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山东大学学报 (医学版) ›› 2020, Vol. 58 ›› Issue (4): 78-83.doi: 10.6040/j.issn.1671-7554.0.2019.1345

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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

蔡秋景,张倩,何学佳,孙文丽,郭爱丽,张楠,朱薇薇   

  1. 山东大学附属济南市中心医院儿科, 山东 济南 250013
  • 发布日期:2022-09-27
  • 通讯作者: 朱薇薇. E-mail:weiweikeyan@163.com

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

CAI Qiujing, ZHANG Qian, HE Xuejia, SUN Wenli, GUO Aili, ZHANG Nan, ZHU Weiwei   

  1. Department of Pediatrics, Jinan Central Hospital Affiliated to Shandong University, Jinan 250013, Shandong, China
  • Published:2022-09-27

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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
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