Effects of uric acid in different concentrations on the proliferation of human umbilical vein endothelial cells in vitro / 中国组织工程研究

ABSTRACT

BACKGROUND:Recently, there is more and more attention on uric acid (UA), especially the correlation with cardiovascular disease in the filed of epidemiology; however, researches of the effect of UA on endothelial cells are lack based on cytology.OBJECTIVE:To explore the effects of UA in different concentrations on human umbilical vein endothelial cells line (ECV304), malondialdehyde (MDA), nitric oxide (NO) and 3-(4,5-dimethyl thiazole-2)-2, 5-diphenyl thiazolyl blue (MTT)in vitro.DESIGN: Randomized controlled study based on ECV304.5ETTING: Department of Endocrine, Xijing Hospital, the Fourth Military Medical University of Chinese PLA.MATERIALS:ECV304 was provided by Department of Immunology, the Fourth Military Medical University of Chinese PLA; DMEM Iow-glucose medium by Gibco Company, USA; bovine serum by Beijing Yuanheng Shengma Biotechnology Researching Institute; trypsin by Gibco Company, USA; MTT by Huamei Company; dimethyl sulfoxide (DMSO) analytical pure by Tianjin Bodi Chemical Engineering Co. Ltd.; UA by Sigma Company; NO kit and MDA kit by Nanjing Jiancheng Company; ordinary invert microscope and enzyme-linked immune detector IX70 invert microscope by Olympus, Japan;enzyme-linked immune detector by Eastern China Electron Tube Factory.METHODS: The experiment was carried out in Department of Endocrine and Burning, the Fourth Military Medical University of Chinese PLA from December 2003 to April 2004. Resuscitation, culture, regeneration and inoculation of endothelia cells were undertaken Cell Culture published by Situ et al. Endothelia cells were cultured with non-serum DMEM for 24hours so as to maintain synchronization at the phase of G0/G1 and divided into 4 groups, including control group, low-concentration UA group, moderate-concentration UA group and high-concentration UA group. Each group was divided into 3time points, including 24 hours, 48 hours and 72 hours with 8 samples in each time point. Samples were added with 5％serum medium containing 0 mmol/L, 0.1 mmol/L, 0.2 mmol/L and 0.4 mmol/L UA in control group, low-concentration UA group, moderate-concentration UA group and high-concentration UA group, respectively, and incubated in box with the volume fraction of 0.05 CO2 at 37 ℃. Twenty-four hours later, MDA and MTT were detected; additionally, MTT was detected once more after 48 and 72 hours.MAIN OUTCOME MEASURES: Proliferation of ECV304 and content of NO and MDA.RESULTS:With the increasing concentration of UA, content of MDA was decreased to (2.97±0.05),(2.89±0.09),(2.78±0.10) and (2.44±0.03) μmol/L, respectively; content of NO was (6.86±1.41), (12.5±2.7), (18.9±1.8) and (21.1±1.4) μ mol/L. Absorbencies of NO and MTT were increased and proliferation was increased remarkably at 48 hour.CONCLUSION: A which is characterized by anti-oxidation may promote proliferation of vascular endothelia cells and release of NO.