CD137 induces vascular muscle cells phenotype transformation through activating nuclear factor of activated T-cells 1 signaling
10.3760/cma.j.issn.0253-3758.2017.09.013
- VernacularTitle: CD137分子通过活化T细胞核因子c1调控血管平滑肌细胞表型转变
- Author:
Wei ZHONG
1
;
Bo LI
;
Jun LIU
;
Yuan XU
;
Rui CHEN
;
Chen SHAO
;
Zhongqun WANG
;
Jinchuan YAN
Author Information
1. Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China
- Publication Type:Journal Article
- Keywords:
Atherosclerosis;
Antigens, CD137;
Myocytes, smooth muscle
- From:
Chinese Journal of Cardiology
2017;45(9):799-804
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate whether CD137 induces primary vascular muscle cells (VSMCs) phenotype transformation through activating nuclear factor of activated T-cells 1(NFATc1) signaling.
Methods:VSMCs were obtained from aorta of C57BL/6J mice (8 weeks, male) through tissue-piece inoculating. Cells were divided into control group, CD137 agonist group (treated with CD137L recombinant protein) and anti-CD137 group (treated with anti-CD137 antibody). In si-RNA transfection assay, cells were divided into si-control group and si-NFATc1 group which were transfected with control or si-NFATc1 sequence respectively. The levels of NFATc1 and other phenotype related protein such as α-smooth muscle actin (α-SMA), smooth muscle myosin heavy chain (SM-MHC), vimentin were detected by Q-PCR and Western blot. Nuclear protein expression and activity of NFATc1 were detected by immunofluorescence and Western blot. Transwell assay was performed to measure the migration of VSMCs.
Results:According to Western blot, the expression of NFATc1 and vimentin was significantly upregulated (5.07±0.36 vs. 1.00±0.00, P<0.05; 3.23±0.27 vs. 1.00±0.00, P<0.05) while α-SMA and SM-MHC expressions was significantly downregulated (0.73±0.15 vs. 1.00±0.00, P<0.05; 0.45±0.05 vs. 1.00±0.00, P<0.05) in CD137 agonist group compare to control group. Compared with CD137 agonist group, the expression of NFATc1 and vimentin was significantly downregulated (1.56±0.27 vs. 5.07±0.36, P<0.05; 1.21±0.17 vs. 3.23±0.27, P<0.05), but the levels of α-SMA and SM-MHC were significantly upregulated (2.01±0.43 vs. 0.73±0.15, P<0.05; 2.85 ±0.32 vs. 0.45±0.05, P<0.05) in anti-CD137 group. Compared with si-con group, the expression of SM-MHC and α-SMA was significantly upregulated while the expression of vimentin was significantly downregulated in si-NFATc1 group. Transwell assay results demonstrated that migration cell numbers was significantly higher in CD137L group compared with control group(3.85±0.31 vs. 1.00±0.00, P<0.05), this effect was significantly attenuated by inhibiting NFATc1.
Conclusion:CD137 could induce VSMC phenotype transformation through activating NFATc1 signaling.
