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

杨娟; 马莉; 郭亚楠; 朱孟沙

doi:10.16016/j.1000-5404.202003257

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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

基础医学 | 更新时间：2026-01-09

- 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
- Long noncoding RNA NKILA alleviates lipopolysaccharide-induced bronchial epithelial cell injury by targeting miR-1236-3p
- 第三军医大学学报 2020年42卷第16期页码：1648-1654
- 作者机构：
  
  1.050000 石家庄，河北省石家庄市第四医院儿保母乳门诊
  2.050000 石家庄，河北省儿童医院新生儿科
  3.400016 重庆，重庆医科大学基础医学院病原生物学教研室
  4.050000 石家庄，河北省儿童医院重症监护二科
- 作者简介：
  
  [ 杨娟, E-mail: 27075915@qq.com YANG Juan, E-mail: 27075915@qq.com
- 基金信息：
  
  河北省卫生厅重点科技研究计划(20120230)
- DOI：10.16016/j.1000-5404.202003257
  中图分类号： R322.34;R363.22;R394.2
- 收稿：2020-03-20，
  
  修回：2020-05-22，
  
  纸质出版：2020-08-30
- 稿件说明：
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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.

Juan YANG, Li MA, Yanan GUO, et al. Long noncoding RNA NKILA alleviates lipopolysaccharide-induced bronchial epithelial cell injury by targeting miR-1236-3p[J]. Journal of Third Military Medical University, 2020, 42(16): 1648-1654.
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. DOI： 10.16016/j.1000-5404.202003257.

Juan YANG, Li MA, Yanan GUO, et al. Long noncoding RNA NKILA alleviates lipopolysaccharide-induced bronchial epithelial cell injury by targeting miR-1236-3p[J]. Journal of Third Military Medical University, 2020, 42(16): 1648-1654. DOI： 10.16016/j.1000-5404.202003257.

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B

ax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(

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

Objective

To investigate the protective effect of long noncoding RNA (lncRNA) NKILA against lipopolysaccharide (LPS)-induced injury of bronchial epithelial cells and explore the mechanism.

Methods

Bronchial 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.

Results

Compared with the control cells

16HBE cells challenged with LPS showed significantly down-regulated expressions of lncRNA NKILA and Bcl-2 protein (

0.05) and up-regulated expressions of miR-1236-3p and Bax protein (

0.05)

with significantly increased cell apoptosis rate and levels of the inflammatory factors IL-6

IL-13 and TNF-α in the supernatant (

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.

Conclusion

LncRNA 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.

关键词

Keywords

references

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