容量激活性氯离子通道在低氧高二氧化碳处理的大鼠PASMCs的表达及其与MAPK通路的关系  

Expression of volume-activated chloride channel in rat PASMCs treated with hypoxia and hypercapnia and its relationship with MAPK pathway

作　　者：黎关龙[1,2] 黄林静[1] 何金波[1] 马迎春[1] 陈海娥[1] 应磊[1] 汪洋[1] 王万铁[1] 

机构地区：[1]温州医科大学基础医学院病理生理学教研室,浙江温州325035 [2]浙江省医学科学院卫生学研究所毒理学研究室,浙江杭州310013

出　　处：《中国病理生理杂志》2014年第1期25-29,共5页Chinese Journal of Pathophysiology

基　　金：浙江省自然科学基金资助项目(No.Y2080760);浙江省中医药重点学科建设计划项目(No.2012-XK-A28)

摘　　要：目的:探讨容量激活性氯离子通道(CLC3)在低氧高二氧化碳处理的大鼠肺动脉平滑肌细胞(PASMCs)中的表达变化及其与MAPK信号通路的关系。方法:酶消化法取雄性SD大鼠PASMCs进行原代培养,采用小鼠抗大鼠α-平滑肌肌动蛋白免疫荧光细胞化学法进行细胞鉴定;复制低氧高二氧化碳模型,采用免疫印迹法检测CLC3蛋白的表达;采用RT-PCR技术测定CLC3 mRNA水平的表达。结果:(1)与对照组比较,低氧高二氧化碳组PASMCs CLC3 mRNA和蛋白表达量均显著上调(均P<0.01);(2)与低氧高二氧化碳组比较,ERK抑制剂U0126+低氧高二氧化碳组PASMCs CLC3 mRNA和蛋白表达量均显著下调(均P<0.01);p38抑制剂SB203580+低氧高二氧化碳组PASMCs CLC3 mRNA和蛋白表达量均明显上调(均P<0.01);p38激活剂茴香霉素+低氧高二氧化碳组PASMCs CLC3 mRNA和蛋白表达水平均显著下降(P<0.05和P<0.01)。结论:低氧高二氧化碳可上调大鼠PASMCs CLC3 mRNA和蛋白的表达;ERK1/2通路介导了低氧高二氧化碳诱导的大鼠PASMCs CLC3表达,而p38 MAPK通路活化则下调低氧高二氧化碳诱导的CLC3 mRNA和蛋白表达。AIM： To investigate the expression of volume-activated chloride channel （CLC3） in rat pulmonary artery smooth muscle cells （PASMCs） treated with hypoxia and hypercapnia and its relationship with MAPK pathway. METHODS ： The method of enzyme digestion was used to isolate the PASMCs in male SD rat for cell primary culture. The cells were identified by cytochemical method with mouse anti-rat a-smooth muscle actin antibody. The rat model of hypoxia and hypercapnia was established. The protein expression of CLC3 was detected by Western blotting. The mRNA expression of CLC3 was determined by RT-PCR. RESULTS： Compared with control group, the mRNA and protein expression of CLC3 in PASMCs was significantly raised in hypoxia and hypercapnia group. Compared with hypoxic and hypercapnic group, the expression of CLC3 was significantly reduced in ERK inhibitor U0126 ＋ hypoxia and hypereap- nia group, and was up-regulated in p38 inhibitor SB203580 ＋ hypoxia and hypercapnia group, p38 activator anisomycin significantly decreased the expression of CLC3 at mRNA and protein levels in hypoxia and hypercapnia group. CONCLU- SION： The expression of CLC3 at mRNA and protein levels in PASMCs increases under hypoxia and hypercapnia condi-tions. The ERK1/2 pathway mediates CLC3 expression in PASMCs induced by hypoxia and hypercapnia. Activation of p38 MAPK pathway down-regulates the expression of CLC3 at mRNA and protein levels induced by hypoxia and hypercapnia.

关 键 词：低氧 高碳酸血症 容量激活性氯离子通道 p38 MAPK通路 ERKL 2通路 大鼠 

分 类 号：R363[医药卫生—病理学]