乌司他丁下调LPS诱导的小鼠巨噬细胞MiR-21表达及初步机制研究

发布时间：2018-02-11 14:47

本文关键词： 微小RNA- 乌司他丁炎性反应　出处：《重庆医科大学学报》2017年05期 　论文类型：期刊论文

【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]:Aim: to observe the expression of minimal RNA-21(Micro RNA-21 in lipopolysaccharide lipopolysaccharide (LPS) -induced inflammatory reaction in murine macrophage cell line RAW264.7, and the effect of ulinastatin Ulinastatinatin UTII on the inflammatory response of murine macrophage cell line RAW264.7. Methods: normal control group was set up. The growth status of RAW264.7 cells was observed in the model group stimulated with LP500 ng/m L, the experimental group in the LP500 ng/m L UTI 1 000 U / mL group and the negative control group in the control group. The effects of different concentrations of UTI on the cell activity of RAW264.7 were detected by the method of tetrazoliumium tetrazolium tetrazolium tetrazolium (RAW264.7). After pretreatment with different concentrations of UTI for 2 h, the inflammatory model induced by RAW264.7 cells was pretreated with different concentrations of UTI. RT-PCR was used to detect the expression of tumor necrosis factor- 伪 -tumor necrosis factor- 伪 tNF- 伪 mRNAs and interleukin-6 interleukin-6m RNAi R-21. Results the toxic effect of UTI on RAW264.7 cells was less than 1 000 UMT L ~ (-1) 路min ~ (-1) ~ (-1) 路min ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1) ~ (-1). It has no toxic effect on RAW264.7 cells. There was no significant difference between the two groups.) compared with the control group, the secretion of TNF- 伪 m RNA and IL-6 m RNA in RAW264.7 cells stimulated by LPs at different concentrations was significantly higher than that in the control group (1.311 卤0.0377.49 卤0.0399.1.667 卤0.0591.167 卤0.0191.518 卤0.0581.740 卤0.0771), and the expression of R-21 was 1.082 卤0.1141.671 卤0.082.789 卤0.107), and the expression of R-21 was 1.082 卤0.1141.671 卤0.072.789 卤0.107). With the increase of LPS concentration, the expression of TNF- 伪 m RNA and IL-6 m RNA(1.000 卤0.000UmL) was significantly decreased. The difference was statistically significant (P < 0.01 0. 091 卤0. 1391.000 卤0. 1391.000 卤0. 880 卤0. 1081.179 卤0. 1081.053 卤0. 0700.60 卤0. 088)), and decreased the expression of TNF- 伪 m RNA and IL-6 m RNA(1.000 卤0. 10770 卤2. 675 卤0. 1352.258 卤0. 1700.369 卤0. 279). Conclusion LPS can stimulate the expression of TNF- 伪 RNA and IL-6 m RNA(1.000 in mice. Conclusion LPS can stimulate the expression of TNF- 伪-伪 RNA and IL-6 m RNA(1.000 in mice. Conclusion LPS can promote the expression of TNF- 伪-伪 RNA and IL-6 m RNA(1.000 in mice. Conclusion LPS can promote the expression of TNF- 伪-伪 RNA and IL-6 m RNA(1.000 + 0.1070.675 卤0.1352.258 卤0.1752.258 卤0.1700.369 卤0.2799.Conclusion LPS can promote the expression of TNF- 伪 RNA and IL-6 m RNA(1.000 in mice. To a certain extent, the expression level of UUTI may play a protective role in the immune inflammatory response by down-regulating the signal pathway associated with the inhibition of miR-21.
【作者单位】：重庆医科大学附属第一医院重症医学科;重庆市急救医疗中心ICU;
【基金】：重庆市卫生计生委科研资助项目(编号:2016ZDXM030)
【分类号】：R459.7

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