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Cyanate Induces Oxidative Stress Injury in Human Umbilical Vein Endothelial Cells
Fan Chunhua1, Tian Kuan1,2, He Fei1, Hu Ling1,2, Guo Pei1,2, Chen Yi1,2, Xiong Wei1,2, Zhou Hongyu1, Li Jing1,2, Li Jinfang3, Ran Jianhua1*
1Center of Neuroscience Research, Department of Anatomy, Chongqing Medical University, Chongqing 400016, China; 2Department of Histology and Stem Cell Research, Department of Histology and Embryology, Chongqing Medical University, Chongqing 400016, China; 3Department of Neurology, the Second Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
Abstract: The study was to explore the mechanism of oxidative stress and dysfunction in human umbilical vein endothelial cells (HUVECs) treated with cyanate in vitro. The vitality of the cells treated with cyanate of different concentrations were measured by the CCK8 method. ROS level was detected by DCFH-DA assay. The NO level was detected by a commercial NO assay kit. The levels of intercelluar adhesion molecule-1 (ICAM-1) and endothelial nitric oxide synthase (eNOS) proteins were observed by immunofluorescence. Western blot analysis was used to assess the protein levels of ICAM-1 and eNOS. The activities of HUVECs in culture was inhibited by cyanate in a does dependent manner (P<0.05). Compared with the normal group and the control (mannitol) group， ROS levels were greatly increased after cyanate loading 24 h (P<0.05). NO level was decreased significantly at 24 h after cyanate treatment (P<0.05). Immunofluorescence showed high positive immunostaining of ICAM-1 after 1.00 mmol/L cyanate treatment 24 h (P<0.05). However， the immunostaining of eNOS was lowered (P<0.05). The level of ICAM-1 was remarkably increased， and eNOS was clearly reduced. Cyanate can induce in HUVECs oxidative stress and dysfunction.
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