[Effect and mechanism of ubiquitin-like protein FAT10 on Angâ
¡ induced endothelial cell inflammation].
Zhonghua Xin Xue Guan Bing Za Zhi
; 51(11): 1181-1187, 2023 Nov 24.
Article
em Zh
| MEDLINE
| ID: mdl-37963754
ABSTRACT
Objective:
To investigate the role and related mechanism of ubiquitin-like protein FAT10 in the angiotensin â ¡ (Angâ ¡)-induced endothelial cell inflammatory responses.Methods:
The Western blot was used to detect the protein expression of FAT10 in 16-weeks old WKY rat carotid artery, thoracic aorta artery, renal artery and vascular smooth muscle cells (VSMC), human umbilical vein endothelial cells (HUVEC) and human breast cancer cells (MDA-MB-231). The optimal concentration and stimulation time of Angâ ¡ on inducing the highest FAT10 in HUVEC were determined. The following plasmids were constructed control plasmid, overexpression FAT10 plasmid (Flag-FAT10), invalid interference plasmid, and interference FAT10 plasmid (sh-FAT10). These plasmids were then transfected into HUVEC cells and divided into following groups control group, Flag-FAT10 group, invalid interference group, and sh-FAT10 group. After culturing with 100 nmol/L Angâ ¡ for 36 h, the control group and the Flag-FAT10 group were treated with reactive oxygen species scavenger N-acetyl-L-cysteine ââ(NAC), the protein expression levels of the inflammatory factor monocyte chemotactic protein-1 (MCP-1) and tumor necrosis factor-α (TNF-α) were measured. Laser confocal microscopy was used to detect the generation levels of reactive oxygen species in the cells of vrious groups.Results:
FAT10 was expressed in carotid artery, thoracic aorta, and renal artery of normal blood pressure rats and expressed in HUVEC, VSMC, MDA-MB-231. The expression level of FAT10 gradually increased in proportion to the increase of the time and concentration of Angâ ¡ stimulation in HUVEC, and the expression level of FAT10 was the highest when the HUVEC was treated with 100 nmol/L Angâ ¡ for 36 h (P<0.01). The protein expression level of MCP-1 (P<0.001) and TNF-α (P<0.01) was higher in Angâ ¡ treated HUVEC with FAT10 overexpression, while the expression level of MCP-1 and TNF-α protein was lower in Angâ ¡ treated HUVEC with FAT10 knockdown (all P<0.01). The level of intracellular reactive oxygen species (ROS) production was significantly increased with FAT10 overexpression (P<0.001), and the level of ROS was decreased when the expression of FAT10 was interfered (P<0.05). The increased level of MCP-1 and TNF-α proteins in FAT10 overexpressed HUVEC was reversed by NAC (all P<0.05).Conclusion:
FAT10 promotes the release of inflammatory factors induced by Angâ ¡ in endothelial cells by increasing the level of intracellular ROS production.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Angiotensina II
/
Fator de Necrose Tumoral alfa
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Zhonghua Xin Xue Guan Bing Za Zhi
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
China