气道平滑肌细胞通过TGF-β1/Smad3信号通路调节IL-33的表达参与哮喘  

Airway smooth muscle cells regulate IL-33 expression through TGF-β1/Smad3 signaling pathway to participate in asthma

作　　者：蔡秋景 张倩[1] 何学佳 孙文丽 郭爱丽[1] 张楠[1] 朱薇薇[1] CAI Qiujing;ZHANG Qian;HE Xuejia;SUN Wenli;GUO Aili;ZHANG Nan;ZHU Weiwei(Department of Pediatrics,Jinan Central Hospital Affiliated to Shandong University,Jinan 250013,Shandong,China)

机构地区：[1]山东大学附属济南市中心医院儿科,山东济南250013

出　　处：《山东大学学报（医学版）》2020年第4期78-83,共6页Journal of Shandong University:Health Sciences

摘　　要：目的探究小鼠气道平滑肌细胞表达与分泌白细胞介素(IL)-33参与哮喘的信号机制。方法观察不同浓度[0 ng/mL(空白)、1 ng/mL、10 ng/mL、100 ng/mL]TGF-β1组对小鼠气道平滑肌细胞分泌表达IL-33的影响,ELISA法检测各组细胞培养上清液中IL-33的浓度,Western blotting检测各组细胞IL-33的蛋白表达量;观察加入TGF-β1/Smad3信号通路阻断剂(SIS3)后对该过程的抑制作用,分为空白组、预处理TGF-β1组、未预处理SIS3组和预处理SIS3组,ELISA法检测各组细胞培养上清中IL-33浓度,Western blotting检测各组Smad3、pSmad3及IL-33蛋白表达量。结果ELISA结果表明,空白组、1 ng/mL TGF-β1组、10 ng/mL TGF-β1组和100 ng/mL TGF-β1组的细胞上清IL-33浓度的总体差异有统计学意义(F=106.4,P<0.05);与空白组相比,10 ng/mL TGF-β1组、100 ng/mL TGF-β1组IL-33浓度均不同程度升高(P均<0.0083);10 ng/mL TGF-β1组IL-33浓度高于1 ng/mL TGF-β1组和100 ng/mL TGF-β1组,差异有统计学意义(P<0.0083),10 ng/mL TGF-β1组IL-33浓度升高最显著。Western blotting结果表明,空白组、1 ng/mL TGF-β1组、10 ng/mL TGF-β1组和100 ng/mL TGF-β1组细胞内IL-33表达量的总体差异有统计学意义(F=1613.0,P<0.05),各组间差异有统计学意义(P<0.0083),10 ng/mL TGF-β1组IL-33胞内表达量升高最显著。空白组、预处理TGF-β1组、未预处理SIS3组和预处理SIS3组的细胞上清IL-33浓度的总体差异有统计学意义(F=166.7,P<0.05),与空白组相比,预处理TGF-β1组、未预处理SIS3组、预处理SIS3组IL-33浓度均不同程度升高(P<0.05)。空白组、预处理TGF-β1组、未预处理SIS3组和预处理SIS3组的胞内Smad3、pSmad3、IL-33蛋白表达量的总体差异有统计学意义[(F=4752.0,P<0.05),(F=4330.0,P<0.05),(F=2791.0,P<0.05)];与空白组相比,预处理TGF-β1组、预处理SIS3组中Smad3蛋白、pSmad3蛋白、IL-33蛋白表达均不同程度升高(P<0.05),未预处理SIS3组Smad3蛋白、pSmad3蛋白、IL-33蛋白表达�Objective To investigate the signaling mechanism of interleukin(IL)-33 expression and secretion in mouse airway smooth muscle cells.Methods To observe the effect of TGF-β1 with different concentrations(0 ng/mL,1 ng/mL,10 ng/mL,100 ng/mL)on the secretion and expression of IL-33 in mouse airway smooth muscle cells.ELISA was used to detect the concentration of IL-3 in cell culture supernatant of each group.Western blotting was used to detect the expression of IL-33 protein.The inhibitory effect of TGF-β1/Smad3 signaling pathway blocker(SIS3)was observed and the cells were divided into blank group,pretreated TGF-β1 group,unpretreated SIS3 group and pretreated SIS3 group.ELISA was used to detect the concentration of IL-33 in the cell culture supernatant,and Western blotting was used to detect the expressions of Smad3,pSmad3 and IL-33 protein in each group.Results ELISA result showed that the cellular IL-33 concentration in the blank group,1 ng/mL TGF-β1 group,10 ng/mL TGF-β1 group and 100 ng/mL TGF-β1 group were statistically different(F=106.4,P<0.05).Compared with the blank group,the IL-33 concentration of 10 ng/mL TGF-β1 group and 100 ng/mL TGF-β1 group were increased(P<0.0083).The IL-33 concentration of 10 ng/mL TGF-β1 group was higher than that of 1 ng/mL TGF-β1 group and 100 ng/mL TGF-β1 group(P<0.0083).Western blotting result showed that the IL-33 protein expression in blank group,1 ng/mL TGF-β1 group,10 ng/mL TGF-β1 group and 100 ng/mL TGF-β1 group was statistically different(F=1613.0,P<0.05).There exists statiscical difference between each two groups with the most significant increase of IL-33 expression in 10 ng/mL TGF-β1 group(P<0.0083).Westerm blotting result showed that the IL-33 concentration in blank group,pretreatment TGF-β1 group,unpretreatment SIS3 group and pretreatment SIS3 group was statistically different(F=166.7,P<0.05).Compared with the blank group,the IL-33 concentration of pretreatment TGF-β1 group,unpretreatment SIS3 group and pretreatment SIS3 group were increased(P<0.05).Th

关 键 词：转化生长因子-β1/Smad3信号通路 白细胞介素-33 小鼠气道平滑肌细胞 哮喘 

分 类 号：R574[医药卫生—消化系统]