银杏内酯B对阿霉素致心肌细胞凋亡的保护作用

Protective effects of ginkgolide B on adriamycin induced cardiomyocyte apoptosis

目的:探讨银杏内酯B对阿霉素损伤心肌细胞的保护作用及相关机制研究。方法:通过体外H9C2大鼠心肌细胞加入阿霉素5μmol/L构建损伤模型,将银杏内酯B 50μmol/L加入细胞中,共同培养,分为银杏内酯B组、银杏内酯B+阿霉素组、阿霉素组及空白对照组。通过FCM检测各组大鼠心肌细胞的凋亡率,通过Realtime PCR与Western blot技术检测蛋白激酶B(protein kinase B,AKT)、c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)、Caspase-3、Caspase-12 mRNA及蛋白表达水平。结果:阿霉素组H9C2细胞凋亡率为(46.23±4.98)%,银杏内酯B+阿霉素组凋亡率为(24.87±9.05)%,空白对照组凋亡率为(2.43±0.35)%,阿霉素组凋亡率较空白对照组显著升高,银杏内酯B+阿霉素组细胞凋亡率较阿霉素组显著降低,差异具有统计学意义(P<0.05)。Real-time PCR与Western blot检测结果显示,阿霉素能够上调H9C2细胞AKT、JNK、Caspase-3、Caspase-12的mRNA水平及蛋白水平表达(P<0.05),银杏内酯B+阿霉素组与阿霉素组相比,能够显著下调阿霉素损伤后H9C2细胞的AKT、JNK、Caspase-3、Caspase-12的mRNA水平及蛋白水平表达,差异均具有统计学意义(P<0.05)。结论:银杏内酯B可能通过抑制细胞凋亡的途径减轻阿霉素对心肌细胞的损伤,起保护心肌细胞的作用。

Objective： To investigate the protective effects of ginkgolide B on adriamycin induced myocardial injury and its related mechanism. Methods： The injury model was established by adding 5 μmol/L adriamycin into the H9C2 rat cardiomyocytes in vitro,and then 50 μmol/L ginkgolide B was added into the cells for co-culture. The cells were divided into the ginkgolide B group,ginkgolide B plus adriamycin group,adriamycin group and blank control group. The apoptosis rate of cardiomyocytes in each group was detected by flow cytometry. The mRNA and protein expression levels of AKT,JNK,Caspase-3 and Caspase-12 were detected by Real-time PCR and Western blot. Results： The apoptosis rate of adriamycin group was（ 46. 23 ± 4. 98） %,the apoptosis rate of ginkgolide B plus adriamycin group was（ 24. 87 ± 9. 05） %,and the apoptosis rate of blank control group was（ 2. 43 ± 0. 35） %. The apoptosis rate of H9C2 cells in adriamycin group was significantly higher than that of the normal control group,and the apoptosis rate of ginkgolide B plus adriamycin group was significantly lower than that of adriamycin group,with statistically significant differences（ P〈0.05）. The results of Real-time PCR and Western blot showed that adriamycin could increase the mRNA and protein expressions of AKT,JNK,Caspase-3 and Caspase-12 in H9C2 cells（ P〈0.05）. Compared with adriamycin group,ginkgolide B plus adriamycin group could significantly reduce the mRNA and protein expressions of AKT,JNK,Caspase-3 and Caspase-12 in H9C2 cells after adriamycin induced injury,with statistically significant differences（ P〈0.05）. Conclusion： Ginkgolide B may alleviate adriamycin induced cardiomyocytes injury by inhibiting apoptosis pathway,and play the protective effects on cardiomyocytes.