摘要
目的探讨长链基因间非编码RNA环加氧酶2(lincRNA-COX2)在单核细胞增生李斯特菌(Lm)感染所致RAW264.7细胞凋亡和细胞极化中的作用.方法体外培养RAW264.7细胞,分为未感染对照组、Lm感染组、小干扰RNA阴性对照(si-NC)组、si-NC联合Lm组、lincRNA-COX2小干扰RNA(si-lincRNA-COX2)组、si-lincRNA-COX2联合Lm组.Lm感染复数(MOI)=10感染RAW264.7细胞,细菌感染6 h后收集细胞,倒置荧光显微镜和实时定量PCR检测siRNA转染抑制效率,Western blot法检测细胞中裂解型胱天蛋白酶3(c-caspase-3)、caspase-3、B细胞淋巴瘤因子2(Bcl2)、Bcl2相关X蛋白(BAX)、精氨酸酶1(Arg1)和诱导型一氧化氮合酶(iNOS)蛋白水平.结果Lm感染RAW264.7细胞后,c-caspase-3/caspase-3、BAX/Bcl2比值、iNOS蛋白水平较对照组显著上调,而Arg1水平较对照组下调.敲低lincRNA-COX2可抑制Lm感染引起的RAW264.7细胞中BAX/Bcl2、c-caspase-3/caspase-3及iNOS蛋白的升高,上调Lm感染后RAW264.7细胞中的Arg1蛋白.结论敲低lincRNA-COX2抑制Lm感染引起的RAW264.7细胞凋亡并抑制其向M1型极化.
Objective To investigate the effect of long intergenic non-coding RNA COX2(lincRNA-COX2)on apoptosis and polarization of Listeria monocytogenes(Lm)-infected RAW264.7 cells.Methods RAW264.7cellswereculturedand divided into control group(uninfected cells),Lm infection group,negative control of small interfering RNA(si-NC)group,si-NC and Lm infection group,small interfering RNA of lincRNA-COX2(si-lincRNA-COX2)group,si-lincRNA-COX2 and Lm infection group.RAW264.7 cells were infected with MOl=10 Lm for 6 hours,and then the inhibition efficiency of siRNA transfection was detected by fluorescence microscope and quantitative real-time PCR(qRT-PCR).The expression levels of cleaved-caspase-3(c-caspase-3),caspase-3,B-cell lymphoma-2(Bcl2),Bcl2 associated X protein(BAX),arginase 1(Arg1),inducible nitric oxide synthase(iNOs)were detected by Western blot analysis.Results c-caspase-3/caspase-3,BAX/Bcl2 and iNOS were significantly up-regulated,while the level of Arg1 was down-regulated in Lm-infected RAW264.7 cells compared with control group.LincRNA-COX2 knockdown inhibited the increase of protein levels for BAX/Bch2,c-caspase-3/caspase-3 and iNOS in Lm-infected RAW264.7 cells,while the level of Arg1 in Lm-infected RAW264.7 cells was up-regulated.Conclusion Knockdown of lincRNA-COX2 can inhibit cell apoptosis and suppress the macrophage polarization into M1 type in Lm-infected RAW264.7 cells.
作者
朱昱蓉
黄霜
林琳
张峰源
姜旭淦
陈盛霞
ZHU Yurong;HUANG Shuang;LIN Lin;ZHANG Fengyuan;JIANG Xugan;CHEN Shengxia(School of Medicine,Jiangsu University,Zhenjiang 212013;Clinical Microbiology,Linfen Central Hospital,Linfen 041000,China)
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2023年第4期289-294,共6页
Chinese Journal of Cellular and Molecular Immunology
基金
中国科技部基础平台项目(2019-194-30)。
