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目的:观察黄芪甲苷(AstragalosideⅣ,AsⅣ)后处理对缺氧复氧损伤(simulated ischemia reperfusion injury,SI/RI)的SD乳鼠心肌细胞是否具有保护作用。方法:将乳鼠原代心肌细胞平均分为五组,即空白对照组(Control)、缺氧复氧处理组(SI/RI)、黄芪甲苷预处理(5,10、20μM)+SI/RI组(AsIV+SI/RI)。各组细胞经处理后,四氮唑溴盐比色法(MTT)检测各组细胞存活率;TUNEL染色法测定各组细胞凋亡率;SOD测试盒检测培养液中超氧化物歧化酶(SOD)含量,总嘌呤氧化酶(XOD)测试盒检测丙二醛(MDA)含量。Western blot法检测各组细胞抗凋亡蛋白Bcl-2和促凋亡蛋白Caspase-3的表达。结果:与空白组相比,缺氧复氧损伤组细胞活力显著下降(P<0.05),凋亡率显著上升(P<0.05),其培养液中SOD水平显著降低(P<0.05),MDA水平显著升高。而不同浓度AsⅣ后处理组的心肌细胞存活率显著上升,凋亡率显著下降,培养液中SOD水平显著上升,MDA水平显著下降(P<0.05),且呈浓度呈依赖性。Western blot结果显示AsⅣ后处理组细胞中的Bcl-2表达明显上升,Caspase-3明显下降。结论:黄芪甲苷后处理对缺氧复氧诱导的乳鼠心肌细胞损伤具有显著的保护作用,能够显著上调抗凋亡蛋白Bcl-2的表达,下调促凋亡蛋白Caspase-3的表达。
Objective: To observe whether Astragaloside Ⅳ (As Ⅳ) can protect cardiomyocytes of neonatal SD rats after simulated ischemia reperfusion injury (SI / RI). Methods: Primary neonatal rat cardiomyocytes were divided into five groups randomly: control, hypoxia / reoxygenation group (SI / RI), Astragaloside pretreatment (5,10,20μM) + SI / RI group (AsIV + SI / RI). After treatment, the survival rate of each group was measured by tetrazolium bromide colorimetric method (MTT); the apoptosis rate of each group was determined by TUNEL staining; the SOD test kit was used to detect the activity of superoxide dismutase (SOD) Content, total purine oxidase (XOD) test box detection of malondialdehyde (MDA) content. Western blot was used to detect the expression of anti-apoptotic protein Bcl-2 and pro-apoptotic protein Caspase-3 in each group. Results: Compared with the blank group, the cell viability was significantly decreased (P <0.05) and the apoptosis rate was significantly increased (P <0.05). The SOD level in the culture medium was significantly lower (P <0.05) Significantly increased levels. However, the survival rate of cardiomyocytes in different concentrations of AsⅣ treated group increased significantly, and the apoptosis rate decreased significantly. The level of SOD in the culture medium increased significantly and the level of MDA decreased significantly (P <0.05) in a concentration - dependent manner. Western blot results showed that the expression of Bcl-2 in AsⅣ-treated cells was significantly increased and Caspase-3 significantly decreased. CONCLUSION: Astragaloside postconditioning has a significant protective effect on hypoxia-reoxygenation-induced cardiomyocyte injury in neonatal rats, which can significantly up-regulate the expression of anti-apoptotic protein Bcl-2 and down-regulate the expression of proapoptotic protein Caspase-3.