PI3K/Akt调控糖尿病冠状动脉平滑肌细胞BK通道的作用机制
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江苏省自然科学基金(BK20171151)


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

    Abstract:

    Objective:This study aims to investigate the effect of PI3K/Akt pathway on the regulation of large conductance Ca2+-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-α subunit 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 DCFH-DA. 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.

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焦国庆,李明秋,吴 莹,王如兴,胡亚玲,纪 丽,LU Tong. PI3K/Akt调控糖尿病冠状动脉平滑肌细胞BK通道的作用机制[J].南京医科大学学报(自然科学版),2022,42(2):166-170

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  • 收稿日期:2021-07-19
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  • 在线发布日期: 2022-03-01
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