摘要
目的:探讨吴茱萸碱对心肌细胞缺血损伤的保护作用。方法:培养H9c2心肌细胞,缺氧24 h造成心肌细胞损伤模型,给予不同浓度(0.1,1,5,10μmol·L^(-1))吴茱萸预处理细胞12 h,CCK-8检测心肌细胞的活性,RT-PCR检测心肌细胞炎症因子的转录,Tunel染色检测心肌细胞凋亡,免疫印迹检测信号通路的改变。结果:4种浓度的吴茱萸碱在基础状态下并未影响心肌细胞活性(P>0.05);缺氧24 h后,心肌细胞活性显著降低,给予1,5,10μmol·L^(-1)的吴茱萸与对照组相比差异有统计学意义(P<0.05),且其效果呈现剂量依耐性;细胞促炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素-1(IL-1)和白细胞介素-6(IL-6)的转录明显增加,心肌细胞凋亡增多,吴茱萸碱可以增加缺氧损伤的细胞活性,减少细胞炎症因子的转录,减少细胞凋亡数量,给予1μmol·L^(-1)和10μmol·L^(-1)的吴茱萸与对照组相比差异有统计学意义(P<0.05)。免疫印迹结果显示,吴茱萸碱通过增加蛋白激酶B(AKT)和AMP依赖蛋白激酶α(AMPKα)的活性,抑制核因子-κB(NF-κB)的活性发挥其心肌保护作用,给予1μmol·L^(-1)和10μmol·L^(-1)的吴茱萸与对照组相比差异有统计学意义(P<0.05)。结论:吴茱萸碱能保护心肌细胞缺血损伤,可能成为新的抗心肌缺血药物。
Objective：To investigate the protective effect of evodiamine on cardiomyocytes hypoxia injury.Methods：H9c2 myocardial cells were exposed to hypoxia for 24 h to induce myocardial cell injury model.The cells were pretreated with different concentrations of evodiamine for 12 h.The cardiomyoctes viability was detected by CCK-8 assay.The transcription of inflammatory cytokines was detected by RT-PCR.The cardiomyocyte apoptosis was detected with Tunel staining.The alteration of signal pathway was detected by immunoblotting.Results：Four concentrations of evodiamine did not affect the activity of cardiomyocytes in the basal condition（P0.05）.After 24-hour hypoxia,the viability of cardiomyocytes decreased significantly when compared with that in the control group with significant difference（P0.05）among 1,5 and 10μmol·L^（-1）evodia in a dose-dependent manner.The transcription of pro-inflammatory cytokines（TNF-α,IL-1 and IL-6）significantly increased and the cells apoptosis increased.Evodiamine pretreatment increased the cell viability after hypoxia injury,reduced the transcription of inflammatory cytokines and reduced the number of apoptotic cells when compared with that in the control group with significant differences（P0.05）between 1μmol·L^（-1）and 10μmol·L^（-1）evodia.The results of western blot showed that evodiamine activated AMPKαand AKT,inhibited the activity of NF-kappa B,and compared with the control group,there were significant differences（P0.05）between 1μmol·L^（-1） and 10μmol·L^（-1） of evodia.Conclusion：Evodiamine can protect cardiomyocytes from hypoxia injury and may become a new anti-myocardial ischemia drug.
出处
《中国药师》
CAS
2018年第2期193-197,共5页
China Pharmacist
基金
国家自然科学基金项目(编号:81700353)
