Homocysteine induces inflammatory polarization in mouse microglia by up-regulating Rap1a
10.3969/j.issn.1000-4718.2023.12.015
- VernacularTitle:同型半胱氨酸通过上调Rap1a诱导小鼠小胶质细胞炎性极化
- Author:
Aijun WEI
1
,
2
;
Dujuan HE
;
Meikui ZHANG
Author Information
1. 陕西中医药大学第一临床医学院,陕西 咸阳 712046
2. 解放军总医院医疗保障中心远程医学科,北京 100853
- Keywords:
homocysteine;
Ras-related protein 1a;
microglia;
M1 polarization;
inflammation
- From:
Chinese Journal of Pathophysiology
2023;39(12):2242-2250
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To investigate the impact of homocysteine(Hcy)on the inflammatory response mediated by BV2 mouse microglia,and to explore the mechanism of Ras-related protein 1a(Rap1a)in the Hcy-induced inflammatory response in BV2 cells.METHODS:Mouse microglial cell line BV2 was cultured in vitro,and Hcy intervention was used to establish a hyperhomocysteinemia cellular model.The cells were divided into 4 groups:blank control group,and 50 μmol/L,100 μmol/L and 150 μmol/L Hcy groups.The mRNA expression levels of M1 polarization markers,inflammatory factors IL-6,IL-1β and TNF-α,and Rap1a in BV2 cells were detected by RT-qPCR.Additionally,the levels of inflamma-tory factors IL-6,IL-1β and TNF-α in BV2 cells were measured using an ELISA kit.The protein expression level of Rap1a was detected by Western blot assay.To verify the function of Rap1a,viral transfection was employed for both over-expression and knockdown experiments.RESULTS:Under the intervention of Hcy concentration above 100 μmol/L,BV2 cells exhibited inflammatory polarization,as indicated by the increased mRNA expression of M1 polarization markers CD80 and CD86(P＜0.05).The mRNA and protein expression levels of inflammatory factors IL-6,IL-1β and TNF-α were significantly up-regulated(P＜0.05).Additionally,the mRNA and protein expression levels of Rap1a also showed a significant increase(P＜0.05).Moreover,Rap1a mRNA level was positively correlated with CD80 mRNA,IL-1β content and TNF-α content(P＜0.05).The multiplicities of infection of the viruses with Rap1a overexpression and Rap1a knock-down were both 80,and the effective transfection was observed through fluorescence microscopy.Overexpression of Rap1a exacerbated the inflammatory polarization of BV2 cells induced by Hcy(P＜0.05),while knockdown of Rap1a attenuated this polarization(P＜0.05).CONCLUSION:Hcy can promote M1 polarization of BV2 mouse microglia,leading to in-flammatory response,which indicates that Rap1a could potentially serves as a critical regulatory factor in the Hcy-induced inflammatory response of BV2 cells.
