Effects of hydrogen gas on lipopolysaccharide-induced damage to human umbilical vein endothelial cells

Objective To evaluate the effects of hydrogen gas (H2) on lipopolysaccharide (LPS)-induced damage to human umbilical vein endothelial cells (HUVECs) in vitro.Methods HUVECs were cultured in DMEM culture medium containing 10％ fetal bovine serum.The cells were seeded in 6-well plates (2 ml/hole) at a density of 2 × 106 cells/ml and randomly divided into 4 groups (n =35 each) using a random number table:control group (group C),group H2,LPS group and LPS + H2 group.The cells were cultured in the plain culture medium in C and LPS groups or in hydrogen-saturated culture medium in H2 and LPS + H2 groups.In addition,LPS 1 μg/ml was added simultaneously in LPS and LPS + H2 groups,and the equal volume of normal saline was added instead in C and H2 groups.The cells were incubated for 24 h.At 6,12 and 24 h of incubation,the cells of 5 wells were chosen and stained with Wright-Giemsa to observe the destruction of HUVEC barrier.At 6,12 and 24 h of incubation,the cells of 5 wells were chosen to detect the expression of VE-cadherin and β-catenin using Western blot.At 24 h of incubation,the cells of 5 wells were chosen to detect the expression of VE-cadherin and βcatenin using immunofluorescence.Results Pathological changes of endothelial cells were observed in LPS group.The pathological changes of the cells were lighter in LPS + H2 group than in LPS group.Compared with group C,VE-cadherin expression was significantly down-regulated (P ＜ 0.05),while no significant change was found in β-catenin expression in LPS and LPS + H2 groups (P ＞ 0.05).Compared with group LPS,VE-cadherin expression was significantly up-regulated (P ＜ 0.05),while no significant change was found in ~catenin expression in group LPS + H2 (P ＞ 0.05).Conclusion H2 can effectively reduce LPS-induced damage to HUVECs through inhibiting down-regulation of VE-cadherin expression. Key words: Hydrogen ;  Lipopolysaccharides ;  Umbilical veins ;  Endothelial cells