Oroxylin A regulates cholinergic anti-inflammatory pathway to inhibit Mycobacterium tuberculosis-induced macrophage pyroptosis
10.3760/cma.j.cn112309-20231206-00169
- VernacularTitle:千层纸素A调控胆碱能抗炎通路抑制结核分枝杆菌致巨噬细胞焦亡
- Author:
Hemin ZHANG
1
;
Jingjing SHEN
;
Fanglin LIU
;
Jianchao WU
;
Yaqi LIU
;
Xin JIANG
Author Information
1. 上海中医药大学中西医结合学院免疫学与病原生物学教研室，上海 201203
- Keywords:
Pyroptosis;
Oroxylin A;
Mycobacterium tuberculosis;
Host directly therapy;
α7 nicotinic acetylcholine receptor
- From:
Chinese Journal of Microbiology and Immunology
2024;44(5):423-430
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of oroxylin A (OrA) on macrophage pyroptosis induced by Mycobacterium tuberculosis ( Mtb) infection and the underlying molecular mechanism. Methods:An in vitro infection model was constructed by infecting J774A.1 cells with Mtb. The MTT method was used to detect the effect of OrA on the viability of J774A.1 cells. Lactate dehydrogenase (LDH) assay kit was used to detect the release of LDH by J774A.1 cells. Western blot was used to detect the protein levels of pyroptosis-related proteins such as NOD-like receptor thermal protein domain associated protein 3 (NLRP3), GSDMD-N, caspase1-p20, apoptosis-associated speck-like protein containing a CARD (ASC), and α7 nicotinic acetylcholine receptor (α7nAChR) as well as the phosphorylation of NF-κB p65. ELISA was used to detect the levels of IL-1β in each group. Immunofluorescence was performed to detect the expression and localization of α7nAChR. Results:MTT results showed that OrA had no cytotoxicity to J774A.1 cells at concentrations below 80 μmol/L. OrA reduced the release of IL-1β, down-regulated the expression of NLRP3, GSDMD-N and caspase1-p20 at protein level, inhibited ASC oligomerization, promoted the expression of α7nAChR at protein level, and inhibited the phosphorylation of NF-κB p65. In the presence of α7nAChR agonist PNU282987, the protein levels of GSDMD-N decreased and the phosphorylation of NF-κB p65 was inhibited.Conclusions:OrA may inhibits Mtb infection-induced macrophage pyroptosis through regulating cholinergic anti-inflammatory pathway.
