[Knock-down of long intergenic noncoding RNA cyclooxygenase 2 (lincRNA-COX2) inhibits apoptosis and polarization into M1 in Listeria monocytogenes-infected macrophages].

Yurong Zhu, Shuang Huang, Lin Lin, Fengyuan Zhang, Xugan Jiang, Shengxia Chen
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Abstract

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 RAW264.7 cells were cultured and 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 MOI=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/Bcl2, 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.

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[长基因间非编码RNA环加氧酶2 (lincRNA-COX2)的敲除抑制单核细胞增生李斯特菌感染的巨噬细胞凋亡和向M1极化]。
目的探讨长基因间非编码RNA COX2 (lincRNA-COX2)对单核增生李斯特菌(Lm)感染RAW264.7细胞凋亡和细胞极化的影响。方法RAW264.7细胞培养后分为对照组(未感染细胞)、Lm感染组、小干扰RNA阴性对照(si-NC)组、si-NC和Lm感染组、lincRNA-COX2小干扰RNA (si-lincRNA-COX2)组、si-lincRNA-COX2和Lm感染组。以MOI=10 Lm感染RAW264.7细胞6小时,采用荧光显微镜和实时荧光定量PCR (qRT-PCR)检测转染siRNA的抑制效果。Western blot检测细胞内切割型caspase-3(c-caspase-3)、caspase-3、b细胞淋巴瘤-2 (Bcl2)、Bcl2相关X蛋白(BAX)、精氨酸酶1 (Arg1)、诱导型一氧化氮合酶(iNOS)的表达水平。结果与对照组相比,lm感染RAW264.7细胞c-caspase-3/caspase-3、BAX/Bcl2、iNOS水平显著上调,Arg1水平下调。LincRNA-COX2敲低抑制了lm感染RAW264.7细胞中BAX/Bcl2、c-caspase-3/caspase-3和iNOS蛋白水平的升高,而Arg1在lm感染RAW264.7细胞中表达上调。结论敲低lincRNA-COX2可抑制lm感染RAW264.7细胞的细胞凋亡,抑制巨噬细胞向M1型极化。
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