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低氧高二氧化碳对大鼠肺动脉平滑肌细胞KATP通道表达的影响及其与p38 MAPK通路的关系

马迎春, 黄林静, 郑梦晓, 王园园, 应磊, 王万铁

温州医科大学基础医学院病理生理学教研室；缺血-再灌注损伤研究所，温州 325035；浙江省立同德医院病理科，杭州 310012

摘要

本文旨在探究ATP敏感性钾通道(KATP)在大鼠低氧高二氧化碳性肺动脉平滑肌细胞(pulmonary artery smooth muscle cells, PASMCs)中的表达及其与p38 MAPK信号通路的关系。体外培养SD大鼠PASMCs，复制低氧高二氧化碳模型，采用RT-PCR技术及免疫印迹法检测KATP亚基磺酰脲类受体2B (sulfonylurea receptor 2B, SUR2B)和内向整流钾通道6.1 (Kir6.1) mRNA及蛋白的表达水平。结果显示：(1)与常氧组(N组)、低氧高二氧化碳组(H组)、低氧高二氧化碳+溶剂DMSO对照组(HD组)、低氧高二氧化碳+p38 MAPK通路抑制剂SB203580组(HS组)相比，低氧高二氧化碳+p38 MAPK通路激动剂茴香霉素(Anisomycin)组(HA组) Kir6.1 mRNA与蛋白表达均明显降低(P < 0.01)，N组、H组、HD组、HS组间Kir6.1 mRNA与蛋白表达差异不显著(P > 0.05)；(2)与N组相比，H组、HD组、HS组、HA组SUR2B mRNA与蛋白表达均明显上升(P < 0.05)，H组、HD组、HS组、HA组间SUR2B mRNA与蛋白表达差异不显著(P > 0.05)。以上结果提示：(1)低氧高二氧化碳、SB203580均没有引起Kir6.1 mRNA与蛋白表达变化，而茴香霉素下调Kir6.1 mRNA与蛋白表达，Kir6.1可能受其它类型的MAPK通路的调节；(2)低氧高二氧化碳明显上调SUR2B mRNA与蛋白表达，而SB203580、茴香霉素不影响低氧高二氧化碳所引起的SUR2B表达上调，低氧高二氧化碳引起的SUR2B表达的升高可能是非p38 MAPK通路依赖性的。

关键词： 低氧高二氧化碳; ATP敏感性钾通道; 内向整流钾通道6.1; 磺酰脲类受体2B; p38丝裂原活化蛋白激酶; 大鼠

分类号：R363

[Expression of KATP in pulmonary artery smooth muscle cells under hypoxiahypercapnia condition and the relationship with p38 MAPK pathway.] [Article in Chinese]

MA Ying-Chun； HUANG Lin-Jing； ZHENG Meng-Xiao； WANG Yuan-Yuan； YING Lei, WANG Wan-Tie

Department of Pathophysiology； Institute of Ischemia/Reperfusion Injury, Wenzhou Medical University, Wenzhou 325035, China

Abstract

The aim of the present study is to investigate the expressions of ATP-sensitive K⁺ channels (K_ATP) in pulmonary artery smooth muscle cells (PASMCs) and the relationship with p38 MAPK signal pathway in rats. Male SD rat PASMCs were cultured in vitro, and a model of hypoxia and hypercapnia was reconstructed. PASMCs were divided to normal (N), hypoxia-hypercapnia (H), hypoxia-hypercapnia+DMSO incubation (HD), hypoxia-hypercapnia+SB203580 (inhibitor of p38 MAPK pathway) incubation (HS) and hypoxia-hypercapnia+Anisomycin (agonist of p38 MAPK pathway) incubation (HA) groups. Western blot was used to detect the protein expression of SUR2B and Kir6.1; semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of SUR2B and Kir6.1. The results demonstrated that: (1) Compared with N, H, HD and HS groups, the expressions of Kir6.1 mRNA and protein in PASMCs of HA group were decreased significantly (P < 0.01), but there were no differences among N, H, HD and HS groups (P > 0.05); (2) Compared with N group, the expressions of SUR2B mRNA and protein in H, HD, HS and HA groups were increased significantly (P < 0.05), but there were no differences among H, HD, HS and HA groups (P > 0.05). The results imply that: (1) Hypoxia-hypercapnia, SB203580 didn't change the expressions of Kir6.1 mRNA and protein in PASMCs, but Anisomycin decreased the expressions of Kir6.1 mRNA and protein, so Kir6.1 may be regulated by the other subfamily of MAPK pathway; (2) Hypoxia-hypercapnia raised SUR2B mRNA and protein expressions in PASMCs, but SB203580 and Anisomycin did not affect the changes, so the increasing of SUR2B mRNA and protein induced by hypoxia-hypercapnia may be not depend on p38 MAPK pathway.

Key words: hypoxia hypercapnia; KATP; Kir6.1; SUR2B; p38 MAPK; rat

收稿日期：2013-10-30　　录用日期：2014-02-10

通讯作者：王万铁　　E-mail: wwt@wzmc.edu.cn

引用本文：

马迎春, 黄林静, 郑梦晓, 王园园, 应磊, 王万铁. 低氧高二氧化碳对大鼠肺动脉平滑肌细胞KATP通道表达的影响及其与p38 MAPK通路的关系[J]. 生理学报 2014; 66 (3): 283-288.

MA Ying-Chun； HUANG Lin-Jing； ZHENG Meng-Xiao； WANG Yuan-Yuan； YING Lei, WANG Wan-Tie. [Expression of KATP in pulmonary artery smooth muscle cells under hypoxiahypercapnia condition and the relationship with p38 MAPK pathway.] [Article in Chinese]. Acta Physiol Sin 2014; 66 (3): 283-288 (in Chinese with English abstract).