PI3K/Akt调控糖尿病冠状动脉平滑肌细胞BK通道的作用机制  

PI3K/Akt pathway mediates large conductance calcium-activated potassium in diabetic coronary smooth muscle cells

作　　者：焦国庆 李明秋 吴莹 王如兴 胡亚玲 纪丽 LU Tong JIAO Guoqing;LI Mingqiu;WU Ying;WANG Ruxing;HU Yaling;JI Li;LU Tong(Wuxi Institute of Translational Medicine;Department of Cardiology,Wuxi People’s Hospital Affiliated to Nanjing Medical University,Wuxi 214023,China;Division of Cardiovascular Diseases,Mayo Clinic,Rochester 55905,USA)

机构地区：[1]无锡市转化医学研究所 [2]南京医科大学附属无锡人民医院心脏外科,江苏无锡214023 [3]Mayo Clinic细胞电生理研究室,美国明尼苏达州罗切斯特55905

出　　处：《南京医科大学学报（自然科学版）》2022年第2期166-170,199,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省自然科学基金(BK20171151)。

摘　　要：目的:探讨PI3K/Akt对糖尿病冠状动脉平滑肌细胞BK通道的调节作用。方法:采用正常糖浓度(5 mmon/L)、高糖浓度(15 mmon/L)、活性氧(reactive oxygen species,ROS)抑制剂GKT137831、PI3K抑制剂Wortmannin作用于人冠状动脉平滑肌细胞(human coronary artery smooth muscle cell,HCASMC)。以GKT137831喂饲链脲霉素(streptozocin,STZ)诱导的糖尿病大鼠,免疫印迹测定HCASMC、大鼠冠状动脉PI3K超家族DNA-Pkcs、Akt、p-Akt(S473)及BK-α亚基、BK-β1亚基的表达,2’,7’-二氯二氢荧光素二乙酸酯测定HCASMC及大鼠冠状动脉ROS表达。结果:高糖浓度下HCASMC及糖尿病大鼠冠状动脉ROS、DNA-Pkcs、Akt、p-Akt(S473)表达量增高,BK-β1表达量降低(P<0.05)。GKT137831作用后的HCASMC及糖尿病大鼠冠状动脉中,ROS、DNA-Pkcs、Akt、p-Akt(S473)表达量降低,BK-β1表达量明显增高(P<0.05)。Wortmannin作用后的HCASMC p-Akt(S473)表达量降低,BK-β1表达量明显增加(P<0.05)。结论:PI3K/Akt通路参与糖尿病冠状动脉平滑肌细胞BK通道的调控。Objective:This study aims to investigate the effect of PI3K/Akt pathway on the regulation of large conductance Ca;-activated K+channel(BK channel)in diabetic coronary smooth muscle cells.Methods:Human coronary artery smooth muscle cells(HCASMC)were incubated with different concentrations of glucose(5 mmol/Land 15 mmol/L),reactive oxygen species(ROS)inhibitor GKT137831 and PI3K inhibitor wortmannin.Streptozotocin-induced diabetic rats were established successfully by intraperitoneal injection.Expression of DNA-Pkcs,Akt,p-Akt(S473),BK-αsubunit and BK-β1 subunits were determined by Western blot.ROS generation in HCASMC was measured in terms of fluorescence using the cell-permeable fluorogenic probe DCFHDA.Results:Compared with that of HCASMC incubated in 5 mmon/L glucose concentration,the expression of DNA-Pkcs,Akt,p-Akt(S473)increased significantly and BK-β1 subunit decreased significantly(P<0.05).Compared with the control group,the expression of p-Akt(s473)decreased significantly(P<0.05)and BK-β1 increased significantly(P<0.05)after pretreated with Wortmannin.Conclusion:PI3K/Akt channel is involved in the regulation of BK channel expression in diabetic coronary smooth muscle cells.

关 键 词：糖尿病 大电导钙激活钾通道β1亚基 冠状血管 

分 类 号：R329.26[医药卫生—人体解剖和组织胚胎学]