Effect of LncRNA NORAD on macrophage apoptosis via sponging miR-20a-5p in Mycobacterium tuberculosis infection
10.16718/j.1009-7708.2025.05.012
- VernacularTitle:LncRNA NORAD靶向调控miR-20a-5p对结核分枝杆菌感染的巨噬细胞凋亡的影响
- Author:
Hongmei SUN
1
;
Huanhuan CHENG
1
;
Xianglong KONG
1
;
Jianchang XUE
1
Author Information
1. 河北省胸科医院(河北省肺病重点实验室)检验科,石家庄 050041
- Publication Type:Journal Article
- Keywords:
long non-coding RNA non-coding RNA-activated by DNA damage;
microRNA-20a-5p;
Mycobacterium tuberculosis;
macrophage;
inflammation;
apoptosis
- From:
Chinese Journal of Infection and Chemotherapy
2025;25(5):549-556
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of long non-coding RNA non-coding RNA-activated by DNA damage(LncRNA NORAD)on macrophage apoptosis induced by Mycobacterium tuberculosis infection via sponging microRNA-20a-5p(miR-20a-5p).Methods Healthy subjects(n=50)who came for health checkup,patients with active tuberculosis(n=50)and individuals with asymptomatic M.tuberculosis infection(n=50)were enrolled from Hebei Chest Hospital from March 2022 to April 2023.Venous blood samples were collected to prepare serum samples.The expression levels of LncRNA NORAD,miR-20a-5p,and inflammatory factors in the serum were measured.Human monocyte line THP-1 was induced to differentiate into macrophages and assigned into Control group,Model group,transfection of NORAD empty vector group(sh-NC group),transfection of sh-NORAD vector group(sh-NORAD group),co-transfection of sh-NORAD and miR-20a-5p inhibitor empty vector group(sh-NORAD+miR-20a-5p inhibitor NC group),co-transfection of sh-NORAD and miR-20a-5p inhibitor vector group(sh-NORAD+miR-20a-5p inhibitor group),transfection of miR-20a-5p empty vector group(miR-NC group),and transfection of miR-20a-5p vector group(miR-20a-5p mimics group).The expression levels of LncRNA NORAD and miR-20a-5p(qRT PCR method),cell proliferation ability(CCK-8 kit method),cell apoptosis(flow cytometry method),inflammatory factor levels(ELISA method),and protein expression levels of BCL2-Associated X(Bax),B-cell lymphoma-2(Bcl-2),and cleaved caspase 3 in cells were detected.The targeted relationship between LncRNA NORAD and miR-20a-5p was validated.Results Compared with healthy subjects,the patients with active tuberculosis and asymptomatic M.tuberculosis infection had significantly higher serum levels of inflammatory factors and expression of LncRNA NORAD,and significantly lower miR-20a-5p.Compared with Control group,Model group had significantly higher LncRNA NORAD level,cell proliferation ability,Bcl-2 protein expression,and inflammatory factor levels,but significantly lower miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the sh-NC group,the sh-NORAD group had significantly lower LncRNA NORAD level,Bcl-2 protein expression,inflammatory factor levels,and cell proliferation ability,but significantly higher miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the sh-NORAD+miR-20a-5p inhibitor NC group,the sh-NORAD+miR-20a-5p inhibitor group had significantly higher inflammatory factor levels,Bcl-2 protein expression,and cell proliferation ability,but significantly lower miR-20a-5p level,apoptosis rate,and Bax and cleaved caspase 3 protein expression(P＜0.05).Compared with the miR-NC group,the miR-20a-5p mimics group had significantly increased inflammatory cytokines and proliferation ability,and significantly reduced apoptosis rate(P＜0.05).The targeted relationship between LncRNA NORAD and miR-20a-5p was further confirmed through experiments.Conclusions LncRNA NORAD is overexpressed in macrophages induced by M.tuberculosis.Silencing the expression of LncRNA NORAD can target the downregulation of miR-20a-5p expression,thereby inhibiting the inflammatory response of macrophages induced by M.tuberculosis and promoting cell apoptosis.
