柚皮素对缺氧损伤大鼠心肌细胞的保护作用  

Protective effect of naringenin on hypoxia-injured myocardial cells in rats

作　　者：李磊 胡立群[1] 邹丽绢 赵鑫扬 彭晟 吴刚 LI Lei;HU Li-qun;ZOU Li-juan;ZHAO Xin-yang;PENG Sheng;WU Gang(Department of Cardiology,Wuhan Fourth Hospital(Puai Hospital,Tongji Medical College,Huazhong University of Science and Technology),Wuhan 430033,Hubei,China)

机构地区：[1]武汉市第四医院(华中科技大学同济医学院附属普爱医院)心内科,武汉430033

出　　处：《第二军医大学学报》2021年第6期657-663,共7页Academic Journal of Second Military Medical University

基　　金：2018年武汉市卫生健康委员会项目(WZ18Q10,WX18Q02)。

摘　　要：目的探究柚皮素对缺氧损伤的大鼠心肌细胞是否有保护作用。方法用大鼠心肌细胞系H9c2细胞制备心肌细胞缺氧损伤模型,并用乳酸脱氢酶(LDH)细胞毒性检测试剂盒进行模型鉴定。实验分为5组:对照组,模型组,以及柚皮素低、中、高剂量组(20、40、80μmol/L)。用CCK-8检测细胞增殖抑制率,间接免疫荧光法检测细胞肥大状况,流式细胞术检测细胞凋亡水平,蛋白质印迹法检测凋亡相关蛋白Bcl-2、Bcl-2关联X蛋白(Bax)、caspase 3的表达水平,qRT-PCR检测血管内皮细胞生长因子(VEGF)、NK2型同源异形框基因5(Nkx2.5)、α-平滑肌肌动蛋白(α-SMA)的mRNA表达水平。结果模型组细胞培养上清中LDH活性高于对照组(P<0.01),表明缺氧损伤心肌细胞建模成功。柚皮素各剂量组细胞增殖抑制率均低于模型组(P<0.05、P<0.01)。与模型组比较,柚皮素各剂量组H9c2心肌细胞肥大状况有所缓解(P<0.05、P<0.01)。柚皮素中、高剂量组凋亡细胞比例均较模型组降低(P均<0.01)。与模型组比较,柚皮素各剂量组凋亡蛋白caspase 3、Bax表达均降低,凋亡抑制蛋白Bcl-2表达均升高(P<0.05、P<0.01)。与模型组比较,柚皮素高剂量组Nkx2.5 mRNA表达水平升高(P<0.01),柚皮素中、高剂量组VEGF mRNA表达水平均升高(P<0.05、P<0.01),柚皮素各剂量组α-SMA mRNA表达水平均降低(P均<0.01)。结论柚皮素对缺氧损伤的大鼠心肌细胞有保护作用。Objective To investigate the protective effect of naringenin on hypoxia-injured myocardial cells.Methods A hypoxia-injured myocardial cell model was established with rat myocardial cell line H9c2 cells and identified by lactate dehydrogenase(LDH)cytotoxicity test kit.The experiment was divided into 5 groups:control group,model group,and naringenin low-,medium-and high-dose groups(20,40 and 80μmol/L).Cell proliferation inhibition rate was detected by cell counting kit 8;cell hypertrophy was detected by indirect immunofluorescence;cell apoptosis was detected by flow cytometry;the protein expression of apoptosis-related proteins B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax)and caspase 3 was detected by Western blotting;and the mRNA expression of vascular endothelial growth factor(VEGF),NK2 homeobox 5(Nkx2.5)and alpha-smooth muscle actin(α-SMA)in H9c2 cells was detected by qRT-PCR.Results Compared with the control group,the activity of LDH in cell supernatant of the model group was significantly increased(P<0.01),indicating that the hypoxia-injured myocardial cell model was successfully constructed.Compared with the model group,the inhibition rate of cell proliferation was significantly decreased in each dose group of naringenin(P<0.05,P<0.01);the hypertrophy of H9c2 cells in each dose group of naringenin was significantly alleviated(P<0.05,P<0.01);and the proportion of apoptotic cells was significantly decreased in the medium-and high-dose naringenin groups(both P<0.01).Compared with the model group,the expression levels of apoptotic proteins caspase 3 and Bax in each dose group of naringenin were significantly decreased,while the expression levels of apoptosis inhibition-related protein Bcl-2 were significantly increased(P<0.05,P<0.01).Compared with the model group,the mRNA expression level of Nkx2.5 in high-dose naringenin group was significantly increased(P<0.01),the mRNA expression levels of VEGF were significantly increased in medium-and high-dose naringenin groups(P<0.05,P<0.01),and the mRNA expre

关 键 词：柚皮素 心肌细胞 细胞增殖 细胞凋亡 凋亡调节蛋白质类 

分 类 号：R542.22[医药卫生—心血管疾病]