miR-21调控TIMP-3影响LPS诱导下RAW264.7细胞中MMP-2和MMP-9表达  

miR-21 up-regulates expression of MMP-2 and MMP-9 by targeting TIMP-3 in LPS inducing RAW264. 7 cells

作　　者：陈翔 黄莹[1] 龙春艺 廖品琥[1] CHEN Xiang;HUANG Ying;LONG Chun-Yi;LIAO Pin-Hu(Department of Respiration,Affiliated Hospital of Youjiang Medical College for Nationalities,Baise 533000,China)

机构地区：[1]右江民族医学院附属医院呼吸科

出　　处：《中国免疫学杂志》2019年第11期1295-1299,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金(No.81560321,81760350)资助

摘　　要：目的:探讨miR-21对LPS刺激下RAW264.7中基质金属蛋白酶-2(MMP-2)、MMP-9表达的影响及机制。方法:对RAW264.7细胞给予10 μg/ml LPS刺激,24 h后采用实时荧光定量PCR法和ELISA法检测miR-21和MMP-2、MMP-9在LPS组与对照组中的表达水平,并运用Pearson分析miR-21和MMP-2、MMP-9表达的相关性。构建外源性miR-21干扰的LPS刺激下RAW264.7细胞模型,采用Western blot检测miR-21对MMP-2、MMP-9和TIMP-3蛋白表达影响。结果: LPS组miR-21、MMP-2和MMP-9表达水平较对照组上升,两者呈正相关( P <0.05)。通过miR-21模拟物(miR-21 mimic)外源性上调miR-21表达,MMP-2、MMP-9表达升高,TIMP-3表达降低( P <0.05);通过miR-21抑制剂(miR-21 inhibitor)抑制miR-21表达,MMP-2、MMP-9表达降低,TIMP-3表达升高( P <0.05)。结论: miR-21能促进RAW264.7细胞MMP-2和MMP-9表达,其机制可能与靶向抑制TIMP-3有关。Objective: To discuss the influence and mechanism of miR-21 on the expression of matrix metalloproteinase-2(MMP-2)and MMP-9 stimulated by LPS in RAW264.7. Methods: RAW264.7 cells were stimulated with 10 μg/ml LPS for 24 h.The expression levels of miR-21,MMP-2 and MMP-9 in LPS group and control group were detected by real-time fluorescence quantification PCR and ELISA.Pearson correlation was used to analyze the correlation between miR-21,MMP-2 and MMP-9.RAW264.7 cell model interfered by exogenous miR-21 and stimulated by LPS was established.The effects of miR-21 on the expression of MMP-2,MMP-9 and TIMP-3 protein were detected by Western blot. Results: The expression levels of miR-21,MMP-2 and MMP-9 in LPS group were higher than those in control group,and there was a positive correlation between them( P <0.05).Exogenous up-regulation of miR-21 increased the expression of MMP-2 and MMP-9,whereas decreased TIMP-3 expression( P <0.05);on the contrary,inhibition of ed miR-21 decreased the expression of MMP-2 and MMP-9,while increased TIMP-3( P <0.05). Conclusion: miR-21 can promote the expression of MMP-2 and MMP-9 in RAW264.7 cells,and the mechanism may be related to the inhibition of targeting TIMP-3.

关 键 词：MIR-21 基质金属蛋白酶 脓毒症 RAW264.7 

分 类 号：R459.7[医药卫生—急诊医学]