抑制蛋白激酶D1表达水平对高糖诱导心肌细胞凋亡的影响及作用机制  

The Effect and Its Mechanism of Inhibition of PKD1 Expression on Cardiomyocyte Apoptosis Induced by High Glucose

作　　者：杨佳伟[1] 胡沛[1] 谢进[1] 唐冰[1] YANG Jia-wei,HU Pei,XIE Jin,TANG Bing(Department of Cardiology,Jingzhou Central Hospital,Jingzhou 434020,China)

机构地区：[1]湖北省荆州市中心医院心内科,荆州434020

出　　处：《微循环学杂志》2018年第3期20-24,共5页Chinese Journal of Microcirculation

摘　　要：目的:探讨下调蛋白激酶D1(PKD1)对高糖环境下心肌细胞凋亡的影响及机制。方法:实验分5组:Control组(正常培养的H9c2细胞)、HG组(H9c2细胞高糖条件培养24h)、si-NC组(H9c2细胞转染siRNA control后高糖条件培养24h)、si-PKD1组(H9c2细胞转染PKD1siRNA后高糖条件培养24h)和si-PKD1+SB203580组[H9c2细胞转染PKD1siRNA 12h后加入p38丝裂原活化蛋白激酶(p38MAPK)信号通路抑制剂SB203580,再高糖条件培养24h],用qRT-PCR和Western blot方法检测各组细胞PKD1 mRNA和蛋白表达水平(si-PKD1+SB203580组除外);流式细胞术测定细胞凋亡情况,Western blot测定细胞促凋亡蛋白Bcl-2相关X蛋白(Bax)的表达水平,同时检测细胞中p38MAPK及其磷酸化p-p38MAPK水平。结果:HG组细胞PKD1mRNA和蛋白水平均明显高于Control组(P<0.01)。与HG组比较,si-PKD1组细胞中PKD1 mRNA和蛋白表达明显降低(P<0.01),细胞凋亡率降低,Bax蛋白表达水平下降,细胞中p-p38MAPK水平降低(P<0.01)。与si-PKD1组比较,siPKD1+SB203580组心肌细胞凋亡率进一步降低,细胞中Bax蛋白水平下降,p-p38MAPK、p-p38MAPK/p38MAPK水平降低(P<0.01)。结论:高糖诱导心肌细胞PKD1表达,下调PKD1可能通过抑制p38MAPK信号通路激活减少高糖诱导的心肌细胞凋亡。Objective：To study the effect and mechanism of down-regulation of PKD1 on cardiomyocyte apoptosis in high glucose environment.Method：The experiment was divided into 5 groups,Control group（normal cultured H9 c2 cells）,HG group（H9 c2 cells cultured in high glucose condition for 24 h）,si-NC group（H9 c2 cells transfected with siRNA control,then cultured in high glucose condition for 24 h）,si-PKD1 group（H9 c2 cells transfected with PKD1 siRNA,and then cultured in high glucose condition for 24 h）,and si-PKD1 ＋ SB203580 group（H9 c2 cells transfected PKD1 siRNA 12 h,p38 MAPK signal pathway inhibitor SB203580 was added to high glucose condition）.qRT-PCR and Western blot were used to detect the expression levels of PKD1 mRNA and protein in all the cells except for the si-PKD1＋SB203580.Flow cytometry was used to determine the apoptosis of cardiac myocytes,Western blot was used to determine the expression level of apoptotic protein Bax in cells,at the same time,the level of p38 MAPK phosphorylation in the cells was detected.Results：The levels of PKD1 mRNA and protein in HG group were significantly higher than those in Control group（P〈0.01）.The expression of PKD1 mRNA and protein in siPKD1 group was significantly lower than that in HG group（P〈0.01）,the rate of apoptosis was reduced,the expression level of Bax protein was decreased,the level of p-p38 MAPK in the cells was reduced,the difference was statistically significant（P〈0.01）.Compared with the si-PKD1 group,the apoptotic rate of myocardial cells in si-PKD1＋SB203580 group was further lower than that in si-PKD1 group,the level of Bax protein in the cells was decreased,the level of p-p38 MAPKand p-p38 MAPK/p38 MAPK were decreased（P〈0.01）.Conclusion：High glucose induces PKD1 expression in cardiac myocytes,down regulation of PKD1 can reduce the apoptosis of myocardial cells induced by high glucose by inhibiting the activation of p38 MAPK signaling pathway.

关 键 词：心肌细胞 PKD1 高糖 凋亡 

分 类 号：R542.2[医药卫生—心血管疾病]