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目的研究九龙藤黄酮(Bauhinia championii flavones,BCF)抑制程序性坏死抗心肌缺氧/复氧损伤的作用及机制。方法建立心肌细胞缺氧/复氧损伤模型,给予九龙藤黄酮预处理,倒置显微镜观察心肌细胞形态学变化;以紫外分光光度法检测总抗氧化力(T-AOC);ELISA法测定肿瘤坏死因子(TNF-α)的含量;以Western-blot法检测受体相互作用蛋白激酶3(RIPK3)的蛋白表达;Annex v-FITC/PI双染法测定心肌细胞坏死率。结果与模型组相比,九龙藤黄酮预处理能减轻心肌细胞损伤,提高总抗氧化力,降低肿瘤坏死因子-α的含量,下调受体相互作用蛋白激酶3蛋白的表达,降低心肌细胞坏死率(P<0.05);九龙藤黄酮与程序性坏死抑制剂necrostatin-1合用有协同增效作用。结论九龙藤黄酮可抑制程序性坏死、抗心肌缺氧/复氧损伤,其机制可能与提高总抗氧化力,降低肿瘤坏死因子-α的含量,下调受体相互作用蛋白激酶3蛋白表达,降低细胞坏死率有关。
Objective To investigate the effects and mechanisms of Bauhinia championii flavones (BCF) on myocardial necrosis induced by hypoxia and reoxygenation. Methods The hypoxia / reoxygenation injury model of cardiomyocytes was established. Pretreatment with Jiulongng Flavonoids, morphological changes of myocardial cells were observed with inverted microscope. The total antioxidant capacity (T-AOC) was measured by UV spectrophotometry. Tumor necrosis factor (TNF-α). Western blot was used to detect the protein expression of receptor-mediated protein kinase 3 (RIPK3). The necrosis rate of cardiomyocytes was determined by Annex V-FITC / PI double staining. Results Compared with the model group, pretreatment with Jiulongtang flavonoids could reduce the injury of cardiomyocytes, increase the total antioxidant capacity, reduce the content of tumor necrosis factor-α, downregulate the protein kinase 3 protein expression and decrease the necrosis rate of cardiomyocytes (P <0.05). The synergistic effect of Jiulongng flavanone with necrostatin-1 was also observed. Conclusions Jiulong flavonoids can inhibit programmed necrosis and prevent myocardial hypoxia / reoxygenation injury. The mechanism may be related to the increase of total antioxidant capacity, the decrease of tumor necrosis factor-α, the down-regulation of receptor-protein kinase 3 protein expression, and the decrease of Cell necrosis rate.