血根碱预处理对缺氧/复氧H9c2心肌细胞保护作用的实验研究

The Protective Effects on Sanguinarine on Hypoxia Reoxygenation H9c2 Cardiomyocytes

目的探讨血根碱对缺氧/复氧H9c2心肌细胞的保护作用,并探讨其机制。方法培养H9c2细胞,造成缺氧/复氧损伤模型,分为对照组、模型组、血根碱低浓度组及血根碱高浓度组,MTT法观察细胞活性,检测各组细胞上清液乳酸脱氢酶(LDH)及超氧化物歧化酶(SOD)、丙二醛(MDA)变化,流式细胞技术检测H9c2细胞内活性氧表达,Hoechst 33258染色观察凋亡细胞比例,蛋白免疫印迹法(Western Blot)分析Bcl-2及Bax蛋白表达的变化。结果低、高浓度血根碱干预后,H9c2细胞存活率逐渐提高,H9c2细胞凋亡率降低,细胞外培养基中LDH的释放量逐渐下降,Bcl-2蛋白、SOD表达水平升高,Bax蛋白、MDA表达水平降低,血根碱低浓度组、高浓度组与模型组比较差异均有统计学意义(P<0.05)。结论血根碱可以保护心肌细胞免受缺氧/复氧引起的损伤。

Objective To investigate the protective effect and mechanism of sanguinarine on hypoxia/reoxygenation H9c2 cells damage.Methods H9c2 cells were cultured and hypoxia/reoxygenation models were constructed,cells were divided into control group,model group,low concentration sanguinarine group,and high concentration sanguinarine group.Cell survival rate was detected by MTT colorimetry,the lactate dehydrogenase(LDH)in cell supernatant,superoxide dismutase(SOD),and malondialdehyde(MDA)in cells were measured,reactive oxygen species(ROS)expression inside the cells were detected by flow cytometry technology,apoptotic cells proportion were detected by Hoechst 33258 staining,the expression of Bcl-2 and Bax were tested by Western Blot.Results After treated with sanguinarine,H9c2 cells survival rate increased,the apoptosis rate of H9c2 cells decreased,the release of LDH in extracellular medium decreased gradually,the expression levels of Bcl-2 protein and SOD increased,the expression levels of Bax protein and MDA decreased.There were statistically significant differences between model group,low concentration sanguinarine group,and high concentration sanguinarine group(P<0.05).Conclusion Sanguinarine could inhibit hypoxia/reoxygenation-induced damage in H9c2 cells.