METTL3⁃mediated m6A methylation regulates lipopolysaccharide⁃induced endothelial cell permeability changes
10.19405/j.cnki.issn1000-1492.2024.06.017
- Author:
Jianfeng Wang
1
;
Huilin Yu
2
;
Youxin Yu
1
;
Junhui Song
1
;
Chengying Meng
1
;
Wei Jiang
1
;
Delin Hu
1
Author Information
1. Dept of Burns , The First Afiliated Hospital of Anhui Medical University, Hefei 230022
2. Dept of Critical Care Medicine , Anqing First People s Hospital Afiliated to Anhui Medical University, Anqing 246003
- Publication Type:Journal Article
- 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) Ⅳinduced 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 detected , 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 permeability of endothelial cells during METTL3 overexpression.
Results :Compared to the control group , LPS inhibited the expression of HUVECs METTL3 mRNA , decreased the methylation of m6A , increased the cell permeability , 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.
- Full text:2024082011080002186METTL3介导的m6A甲...糖诱导的内皮细胞通透性变化_王建丰.pdf
