钙激活性氯离子通道在大鼠低氧高二氧化碳性PASMCs中的表达及与MAPK通路的关系  

The Expression of Calcium-activated Chloride Channel in PASMCs' under Hypoxic Hypercapnia State and the Relationship with MAPK Pathways

作　　者：黄林静[1] 黎关龙[2] 何金波[1] 马迎春[1] 陈丹[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,温州325035 [2]浙江省医学科学院卫生学研究所毒理学研究室,杭州310013

出　　处：《中国细胞生物学学报》2013年第9期1321-1327,共7页Chinese Journal of Cell Biology

基　　金：浙江省自然科学基金(批准号:Y2080760);浙江省中医药重点学科建设计划项目(批准号:2012-XK-A28)资助的课题~~

摘　　要：采用酶消化法取雄性SD大鼠肺动脉平滑肌细胞(pulmonary artery smooth muscle cells,PASMCs)进行原代培养,采用小鼠抗大鼠SMα-actin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钙激活性氯离子通道(calcium activated chloride channel,CLCA2)蛋白的表达;采用半定量逆转录–聚合酶链反应(RT-PCR)技术测定CLCA2 mRNA水平的表达。结果显示:与正常组比较,低氧高二氧化碳组PASMCs中CLCA2 mRNA和蛋白表达水平均明显上调(P<0.01);与低氧高二氧化碳组比较,U0126组和SB203580组PASMCs中CLCA2 mRNA和蛋白表达水平均明显上调(P<0.01);茴香霉素(Anisomycin)组PASMCs中CLCA2 mRNA和蛋白表达水平均显著下调(P<0.05和P<0.01)。研究表明,低氧高二氧化碳可上调大鼠PASMC中CLCA2 mRNA和蛋白的表达;ERK1/2通路抑制剂(U0126)、P38MAPK通路抑制剂(SB203580)均可上调PASMCs中CLCA2 mRNA和蛋白的表达;MAPK通路激活剂(Anisomycin)能下调PASMCs中CLCA2 mRNA和蛋白的表达。We adopt an enzyme digestion to take the male SD rat pulmonary artery smooth muscle cells （PASMCs） for cell primary culturing, taking immunofluorescence cytochemical method with mouse anti-rat the SM α-actin for cell identification. Then we set up a model of hypoxia and hypercapnia. Western blot is used to detect the protein expression of calcium-activated chloride channel （CLCA2）; We Use semi-quantitative reverse transcription- polymerase chain reaction to detecte the mRNA expression of CLCA2. The results demonstrated that compared with normal group, the expressions of CLCA2 mRNA and protein in PASMCs were significantly raised in hypoxiahypercapnia group （P〈0.01）; Contrast to the hypoxic hypercapnic group, CLCA2 mRNA and protein expressions in PASMCs were significantly raised in group U and group SB （P〈0.01）; the mRNA and protein expression in PASMCs were significantly decreased in group A （P〈0.05 and P〈0.01）. The expression of CLCA2 in PASMCs could be raised under hypoxia and hypercapnia conditions; The expression of CLCA2 in PASMCs could be raised by both U0126 and SB203580; The expression of CLCA2 in PASMCs could be decreased by Anisomycin.

关 键 词：低氧高二氧化碳 钙激活性氯离子通道 P38MAPK ERK1/2 大鼠 

分 类 号：R341[医药卫生—基础医学]