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背景:黄芩素对缺氧复氧损伤的心血管有保护作,但机制至今不清。目的:探讨中药黄芩素对心肌细胞缺氧/复氧损伤导致心肌细胞凋亡的保护作用机制。方法:体外培养大鼠乳鼠心肌细胞培养。实验分3组:正常对照组为正常培养的心肌细胞未做处理;缺氧/复氧组为应用缺氧/复氧方法诱导心肌细胞凋亡损伤;黄芩素预处理组为经黄芩素预处理30min后经缺氧/复氧诱导的心肌细胞。通过检测培养基上清液中乳酸脱氢酶活力检测细胞损伤程度及黄芩苷保护作用;应用原位末端标记细胞法标记细胞后,流式细胞检测心肌细胞凋亡率;应用免疫印迹方法检测心肌细胞凋亡蛋白Bax与抗凋亡蛋白Bcl-2的蛋白表达水平;应用Fura-2-AM负载心肌细胞,实时检测心肌细胞内Ca2+浓度变化。结果与结论:与正常对照组相比,缺氧/复氧组上清液乳酸脱氢酶活性、心肌细胞凋亡率、Bax蛋白含量、心肌细胞Ca2+浓度均增加(P<0.05),Bcl-2蛋白含量降低(P<0.05)。与缺氧/复氧组相比,黄芩素预处理组乳酸脱氢酶含量、心肌细胞凋亡率、Bax蛋白含量及心肌细胞Ca2+浓度均降低(P<0.05),Bcl-2蛋白含量增加(P<0.05)。证实黄芩素能抑制缺氧/复氧导致的心肌细胞凋亡,其作用机制可能与抗氧化与调节心肌细胞内钙离子浓度有关
BACKGROUND: Baicalein has protective effects on cardiovascular hypoxia-reoxygenation injury, but its mechanism remains unclear. Objective: To investigate the protective effect of baicalein on cardiomyocyte apoptosis induced by hypoxia / reoxygenation in cardiomyocytes. Methods: Rat neonatal rat cardiomyocytes were cultured in vitro. The experiment was divided into three groups: the normal control group was normal cultured cardiomyocytes without treatment; the hypoxia / reoxygenation group was induced by hypoxia / reoxygenation; the baicalein preconditioning group was pretreated with baicalein After 30min hypoxia / reoxygenation induced cardiomyocytes. The degree of cell injury and the protective effect of baicalin were detected by detecting the activity of lactate dehydrogenase in the supernatant of the culture medium. The apoptosis rate of cardiomyocytes was detected by flow cytometry after the cells were labeled with the in situ terminal labeling method. The protein expression levels of Bax and Bcl-2 were detected by real-time PCR. Fura-2-AM loaded cardiomyocytes were used to detect the intracellular Ca2 + concentration in real time. RESULTS AND CONCLUSION: Lactate dehydrogenase activity, myocardial cell apoptosis rate, Bax protein content and cardiomyocyte Ca2 + concentration in supernatant of hypoxia / reoxygenation group were increased (P <0.05), Bcl- 2 protein content decreased (P <0.05). Compared with the hypoxia / reoxygenation group, the content of Lactate dehydrogenase, the apoptosis rate of myocardial cells, the content of Bax protein and the content of Ca2 + in cardiomyocytes of baicalein preconditioning group were decreased (P <0.05), and the content of Bcl-2 protein increased P <0.05). Confirmed that baicalein can inhibit hypoxia / reoxygenation-induced cardiomyocyte apoptosis, its mechanism may be related to antioxidant and regulating the concentration of intracellular calcium