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

doi:10.6040/j.issn.1671-7554.0.2019.1345

摘要/Abstract

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<0.008 3);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<0.008 3),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<0.008 3),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=4 752.0,P<0.05),(F=4 330.0,P<0.05),(F=2 791.0,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蛋白表达均降低(P<0.05)。结论一定浓度的TGF-β1可刺激小鼠气道平滑肌细胞表达分泌IL-33增多,TGF-β1/Smad3信号通路可调节该过程参与哮喘。

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

Abstract: 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, 1ng/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<0.008 3). The IL-33 concentration of 10 ng/mL TGF-β1 group was higher than that of 1ng/mL TGF-β1 group and 100 ng/mL TGF-β1 group(P<0.008 3). 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=1 613.0, 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<0.008 3). 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). There were statistical difference of Smad3, pSmad3, IL-33 protein expressions among the blank group, pretreatment TGF-β1 group, unpretreatment SIS3 group and pretreatment SIS3 group［(F=4 752.0,P<0.05),(F=4 330.0,P<0.05),(F=2 791.0,P<0.05)］. Compared with the blank group, the Smad3, pSmad3 and IL-33 protein expressions were increased in pretreatment TGF-β1 group and pretreatment SIS3 group while those decreased in unpretreatment SIS3 group(P<0.05). Conclusion TGF- β1 with a certain concentration can stimulate the expression and secretion of IL-33 in mouse airway smooth muscle cells, and TGF-β1/Smad3 signaling pathway can regulate this process involved in the asthma mechanism.

Key words: Transforming growth factor-β1/Smad3 signal pathway, Interluekin-33, Mouse airway smooth muscle cell, Asthma

中图分类号:

R574

蔡秋景,张倩,何学佳,孙文丽,郭爱丽,张楠,朱薇薇. 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[J]. 山东大学学报 (医学版), 2020, 58(4): 78-83.

CAI Qiujing, ZHANG Qian, HE Xuejia, SUN Wenli, GUO Aili, ZHANG Nan, ZHU Weiwei. Airway smooth muscle cells regulate IL-33 expression through TGF-β1/Smad3 signaling pathway to participate in asthma[J]. Journal of Shandong University (Health Sciences), 2020, 58(4): 78-83.

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