海南医学院学报2024,Vol.30Issue(15) :1139-1146.DOI:10.13210/j.cnki.jhmu.20240424.002

基于METTL3介导的miR-29a-3p的m6A修饰探讨平喘颗粒抑制气道上皮细胞泛凋亡治疗哮喘的机制研究

Mechanisms of Pingchuan granules in inhibiting PANoptosis of airway epithelial cells through METTL3-mediated m6A modification of miR-29a-3p in the treatment of asthma

毛旭 杨柳欣 高佳炜 王瑶 袁星星
海南医学院学报2024,Vol.30Issue(15) :1139-1146.DOI:10.13210/j.cnki.jhmu.20240424.002

基于METTL3介导的miR-29a-3p的m6A修饰探讨平喘颗粒抑制气道上皮细胞泛凋亡治疗哮喘的机制研究

Mechanisms of Pingchuan granules in inhibiting PANoptosis of airway epithelial cells through METTL3-mediated m6A modification of miR-29a-3p in the treatment of asthma

毛旭 1杨柳欣 1高佳炜 1王瑶 2袁星星3
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作者信息

  • 1. 黑龙江中医药大学,黑龙江 哈尔滨 150040
  • 2. 黑龙江中医药大学附属第一医院,黑龙江 哈尔滨 150040
  • 3. 黑龙江中医药大学,黑龙江 哈尔滨 150040;黑龙江省中医药科学院,黑龙江 哈尔滨 150006
  • 折叠

摘要

目的:观察平喘颗粒对METTL3介导的气道上皮细胞中miR-29a-3p的m6A修饰的影响,明确其抑制哮喘气道炎症的分子机制.方法:16HBE采用LPS诱导(50 mg/L)构建细胞模型,并给予平喘颗粒含药血清和地塞米松进行干预.分别采用CCK8法检测细胞活性、ELISA法检测炎症因子(TNF-α、IL-6和IL-8)的含量和miR-29a-3p的m6A修饰水平、Western blot检测METTL3与泛凋亡蛋白的表达和qRT-PCR检测METTL3与miR-29a-3p的表达.结果:与模型组相比,平喘颗粒能够显著增加16HBE的活力,抑制炎症因子TNF-α、IL-6 和IL-8 的含量,下调泛凋亡相关蛋白p-RIPK3、p-MLKL、cleaved Caspase-1、cleaved Caspase-3的表达和GSDMD-NT/FL-GSDMD与GSDME-NT/FL-GSDME的比值,差异均具有统计学意义(P<0.01).此外,平喘颗粒能够显著上调细胞中miR-29a-3p和METTL3的表达水平,促进miR-29a-3p的m6A修饰水平,与模型组相比差异均具有统计学意义(P<0.01).结论:平喘颗粒主要通过METTL3增强miR-29a-3p的m6A修饰水平,抑制气道上皮细胞泛凋亡,改善气道炎症.

Abstract

Objective:To observe the effect of Pingchuan granules on METTL3-mediated m6A modification of miR-29a-3p in airway epithelial cells,and to clarify its molecular mechanism in inhibiting airway inflammation of asthma.Methods:16HBE was induced by LPS(50 mg/L)to construct a cell model,and Pingchuan granules-containing serum and dexamethasone were given for intervention.The cell activity was detected by CCK8 assay,the content of inflammatory factors(TNF-α,IL-6 and IL-8)and the m6A modification level of miR-29a-3p were detected by ELISA,the expression of METTL3 and PAN-apoptotic protein was detected by Western blot and the expression of METTL3 and miR-29a-3p was detected by qRT-PCR,respectively.Results:Compared with the model group,Pingchuan granules could significantly increase the activity of 16HBE,inhibit the contents of in-flammatory factors TNF-α,IL-6 and IL-8,and downregulate PANoptosis-related proteins p-RIPK3,p-MLKL,cleaved Cas-pase-1,cleaved Caspase-3 and the ratio of GSDMD-NT/FL-GSDMD and GSDME-NT/FL-GSDME,the differences were sta-tistically significant(P<0.01).In addition,Pingchuan granules can significantly increase the expression levels of miR-29a-3p and METTL3 in 16HBE,and promote the m6A modification level of miR-29a-3p when compared with the model group,the differenc-es were statistically significant(P<0.01).Conclusion:Pingchuan granules enhance the m6A modification level of miR-29a-3p through METTL3,thereby inhibiting the PANoptosis of airway epithelial cells and improving airway inflammation.

关键词

平喘颗粒/支气管哮喘/泛凋亡/m6A甲基化修饰/气道上皮细胞

Key words

Pingchuan granules/Bronchial asthma/PANoptosis/M6A methylation modification/Bronchial epithelial cells

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基金项目

国家自然科学基金青年项目(82205007)

哈尔滨市科技局科研项目(2023ZCZJNS091)

出版年

2024
海南医学院学报
海南医学院

海南医学院学报

CSTPCD北大核心
影响因子:1.068
ISSN:1007-1237
参考文献量16
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