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.
School of Medicine, Jiangsu University, Zhenjiang 212013, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2023 Apr;39(4):289-294.
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.
目的 探讨长链基因间非编码RNA COX2（lincRNA-COX2）对单核细胞增生李斯特菌（Lm）感染的RAW264.7细胞凋亡及极化的影响。方法 培养RAW264.7细胞，分为对照组（未感染细胞）、Lm感染组、小干扰RNA阴性对照组（si-NC）、si-NC与Lm感染组、lincRNA-COX2小干扰RNA（si-lincRNA-COX2）组、si-lincRNA-COX2与Lm感染组。用MOI=10的Lm感染RAW264.7细胞6小时，然后通过荧光显微镜和定量实时聚合酶链反应（qRT-PCR）检测siRNA转染的抑制效率。采用蛋白质免疫印迹法检测裂解的半胱天冬酶-3（c-caspase-3）、半胱天冬酶-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感染的RAW264.7细胞中c-caspase-3/caspase-3、BAX/Bcl2和iNOS显著上调，而Arg1水平下调。敲低LincRNA-COX2可抑制Lm感染的RAW264.7细胞中BAX/Bcl2、c-caspase-3/caspase-3和iNOS蛋白水平的升高，而Lm感染的RAW264.7细胞中Arg1水平上调。结论 敲低lincRNA-COX2可抑制Lm感染的RAW264.7细胞的凋亡，并抑制巨噬细胞向M1型极化。
