miR-20a-5p在结核分枝杆菌感染的人巨噬细胞中负向调控Jnk2基因的表达

doi:10.19871/j.cnki.xfcrbzz.2021.03.007

新发传染病电子杂志 ›› 2021, Vol. 6 ›› Issue (3): 198-201.doi: 10.19871/j.cnki.xfcrbzz.2021.03.007

miR-20a-5p在结核分枝杆菌感染的人巨噬细胞中负向调控Jnk2基因的表达

张更伟¹, 欧敏², 周雪峰³, 陶晓玉⁴, 傅佳鹏⁴, 傅向东⁴

1.深圳市第三人民医院急诊科,广东深圳 518112;
2.深圳市第三人民医院肝病研究所,广东深圳 518112;
3.广东医科大学基础医学院,广东东莞 523808;
4.深圳市第三人民医院结核病科,广东深圳 518112

收稿日期:2021-03-31 出版日期:2021-08-31 发布日期:2021-10-11
通讯作者: 傅向东,Email：fxd099@163.com;傅佳鹏,Email：zyyhlb@yeah.net
基金资助:
1.深圳市科技计划项目（JCYJ20180228162321234）; 2.深圳市卫生计生系统科研项目（SZXJ2018047）

MiR-20a-5p negatively regulates Jnk2 gene expression in human macrophages infected with Mycobacterium tuberculosis

Zhang Gengwei¹, Ou Min², Zhou Xuefeng³, Tao Xiaoyu⁴, Fu Jiapeng⁴, Fu Xiangdong⁴

1. Department of Emergency, Shenzhen Third People's Hospital, Guangdong Shenzhen 518112, China;
2. Institute of Liver Disease, Shenzhen Third People's Hospital, Guangdong Shenzhen 518112, China;
3. School of Basic Medicine, Guangdong Medical University, Guangdong Dongguan 523808, China;
4. Department of Tuberculosis, Shenzhen Third People's Hospital, Guangdong Shenzhen 518112, China

Received:2021-03-31 Online:2021-08-31 Published:2021-10-11

摘要/Abstract

摘要： 目的分析结核分枝杆菌（Mtb）感染的人巨噬细胞中miR-20a-5p对Jnk2基因表达的影响,探究miR-20a-5p基因调控细胞内信号通路的潜在机制。方法用miR-20a-5p、miR-20a-5p抑制剂（miR-20a-5p inhibitor,简写为miR-20a-inh）以及阴性对照（negative control,NC）慢病毒（慢病毒携带绿色荧光蛋白GFP标记）分别转染人巨噬细胞THP-1,流式细胞仪分选出GFP阳性THP-1细胞并用H37Ra感染,实时定量PCR技术（qPCR）检测AKT/JNK信号通路基因Akt1、Akt2、Jnk1、Jnk2的表达。蛋白质印迹法检测H37Ra感染的GFP⁺ THP-1细胞中总Jnk2和p-JNK的蛋白表达水平。结果在miR-20a-5p过表达的THP-1细胞中,Jnk2基因表达水平降低,当miR-20a-5p被抑制时,Jnk2基因表达水平升高。蛋白质印迹法检测蛋白质水平上的p-JNK2（p54）表达趋势与qPCR结果相一致,即与miR-20a-5p表达呈负相关。结论在基因和蛋白表达层面证实结核分枝杆菌感染人巨噬细胞后miR-20a-5p负向调控Jnk2表达。

关键词: 巨噬细胞, miR-20a-5p抑制剂, Jnk2基因, 人巨噬细胞THP-1, 结核分枝杆菌

Abstract: Objective To analyze the effect of miR-20a-5p on Jnk2 gene expression in human macrophages infected with Mycobacterium tuberculosis and explore the potential mechanism of mir-20a-5p gene regulating intracellular signaling pathway. Methods Human macrophages THP-1 were transfected with miR-20a-5p, miR-20a-5p-inh and negative control (NC) lentivirus, respectively. Green fluorescent protein (GFP) -positive THP-1 cells were selected by flow cytometry and infected with H37Ra. The expressions of AKT/JNK signaling pathway genes Akt1, Akt2, Jnk1 and Jnk2 were detected by qPCR.Western blotting (WB) was used to detect the expression of total Jnk2 and p-JNK in GFP⁺ THP-1 cells infected with H37Ra. Results The expression level of Jnk2 gene was decreased in THP-1 cells with miR-20a-5p overexpression, and increased when miR-20a-5p was inhibited.The expression trend of p-JNK2 (p54) at the protein level detected by WB was consistent with the qPCR results, that is, it was negatively correlated with the expression of miR-20a-5p. Conclusion miR-20a-5p negatively regulates Jnk2 expression in human macrophages infected with Mycobacterium tuberculosis at the level of gene expression and protein expression.

Key words: Macrophages, miR-20a-5p inhibitor, Jnk2 gene, Human macrophages THP-1, Mycobacterium tuberculosis

张更伟, 欧敏, 周雪峰, 陶晓玉, 傅佳鹏, 傅向东. miR-20a-5p在结核分枝杆菌感染的人巨噬细胞中负向调控Jnk2基因的表达[J]. 新发传染病电子杂志, 2021, 6(3): 198-201.

Zhang Gengwei, Ou Min, Zhou Xuefeng, Tao Xiaoyu, Fu Jiapeng, Fu Xiangdong. MiR-20a-5p negatively regulates Jnk2 gene expression in human macrophages infected with Mycobacterium tuberculosis[J]. Electronic Journal of Emerging Infectious Diseases, 2021, 6(3): 198-201.

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miR-20a-5p在结核分枝杆菌感染的人巨噬细胞中负向调控Jnk2基因的表达

MiR-20a-5p negatively regulates Jnk2 gene expression in human macrophages infected with Mycobacterium tuberculosis

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