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目的:研究葛根素对表柔比星致H9c2心肌细胞凋亡的影响。方法:取H9c2心肌细胞分为空白对照组、模型组、阳性对照组(10 μmol/L 右丙亚胺)和葛根素低、中、高浓度组(10、50、100 μmol/L),每组5个复孔。除空白对照组不作任何处理外,其他各组细胞加入表柔比星(1.0 μmol/L)及相应药物,作用24 h后,采用CCK-8法检测细胞存活率,Hoechst 33258染色观察细胞的凋亡形态,流式细胞术检测细胞凋亡率,Western blot 法检测凋亡相关信号蛋白Bax、Bcl-2、Cleaved-caspase-3的表达,比色法检测超氧化物歧化酶(SOD)活性及丙二醛(MDA)的含量。结果:与空白对照组比较,模型组细胞核出现明显的凋亡小体,细胞存活率、Bcl-2表达、SOD活性均降低,凋亡率、Bax和Cleaved-caspase-3表达、Bax/Bcl-2比例、MDA含量均增加;与模型组比较,阳性对照组和葛根素中、高浓度组细胞核形态完整,细胞存活率、Bcl-2表达、SOD活性均增加,凋亡率、Bax和Cleaved-caspase-3表达、Bax/Bcl-2比例、MDA含量均降低,差异均有统计学意义(P<0.05)。其余指标差异无统计学意义(P>0.05)。结论:葛根素可能通过抑制细胞凋亡信号通路和减轻氧化损伤作用来降低表柔比星的心肌毒性。
OBJECTIVE: To study the effects of puerarin (Pue) on the apoptosis of H9c2 myocardial cell induced by epirubicin(Epi). METHODS: The H9c2 myocardial cells were divided into blank control group, model group, positive control group (10 μmol/L dexrazoxane) and Pue low-dose, medium-dose and high-dose groups (10, 50, 100 μmol/L), with 5 wells in each group. Except blank control group didn’t received any treatment, other groups were treated with Epi (1.0 μmol/L) and relevant medicine. 24 h later, survival rate of myocardial cell was measured by CCK-8 kit; apoptotic morphology was observed by Hoechst 33258 fluorescent staining; apoptotic rate of myocardial cell was determined by flow cytometry; the protein expression of Bax, Bcl-2 and Cleaved-caspase-3 were assayed by Western blot; SOD activity and MDA content were assayed by colorimetric kit. RESULTS: Compared with blank control group, apoptotic body was obviously found in cell nucleus of model group; survival rate, the expression of Bcl-2 and SOD activity were decreased; while the apoptotic rate, the expression of Bax and Cleaved-caspase-3, Bax/Bcl-2 ratio and MDA content were increased. Compared with model group, the morphology of cell nucleus was complete in positive control group and Pue medium-dose and high-dose groups; survival rate, the expression of Bcl-2 and SOD activity were increased; while apoptotic rate, the expression of Bax and Cleaved-caspase-3, Bax/Bcl-2 ratio and MDA content were decreased, with statistical significance (P<0.05). There was no statistical significance in other indexes (P>0.05). CONCLUSIONS: Pue could mitigate cardiac toxicity of Epi via inhibiting apoptotic signaling pathway and depressing oxidative damage.
葛根素表柔比星H9c2心肌细胞心肌毒性
PuerarinEpirubicinH9c2 myocardial cellCardiac toxicity
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