当归红芪超滤物通过调控P53表达对H9C2细胞凋亡的干预效应研究

Study on the intervention effect of Radix Angelica Sinensis and Radix Hedysari ultrafiltration on H_(9)C_(2) cell apoptosis by regulating the expression of P53

目的:阐明P53高表达能否介导H_(9)C_(2)细胞凋亡及当归红芪超滤物(RAS-RH)能否通过调控P53表达干预H_(9)C_(2)细胞凋亡。方法:将H_(9)C_(2)细胞随机分为正常组,阴性组,模型组和RAS-RH低、中、高剂量组。采用CCK-8法检测各组细胞增殖率,EDU染色检测各组H_(9)C_(2)心肌细胞的凋亡,免疫荧光检测Bax、Bcl-2的表达,鬼笔环肽染色观察细胞骨架结构,Western Blot检测P53、Caspase-3、Caspase-8蛋白表达。结果:与阴性组比较,模型组H_(9)C_(2)细胞存活率显著降低(P<0.05);EDU标记的阳性细胞明显减少;Bax表达上调,Bcl-2表达下调;H_(9)C_(2)细胞微丝微管变短,排列紊乱;P53、Caspase-3、Caspase-8蛋白表达均显著上调(P<0.05)。与模型组比较,RAS-RH低、中、高剂量组H_(9)C_(2)细胞存活率显著增加(P<0.05);EDU标记的阳性细胞增多;Bax表达下调,Bcl-2表达上调;H_(9)C_(2)细胞微丝微管结构排列规则,扭曲、断裂减少;P53、Caspase-3、Caspase-8蛋白表达均显著降低(P<0.05)。结论:RAS-RH可通过下调P53表达抑制X线辐射诱导的H_(9)C_(2)细胞凋亡。

Objective:To clarify whether the high expression of P53 can mediate the apoptosis of H_(9)C_(2) cells and whether the ultrafiltration of Radix Angelica Sinensis and Radix Hedysari(RAS-RH)can interfere with the apoptosis of H_(9)C_(2) cells by regulating the expression of P53.Methods:H_(9)C_(2) cells were randomly divided into normal group,negative group,model group,and low,medium and high dose groups of RAS-RH.The proliferation rate of cells in each group was detected by CCK-8 method,the apoptosis of H_(9)C_(2) cardiomyocytes in each group was detected by EDU staining,the expressions of Bax and Bcl-2 were detected by immunofluorescence,and the cytoskeleton structure was observed by phalloidin staining.Western Blot was used to detect the expression of P53,Caspase-3 and Caspase-8 proteins.Results:Compared with the negative group,the survival rate of H_(9)C_(2) cells in the model group was significantly decreased(P<0.05),EDU-labeled positive cells were decreased,the expression of Bax was up-regulated and Bcl-2 down-regulated,H_(9)C_(2)cells were microfilament microtubules shortened and disordered,the protein expressions of P53,Caspase-3 and Caspase-8 were all up-regulated(P<0.05).Compared with the model group,the survival rate of H,C,cells in the RAS-RH low,medium and high dose groups was significantly increased(P<0.05);EDU marked positive cells were increased,Bax expression was down-regulated and Bcl-2 expression was up-regulated H_(9)C_(2) cells microfilaments and microtubules were arranged regularly,with fewer twists and breaks;the protein expressions of P53,Caspase-3 and Caspase-8 were all decreased(P<0.05).Conclusion:RAS-RH can significantly inhibit the apoptosis of H,C,cells induced by X-ray radiation by down-regulating the expressionof P53.