基础医学与临床 ›› 2021, Vol. 41 ›› Issue (3): 333-339.

• 研究论文 • 上一篇    下一篇

miR-29b抑制LPS诱导的人支气管上皮细胞系16HBE的凋亡

袁东1,蒋瑶娜2,李亚清1   

  1. 1. 浙江省人民医院
    2. 浙江中医药大学 第二临床医学院
  • 收稿日期:2020-07-06 修回日期:2020-10-12 出版日期:2021-03-05 发布日期:2021-03-01
  • 通讯作者: 袁东 E-mail:ydq691@163.com
  • 基金资助:
    国家自然科学基金

miR-29b inhibits LPS-induced apoptosis in human bronchial epithelial cell lines 16HBE

  • Received:2020-07-06 Revised:2020-10-12 Online:2021-03-05 Published:2021-03-01

摘要: 目的 研究miR-29b对脂多糖(LPS)诱导的人支气管上皮细胞16HBE凋亡的影响。方法 将16HBE分为对照组、LPS组(50μg/mL的LPS细胞培养液培养)、转染mimics control组、转染miR-29b mimics组,MTT测定增殖,流式细胞术测定凋亡,Western blot测定c-caspase-3、c-caspase-12、糖调节蛋白78(GRP78)和CCAAT/增强子结合蛋白同源蛋白(CHOP)蛋白表达。在人支气管上皮细胞16HBE中共转染miR-29b mimics、pcDNA-CHOP,同样利用上述方法测定增殖、凋亡。结果 与对照组比较,LPS组细胞增殖活性降低,凋亡率升高,细胞中c-caspase-3、c-caspase-12、GRP78和CHOP蛋白表达水平升高。与转染mimics control组比较,转染miR-29b mimics组细胞增殖活性升高,凋亡率降低,细胞中c-caspase-3、c-caspase-12、GRP78和CHOP蛋白表达水平降低。pcDNA-CHOP可以逆转miR-29b mimics对LPS条件下支气管上皮细胞增殖和凋亡的影响。结论 miR-29b抑制LPS诱导的16HBE凋亡,机制与抑制内质网应激有关。

Abstract: Objective To study the effect of miR-29b on lipopolysaccharides(LPS)-induced human bronchial epithelial cell 16HBE apoptosis. Methods 16HBE was divided into Control group, LPS group (LPS treatment), transfection mimics control group, transfection miR-29b mimics group, MTT assay for proliferation, flow cytometry for apoptosis, and Western blot for cleaved cysteinyl aspartate specific proteinase 3(c-caspase-3), cleaved cysteinyl aspartate specific proteinase 12(c-caspase-12), Glucose tegulated protein 78(GRP78) and CCAAT/enhancer-binding protein C/EBP(CHOP)protein expression. Human bronchial epithelial cells 16HBE were co-transfected with miR-29b mimics and pcDNA-CHOP, the proliferation and apoptosis were also determined using the above method. Results Compared with the Control group, the cell proliferation activity and apoptosis rate of the LPS group decreased, and the protein expression levels of c-caspase-3, c-caspase-12, GRP78, and CHOP increased in the cells. Compared with the LPS+NC group, the LPS+miR-29b group increased cell proliferation activity, decreased apoptosis rate, and decreased c-caspase-3, c-caspase-12, GRP78, and CHOP protein expression levels. pcDNA-CHOP could reverse the effects of miR-29b mimics on the proliferation and apoptosis of bronchial epithelial cells under LPS conditions. Conclusions miR-29b inhibits 16HBE apoptosis induced by LPS, the mechanism was related to the inhibition of endoplasmic reticulum stress.