Ulinastatin down regulates MiR-21 expression in murine macrophages induced by LPS and its preliminary mechanism

Xiao Ling; Ma Yu; Zhou Qiao; Zhou Fachun

doi:10.13406/j.cnki.cyxb.001245

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Ulinastatin down regulates MiR-21 expression in murine macrophages induced by LPS and its preliminary mechanism

更新时间：2024-01-17

- Ulinastatin down regulates MiR-21 expression in murine macrophages induced by LPS and its preliminary mechanism
- Journal of Chongqing Medical University Vol. 42, Issue 5, Pages: 487-493(2017)
- 作者机构：
  
  1. 重庆医科大学附属第一医院重症医学科
  2. 重庆市急救医疗中心ICU
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.001245
  CLC： R459.7
- Published：2017，
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Xiao Ling, Ma Yu, Zhou Qiao, et al. Ulinastatin down regulates MiR-21 expression in murine macrophages induced by LPS and its preliminary mechanism. [J]. Journal of Chongqing Medical University 42(5):487-493(2017)
DOI：

Xiao Ling, Ma Yu, Zhou Qiao, et al. Ulinastatin down regulates MiR-21 expression in murine macrophages induced by LPS and its preliminary mechanism. [J]. Journal of Chongqing Medical University 42(5):487-493(2017) DOI： 10.13406/j.cnki.cyxb.001245.

摘要

目的:观察在脂多糖(lipopolysaccharide

LPS)诱导的小鼠巨噬细胞株RAW264.7产生炎症反应中微小RNA-21(Micro RNA-21

mi R-21)的表达情况

以及乌司他丁(Ulinastatin

UTI)在炎症反应中对mi R-21的干预作用。方法:设置正常组、模型组(LPS 500 ng/m L剌激组)、实验组(LPS 500 ng/m L+UTI 1 000 U/m L组)和阴性实验对照组(UTI 1 000 U/m L)

观察RAW264.7细胞生长状态;四甲基偶氮唑(methyl thiazolyl tetrazolium

MTT)法检测不同浓度UTI对RAW264.7细胞活性的影响;用不同浓度UTI预处理2 h后

LPS刺激小鼠RAW264.7细胞诱导炎症模型

RT-PCR检测肿瘤坏死因子-α(tumor necrosis factor-α

TNF-α)m RNA、白介素-6(interleukin-6

IL-6)m RNA、mi R-21的表达。结果:(1)UTI对RAW264.7细胞的毒性作用:UTI浓度小于1 000 U/m L时对RAW264.7细胞无毒性作用

比较差异无统计学意义(P=0.117)。(2)与正常组相比

在不同浓度(100

250

500 ng/m L)LPS刺激RAW264.7细胞后

TNF-αm RNA及IL-6 m RNA的分泌(1.311±0.037

1.549±0.039

1.667±0.059;1.167±0.019

1.518±0.058

1.740±0.071)与基础分泌量(1.000±0.000)相比明显升高(P=0.000)

且mi R-21表达(1.082±0.114

1.671±0.087

2.789±0.107)明显升高

并随LPS浓度升高而增加(P=0.000);(3)不同浓度UTI(10

100

1 000 U/m L)能有效降低TNF-αm RNA及IL-6 m RNA(1.000±0.000

1.998±0.212

1.421±0.100

1.122±0.154

0.991±0.139;1.000±0.000

1.880±0.045

1.179±0.108

1.053±0.070

0.960±0.088)的表达

并降低mi R-21(1.000±0.000

2.789±0.107

2.675±0.135

2.258±0.170

1.369±0.279)的表达

差异有统计学意义(P1=0.000

P2=0.000

P3=0.000)。结论:LPS刺激小鼠RAW264.7细胞可促进mi R-21表达升高

其表达水平在一定程度上反映了细胞炎症反应状态;UTI可通过下调mi R-21抑制相关信号通路从而在免疫炎症反应中发挥保护作用。

Abstract

Objective:To detect the expression of micro RNA-21 in lipopolysaccharide（LPS） induced injured models of mouse RAW264.7 macrophages and the influence of Ulinastatin（UTI）on the expression of mi R-21. Methods:The growth status of RAW264.7cells was observed in the normal group

the model group（LPS）

the experimental group（LPS + UTI） and the negative control group（drug-non-stimulation group）. Methyl thiazolyl tetrazolium（MTT）assay was used to determine the effect of different concentration of UTI on the activity of RAW264.7 cells. After pre-treatment with different concentrations of UTI for 2 h

the expressions of TNF-αm RNA

IL-6 m RNA and mi R-21 were detected by RT-PCR in RAW264.7 cells induced by LPS. Results:（1）The toxic effects of UTI on RAW264.7 cell:it was non-toxic of UTI on RAW264.7 cells while the concentrations were less than 1 000 U/m L;there was no statistically significant difference（P=0.117）.（2）After RAW264.7cells being treated by LPS with different concentrations（100

250

500 ng/m L）

the expressions of mi R-21（1.000 ±0.000

1.082±0.114

1.671±0.087

2.789±0.107）and TNF-α and IL-6（1.311±0.037

1.549±0.039

1.667±0.059;1.167±0.019

1.518±0.058

1.740±0.071）were up-regulated respectively in a dose-dependent manner（P1=0.000

P2=0.000

P3=0.000）.（3）And the effect could be attenuated by UTI in different concentrations（10

100

1 000 U/m L）（mi R-21:1.000±0.000

2.789±0.107

2.675±0.135

2.258±0.170

1.369±0.279

P=0.000;TNF-α:1.000±0.000

1.998±0.212

1.421±0.100

1.122±0.154

0.991±0.139

P=0.000;IL-6:1.000±0.000

1.880±0.045

1.179±0.108

1.053±0.070

0.960±0.088

P=0.000）. Conclusion:The expression of mi R-21 is significantly changed after LPS stimulating RAW264.7macrophages

indicated that it is related to the degree of inflammation. UTI could down-regulate the mi R-21-related signaling pathway to play a protective role in immune and inflammatory response.

关键词

微小RNA-21乌司他丁炎性反应

Keywords

micro RNA-21ulinastatininflammation

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