[1]杨娟,马莉,郭亚楠,等.lncRNA NKILA靶向miR-1236-3p对脂多糖诱导的支气管上皮细胞损伤的影响[J].第三军医大学学报,2020,42(16):1648-1654.
 YANG Juan,MA Li,GUO Yanan,et al.Long noncoding RNA NKILA alleviates lipopolysaccharide-induced bronchial epithelial cell injury by targeting miR-1236-3p[J].J Third Mil Med Univ,2020,42(16):1648-1654.
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lncRNA NKILA靶向miR-1236-3p对脂多糖诱导的支气管上皮细胞损伤的影响(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
42卷
期数:
2020年第16期
页码:
1648-1654
栏目:
基础医学
出版日期:
2020-08-30

文章信息/Info

Title:
Long noncoding RNA NKILA alleviates lipopolysaccharide-induced bronchial epithelial cell injury by targeting miR-1236-3p
作者:
杨娟马莉郭亚楠朱孟沙
河北省石家庄市第四医院儿保母乳门诊; 河北省儿童医院新生儿科; 重庆医科大学基础医学院病原生物学教研室;河北省儿童医院重症监护二科
Author(s):
YANG Juan MA Li GUO Yanan ZHU Mengsha
Children’s Healthcare and Breast Feeding Clinic, Fourth Hospital of Shijiazhuang, Shijiazhuang, Hebei Province, 050000; 2Department of Neonatology, 4Second Department of Intensive Care Medicine, Children’s Hospital of Hebei Province, Shijiazhuang, Hebei Province, 050000; 3Department of Pathogenic Biology, College of Basic Medical Sciences, Chongqing Medical University, Chongqing, 400016, China
 
关键词:
支气管上皮细胞16HBE脂多糖lncRNA NKILAmiR-1236-3p损伤凋亡
Keywords:
bronchial epithelial cells 16HBE cells lipopolysaccharide long noncoding RNA NKILA miR-1236-3p injury apoptosis
分类号:
R322.34; R363.22; R394.2
文献标志码:
A
摘要:

目的研究lncRNA NKILA对脂多糖(lipopolysaccharide,LPS)诱导的人支气管上皮细胞(human bronchial epithelial cells,16HBE)损伤的影响及潜在机制。方法用50 μg/mL LPS诱导支气管上皮细胞16HBE细胞24 h,qRT-PCR检测细胞中lncRNA NKILA和miR-1236-3p的表达。在16HBE细胞中转染pcDNA-NKILA和anti-miR-1236-3p,检测过表达lncRNA NKILA和干扰miR-1236-3p对LPS诱导的16HBE细胞损伤的影响,流式细胞术和Western blot分别检测凋亡率及凋亡相关蛋白Bcl-2和Bax的表达,ELISA法检测LPS诱导后细胞上清中白细胞介素6(interleukin 6,IL-6)、白细胞介素13(interleukin 13,IL-13)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的水平;双荧光素酶报告系统验证lncRNA NKILA与miR-1236-3p的靶向关系。结果与对照组相比,LPS诱导的支气管上皮细胞16HBE中lncRNA NKILA和抗凋亡蛋白Bcl-2含量降低,miR-1236-3p和凋亡蛋白Bax的含量升高,细胞凋亡率和炎症因子IL-6、IL-13和TNF-α含量均显著升高,均具有统计学意义(P<0.05);过表达lncRNA NKILA和干扰miR-1236-3p均可减轻LPS诱导的16HBE细胞炎症损伤,抑制细胞凋亡;lncRNA NKILA靶向负调控miR-1236-3p的表达;过表达miR-1236-3p可逆转过表达lncRNA NKILA对LPS诱导的16HBE细胞炎症和凋亡的作用。结论lncRNA NKILA通过靶向负调控miR-1236-3p的表达,减轻LPS诱导的支气管上皮细胞16HBE的炎症因子释放和凋亡。

Abstract:

ObjectiveTo investigate the protective effect of long noncoding RNA (lncRNA) NKILA against lipopolysaccharide (LPS)-induced injury of bronchial epithelial cells and explore the mechanism. MethodsBronchial epithelial cell line 16HBE was challenged with 50 μg/mL LPS for 24 h, and the changes in cellular expression levels of lncRNA NKILA and miR-1236-3p were detected using real-time quantitative PCR (qRT-PCR). 16HBE cells were transfected with pcDNA-NKILA and anti-miR-1236-3p to investigate the effects of lncRNA NKILA overexpression and miR-1236-3p interference on LPS-induced cell injury. Flow cytometry and Western blotting were used to determine the apoptosis rate and the expressions of apoptosis-related proteins Bcl-2 and Bax in the transfected cells, respectively; the levels of interleukin 6 (IL-6), IL-13, and tumor necrosis factor-α (TNF-α) in the supernatant were measured using ELISA. A dual-luciferase reporter assay was used to analyze the targeting relationship between lncRNA NKILA and miR-1236-3p. ResultsCompared with the control cells, 16HBE cells challenged with LPS showed significantly down-regulated expressions of lncRNA NKILA and Bcl-2 protein (P<0.05) and up-regulated expressions of miR-1236-3p and Bax protein (P<0.05), with significantly increased cell apoptosis rate and levels of the inflammatory factors IL-6, IL-13 and TNF-α in the supernatant (P<0.05). Over-expression of lncRNA NKILA or interference of miR-1236-3p obviously ameliorated LPS-induced inflammatory injury and inhibited apoptosis of 16HBE cells. LncRNA NKILA negatively regulated the expression of miR-1236-3p, and over-expression of miR-1236-3p reversed the effects of lncRNA NKILA over-expression on inflammation and apoptosis of LPS-treated 16HBE cells. ConclusionLncRNA NKILA reduces the release of inflammatory factors and inhibits cell apoptosis in LPS-treated 16HBE cells by targeting miR-1236-3p for negative regulation of the latter.

更新日期/Last Update: 2020-08-17