METTL3-mediated m6A methylation regulates lipopolysaccharide-induced endothelial cell permeability changes
10.19405/j.cnki.issn1000-1492.2024.06.017
- VernacularTitle:METTL3介导的m6A甲基化调控脂多糖诱导的内皮细胞通透性变化
- Author:
Jianfeng WANG
1
;
Huilin YU
;
Youxin YU
;
Junhui SONG
;
Chengying MENG
;
Wei JIANG
;
Delin HU
Author Information
1. 安徽医科大学第一附属医院烧伤科,合肥 230022
- Keywords:
sepsis;
m6A;
METTL3;
endothelial cell permeability;
lipopolysaccharide
- From:
Acta Universitatis Medicinalis Anhui
2024;59(6):1023-1028
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the molecular mechanism of N6-methyladenosine(m6A)methylation mediated by methyltransferase 3(METTL3)in regulating lipopolysaccharide(LPS)-induced endothelial cell permeability changes.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in vitro.HUVECs were treated with LPS 50,125,250,500,1 000,2 000 ng/ml for 24 h.METTL3 mRNA expression was detected by Real-time PCR.After HUVECs were intervened with 500 ng/ml for 24 h,the methylation level of m6A was detec-ted,and cell permeability was measured by cell permeability test.Real-time PCR and Western blot were used to detect mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-caherin).METTL3 overexpressed stable cell lines were constructed to measure the changes of m6A methylation level and per-meability of endothelial cells during METTL3 overexpression.Results Compared to the control group,LPS inhibi-ted the expression of HUVECs METTL3 mRNA,decreased the methylation of m6A,increased the cell permeabili-ty,and decreased the mRNA and protein expression of intercellular junction proteins(Claudin-5,Occludin and VE-Caherin).When METTL3 was overexpressed,the m6A methylation levels of endothelial cells were enhanced,and the increase of endothelial cell permeability induced by LPS was reversed.Conclusion METTL3-mediated m6A methylation can improve the permeability of endothelial cells induced by sepsis.
