Changes of connexin 43 in irradiated human vascular endothelial cells and its influence on cell stiffness
10.3760/cma.j.issn.0254-5098.2021.06.004
- VernacularTitle:缝隙连接蛋白43在受照人血管内皮细胞中的改变及其对受照细胞刚性的作用
- Author:
Chen LI
1
;
Mei TIAN
;
Qiao GOU
;
Xuesong QI
;
Xu SU
Author Information
1. 中国疾病预防控制中心辐射防护与核安全医学所 辐射防护与核应急中国疾病预防控制中心重点实验室，北京 100088
- Keywords:
X-rays;
HUVEC cell;
Cx43;
Stiffness
- From:
Chinese Journal of Radiological Medicine and Protection
2021;41(6):418-425
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the changes of connexin 43 (Cx43) in human umbilical vein endothelial cells (HUVEC) after X-ray irradiation and its influence on the stiffness of irradiated cells.Methods:Western blot was used to detect the expression of Cx43 in HUVEC cells at different time points (0, 6, 12, 24 and 48 h) after different doses of X-ray irradiation (0, 2.5, 5, 10 and 20 Gy), and the phosphorylation levels of three phosphorylation sites (Ser279/282, Ser368 and Tyr265) of Cx43 at different time points (3, 6, 24 and 48 h) after 0, 5 and 10 Gy irradiation. The distribution of Cx43 protein in the irradiated HUVEC cells was detected by immunofluorescence. The stiffness changes of cells were detected by atomic force microscopy (AFM) at the depths of 50, 100 and 200 nm.Results:The expression of Cx43 in HUVEC cells was reduced at 6, 12, 24 and 48 h after 10 Gy X-ray irradiation( t=3.262, 3.708, 3.686, 6.825, P<0.05)and this decrease had a dose dependent manner at 24 h after 2.5, 5, 10 and 20 Gy irradiation ( t=3.034, 10.720, 13.130, 13.650, P<0.05). At 24 h after 5, 10 and 20 Gy X-ray irradiation, the distribution of Cx43 in HUVEC cells was transported from intercellular gap junctions to nucleus and perinuclear region. At 24-48 h after irradiation, the phosphorylation level of Ser368 at Cx43 increased and in a dose dependent manner. At 24 h after irradiation, the stiffness of the irradiated cells decreased significantly under the conditions of 100 and 200 nm ( t=3.362, 5.122, P<0.05), and recovered with overexpression of Cx43 ( t=2.674, 4.398, P<0.05). Conclusions:X-ray irradiation leads to the phosphorylation of Ser368 at Cx43, which promotes the degradation and nucleus/perinuclear translocation of Cx43 and reduces the stiffness of HUVEC. Increasing the expression level of Cx43 is helpful to the stiffness recovery of irradiated vascular endothelial cells, suggesting that Cx43 may be a potential target for regulating radiation injury of vascular endothelial cells.
