Protective effect of sphingosine 1-phosphate postconditioning on hypoxia/reoxygenation injury in human umbilical vein endothelial cell
10.3969/j.issn.1001-1978.2016.02.008
- VernacularTitle:1-磷酸鞘氨醇后适应对人脐静脉内皮细胞缺氧/复氧损伤的保护作用
- Author:
Mengmeng LI
;
Yuqing WANG
;
Lizhi ZHANG
;
Jianshi LOU
;
Ke WEN
- Publication Type:Journal Article
- Keywords:
S1 P;
hypoxia/reoxygenation;
HUVECs cells;
Bcl-2/Bax;
eNOS;
apoptosis
- From:
Chinese Pharmacological Bulletin
2016;(2):184-188,189
- CountryChina
- Language:Chinese
-
Abstract:
Aim To investigate the protective effects of sphingosine 1-phosphate ( S1 P ) postconditioning on hypoxia/reoxygenation( H/R) injury in human umbili-cal vein endothelial cells ( HUVEC ) and its mecha-nisms. Methods HUVECs cells were divided into five groups: normal ( control) group, S1P low concentra-tion group ( L ) , S1 P medium concentration group (M), S1P high concentration group ( H) and H/R group. MTT method was used to measure cell survival. Using flow cytometric analysis, the rate of cell apopto-sis was determined. The activities of total superoxide dismutase ( T-SOD) , copper/zinc superoxide dismuta-se ( CuZn-SOD ) , manganese superoxide dismutase ( Mn-SOD) activity, nitric oxide ( NO) and malondial-dehyde ( MDA ) content in cell culture medium were measured with colorimetry. Mitochondrial membrane potential in cells was observed with fluorescence micro-scope. Bax/Bcl-2, eNOS protein expression levels in HUVECs cells were observed with Western blot. Re-sults Compared with H/R group, S1P low, medium and high concentrations in the intervention group could significantly increase the cell survival rate after H/R injury, and increase activity of T-SOD, CuZn-SOD, Mn-SOD and decrease content of MDA. Moreover, S1 P could significantly increase NO content and in-crease eNOS protein expression, decrease apoptosis rate and inhibit the reduction of mitochondrial mem-brane potential. Conclusions S1P can decrease cell apoptosis rate of HUVECs after H/R injury with a cer-tain concentration dependence. The protection of S1P for cell apoptosis of HUVECs after H/R injury may be related to decreasing the intracellular MDA content and improving intracellular SOD activity, increasing mito-chondrial membrane potential and enhancing expres-sion of Bcl-2, anti-apoptotic protein.
