title: [Regulative effects of hydrogen-rich medium on monocytic adhesion and vascular endothelial permeability].
abstract: OBJECTIVE
  To explore the regulative effects of hydrogen-rich medium on lipopolysaccharide (LPS)-induced monocytes adhesion to human umbilical vein endothelial cells (HUVEC) and vascular endothelial permeability in vitro.
  
  
  METHODS
  Endothelial cells were seeded in 6-well plates and randomly divided into 4 groups (n = 42 each):control (A), hydrogen-rich medium (B), LPS (C) and LPS+hydrogen-rich medium (D). Cells were cultured in plain culture medium in groups A and C or in hydrogen-saturated culture medium in groups B and D.LPS 1 µg/ml was added into groups C and D.When forming a monolayer, monocytes were added into each group after 6, 12 and 24 h respectively. After a 90-minute co-culturing, adhesion status was detected by Wright-Giemsa stain.Supernatants were collected to detect the concentrations of vascular cell adhesion molecule-1 (VCAM-1) and E-selectin by enzyme-linked immunosorbent assay (ELISA). The expression of VE-cadherin was measured by Western blot. Cells were stained with immunofluorescence to show the distribution of VE-cadherin after a 24-hour incubation.
  
  
  RESULTS
  Compared with group A, the adhesion of monocytes to endothelial cells increased (P < 0.05) in group C, the levels of E-selectin and VCAM-1 became elevated (P < 0.05) while the expression of VE-cadherin decreased significantly (P < 0.05). Compared with group C, adhesion decreased in group D (P < 0.05), the levels of E-selectin and VCAM-1 decreased (P < 0.05) while there was an increased expression of VE-cadherin (P < 0.05). Three timepoints showed the same tendency. The results of 24 h fluorescence indicated that, compared with group A, VE-cadherin was incomplete in cell-cell connections in group C.However it was complete and well-distributed in group D versus group C.
  
  
  CONCLUSIONS
  Hydrogen-rich medium may reduce the LPS-induced release of adhesion molecules, lessen monocytic adhesion to HUVEC and regulate the expression of VE-cadherin to protect vascular permeability.
authors.affiliation: Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Wei-na
authors.initials: WN
authors.lastname: Wang
authors.affiliation: Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Ke-liang
authors.initials: KL
authors.lastname: Xie
authors.affiliation: Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Hong-guang
authors.initials: HG
authors.lastname: Chen
authors.affiliation: Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Huan-zhi
authors.initials: HZ
authors.lastname: Han
authors.affiliation: Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China.
authors.forename: Guo-lin
authors.initials: GL
authors.lastname: Wang
authors.affiliation: Department of Anesthesiology, Tianjin Medical University General Hospital, Tianjin 300052, China. Email: yuyonghao@126.com.
authors.forename: Yong-hao
authors.initials: YH
authors.lastname: Yu
mesh_terms: D015703:Antigens, CD / Q000378:metabolism; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry*; D019040:E-Selectin / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D006859:Hydrogen / Q000494:pharmacology*; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D019010:Vascular Cell Adhesion Molecule-1 / Q000378:metabolism; D000099152:Cadherin 5
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D003470:Culture Media; D019040:E-Selectin; D006859:Hydrogen; D019010:Vascular Cell Adhesion Molecule-1; D000099152:Cadherin 5; C546936:SELE protein, human