Zhu Jie, Li Ze-Geng, Wang Bao-Qin, Tong Xiang-Li, Peng Qing-He
School of Integrated Traditional Chinese & Western Medicine, Anhui University of Chinese Medicine, Hefei 230038, China.
Anhui University of Chinese Medicine, Hefei 230038, China.
Zhongguo Zhong Yao Za Zhi. 2017 Apr;42(7):1376-1381. doi: 10.19540/j.cnki.cjcmm.2017.0059.
To investigate the ATP-sensitive potassium channel (KATP channel) protein expressions during different periods under hypoxia condition and explore the effect of Qibai Pingfei capsule medicated serum (hereinafter referred to as QBPF) on the correlation between the protein expressions of KATP channel and nitric oxide in rat pulmonary arterial smooth muscle cells(PASMCs). Qibai Pingfei capsules were given to SD rats via continuous gavage for 10 days to obtain QBPF. Primary rats PASMCs were cultured by the direct adherent culture method. Western blot was applied to detect the protein expression levels of KATP channel (Kir6.1 and SUR2B) in PASMCs. Then the noncompetitive inhibitor of NO synthase--Nω-nitro-L-arginine methyl ester(L-NAME) and KATP channel inhibitor--glyburide(GLYB) were applied respectively to evaluate the effect of QBPF on the protein expressions of KATP channel. The protein expressions of Kir6.1 and SUR2B were increased after 6-hour hypoxia treament, peaked at the 24-hour hypoxia treament, and decreased in both 48-hour and 72-hour hypoxia groups. Especially, QBPF could further up-regulate the Kir6.1 and SUR2B protein expressions under 24-hour hypoxia condition; however, such up-regulation effect could be blocked by KATP channel inhibitor GLYB and NO specific inhibitor L-NAME, indicating that QBPF played the role of opening KATP channel. The regulatory mechanism was probably associated with up-regulating KATP channel protein expression via NO relative pathway, involving pulmonary vasodilation, and thus relieving the occurence and development of COPD.
研究缺氧条件下不同时段ATP敏感性钾通道（KATP通道）蛋白的表达情况，探讨芪百平喘胶囊含药血清（以下简称QBPF）对大鼠肺动脉平滑肌细胞（PASMCs）中KATP通道蛋白表达与一氧化氮之间相关性的影响。通过连续灌胃给予SD大鼠芪百平喘胶囊10天以获取QBPF。采用直接贴壁培养法培养原代大鼠PASMCs。应用蛋白质免疫印迹法检测PASMCs中KATP通道（Kir6.1和SUR2B）的蛋白表达水平。然后分别应用一氧化氮合酶的非竞争性抑制剂——Nω-硝基-L-精氨酸甲酯（L-NAME）和KATP通道抑制剂——格列本脲（GLYB）来评估QBPF对KATP通道蛋白表达的影响。缺氧处理6小时后Kir6.1和SUR2B的蛋白表达增加，在缺氧处理24小时时达到峰值，而在缺氧48小时和72小时组中均下降。特别地，QBPF可在24小时缺氧条件下进一步上调Kir6.1和SUR2B的蛋白表达；然而，这种上调作用可被KATP通道抑制剂GLYB和一氧化氮特异性抑制剂L-NAME阻断，表明QBPF发挥了开放KATP通道的作用。其调节机制可能与通过一氧化氮相关途径上调KATP通道蛋白表达有关，涉及肺血管舒张，从而缓解慢性阻塞性肺疾病的发生和发展。
