吡那地尔对人肺动脉平滑肌细胞KATP通道mRNA表达的影响
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江苏省自然科学基金资助项目(BK2006246)


Effects of Pinacidil on mRNA expression of KATP channels of human pulmonary artery smooth muscle cells
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    目的:研究经典KATP开放剂吡那地尔对原代培养人肺动脉平滑肌细胞KATP通道mRNA表达的影响,以探讨KATP通道在肺动脉高压发生发展中的分子生物学机制-方法:分离人3~4级肺小动脉,原代培养人肺动脉平滑肌细胞-分成ET-1组-ET-1+吡那地尔组及ET-1+吡那地尔+格列本脲组-以Trizol法抽提总RNA,逆转录为cDNA,采用Real-time PCR方法检测各组细胞KATP通道SUR2B和Kir亚单位 mRNA表达量-结果:ET-1组KATP通道SUR2B mRNA表达量较对照组明显减少,为对照组的0.09 ± 0.01倍(P < 0.05,n = 3)-加用吡那地尔可拮抗ET-1作用,提高SUR2B mRNA表达量,为ET-1组的10.94 ± 1.13倍,为对照组的0.97 ± 0.03倍(P > 0.05,n = 3)-格列本脲可拮抗吡那地尔作用,减少SUR2B mRNA表达量,为ET-1+吡那地尔组的0.10 ± 0.01倍,为对照组的0.07 ± 0.02倍(P < 0.05,n = 3)-各组细胞KATP通道Kir6.1 mRNA表达量无统计学差异-结论:KATP开放剂吡那地尔可增加原代培养人肺动脉平滑肌细胞KATP通道SUR2B mRNA表达量,这可能是其调节肺动脉平滑肌细胞KATP通道表达和功能的机制-

    Abstract:

    Objective:To study the effect of Pinacidil on mRNA expression of ATP-sensitive K+(KATP) channels of primary cultured human pulmonary artery smooth muscle cells. Methods:Intrapulmonary arteries(3rd~4th division) were opened longitudinally. After removing the adventitia and epithelium by a blade,they were cut into pieces of 1~3 mm2. The tissue and smooth muscle cells were cultured with DMEM. Cells were incubated with test substances for 24 h by deviding into following groups:Control,ET-1 10 nmol/L,ET-1+Pinacidil 10 μmol/L,ET-1+Pinacidil 10 μmol/L+Glyburide 10 μmol/L. Total RNA was extracted by Trizol and reverse transcribed into cDNA. Real-time PCR was explored to measure the level of mRNA expression of KATP channels. Results:ET-1 suppressed expression of SUR2B mRNA of KATP channels for 0.09 ± 0.01-fold(P < 0.05,n = 3),compared with control group. Expressions of SUR2B mRNA in the group of ET-1+Pinacidil 10 μmol/L were 0.97 ± 0.03-fold(P > 0.05,n = 3) compared with control group. ET-1+Pinacidil 10 μmol/L+Glyburide 10 μmol/L were 0.07 ± 0.02-fold(P < 0.05,n = 3),compared with control group. There were no significant differences in the expression of Kir6.1 mRNA among all groups. Conclusion:Pinacidil increased the expression KATP channel SUR2B mRNA of primary cultured human pulmonary artery smooth muscle cells. This might be the mechanism in regulating mRNA expression of KATP channels in pulmonary arteries.

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李庆玲,周燕娟,解卫平,王 虹,朱煜明.吡那地尔对人肺动脉平滑肌细胞KATP通道mRNA表达的影响[J].南京医科大学学报(自然科学版),2008,28(5):585-588

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  • 收稿日期:2007-11-20
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