朱洁　李泽庚　王保芹　童祥丽　彭青和

[摘要] 觀察不同低氧时段下，原代大鼠肺动脉平滑肌细胞（pulmonary arterial smooth muscle cells，PASMCs） ATP敏感性钾通道（ATPsensitive potassium channel，KATP通道）蛋白的表达，探讨芪白平肺胶囊含药血清（简称QBPF）调控PASMCs KATP通道蛋白表达与一氧化氮（nitric oxide，NO）的相关性。清洁级SD大鼠给予芪白平肺胶囊颗粒连续灌胃10 d，制备芪白平肺含药血清。采用组织块贴壁法，体外培养原代大鼠肺动脉平滑肌细胞；Western blot检测不同低氧时间下Kir6.1和SUR2B的表达，以及在一氧化氮合酶抑制剂——Nω硝基L精氨酸甲酯（NωnitroLarginine methyl ester，LNAME）和KATP通道抑制剂——格列本脲（glyburide，GLYB）分别干预下，QBPF对其表达的影响。低氧6 h后Kir6.1和SUR2B蛋白表达开始上调，在低氧24 h达到高峰，低氧48，72 h的蛋白表达出现不同程度下调。在低氧24 h条件下，QBPF能进一步上调KATP通道Kir6.1和SUR2B蛋白表达，且这种上调作用能分别被KATP通道阻断剂GLYB和NO特异性阻断剂LNAME所阻断，提示芪白平肺胶囊具有明确的KATP通道开放作用，其机制可能通过介导NO相关途径上调KATP通道蛋白表达，参与肺血管舒张作用，缓解COPD的发生发展。

[关键词] 肺动脉平滑肌细胞； ATP敏感性钾通道； 一氧化氮； 含药血清

Effect of Qibai Pingfei capsule medicated serum on protein expressions of

KATP channel in pulmonary arterial smooth muscle cells via nitric oxide

ZHU Jie1， LI Zegeng2，3*， WANG Baoqin4， TONG Xiangli4， PENG Qinghe1

（1. School of Integrated Traditional Chinese & Western Medicine， Anhui University of Chinese Medicine， Hefei 230038， China；

2. Anhui University of Chinese Medicine， Hefei 230038， China；

3. Institute of Medicine for Respiratory Disease， Anhui Academy of Chinese Medicine， Hefei 230038， China；

4. The First Affiliated Hospital， Anhui University of Chinese Medicine， Hefei 230031， China）

[Abstract] To investigate the ATPsensitive 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——NωnitroLarginine methyl ester（LNAME） 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 6hour hypoxia treament， peaked at the 24hour hypoxia treament， and decreased in both 48hour and 72hour hypoxia groups. Especially， QBPF could further upregulate the Kir6.1 and SUR2B protein expressions under 24hour hypoxia condition； however， such upregulation effect could be blocked by KATP channel inhibitor GLYB and NO specific inhibitor LNAME， indicating that QBPF played the role of opening KATP channel. The regulatory mechanism was probably associated with upregulating KATP channel protein expression via NO relative pathway， involving pulmonary vasodilation， and thus relieving the occurence and development of COPD.

[Key words] pulmonary vascular smooth muscle cells； ATPsensitive potassium channel； nitric oxide； medicated serum

慢性阻塞性肺疾病（chronic obstructive pulmonary disease，COPD）是一种发病率、致残率及病死率都很高的严重危害人民身体健康的慢性呼吸系统疾病[12]。肺动脉高压（pulmonary hypertesion，PH）是COPD的严重并发症之一，以肺血管阻力进行性增加和肺血管重构为主要特征[35]。益气化痰祛瘀方——芪白平肺胶囊（国家专利号ZL201010573274.1）用于治疗COPD，能有效缓解PH等并发症的进一步发展。前期研究表明，芪白平肺胶囊能有效上调一氧化氮（nitric oxide，NO）含量，改善低氧血症，舒张肺血管，但其具体作用途径有待进一步明确。……