blood stasis syndrome. METHODS: After ECV-304 grouping, the optimal concentration of the compound drug serum was selected by MTT assay to detect ET and NO, endothelial cell protein C receptor (EPCR), intravascular von Willebrand factor (vWF) and thrombus regulation. The content of protein (sTM). The effects of intracellular Ca2+ concentration ([Ca2+]i) and cytoskeletal microfilament structure were observed under a laser scanning confocal microscope. RESULTS: Compared with the model group, the cell activity and ([Ca2+]i) concentrations in the drug group increased, ET/NO changed, EPCR, vWF, and sTM decreased, and the number of microfilaments in the skeleton decreased. Conclusion: The compound can effectively counteract the endothelial dysfunction caused by serum of patients with diabetes blood stasis syndrome by intervention of ECV-304, and the establishment of cell injury model is reconfirmed from the perspective of drugs.