title: Laminar shear flow increases hydrogen sulfide and activates a nitric oxide producing signaling cascade in endothelial cells.
abstract: Laminar shear flow triggers a signaling cascade that maintains the integrity of endothelial cells (ECs). Hydrogen sulfide (H2S), a new gasotransmitter is regarded as an upstream regulator of nitric oxide (NO). Whether the H2S-generating enzymes are correlated to the enzymes involved in NO production under shear flow conditions remains unclear as yet. In the present study, the cultured ECs were subjected to a constant shear flow (12 dyn/cm(2)) in a parallel flow chamber system. We investigated the expression of three key enzymes for H2S biosynthesis, cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS), and 3-mercapto-sulfurtransferase (3-MST). Shear flow markedly increased the level of 3-MST. Shear flow enhanced the production of H2S was determined by NBD-SCN reagent that can bind to cysteine/homocystein. Exogenous treatment of NaHS that can release gaseous H2S, ECs showed an increase of phosphorylation in Akt(S473), ERK(T202/Y204) and eNOS(S1177). This indicated that H2S can trigger the NO-production signaling cascade. Silencing of CSE, CBS and 3-MST genes by siRNA separately attenuated the phosphorylation levels of Akt(S473) and eNOS(S1177) under shear flow conditions. The particular mode of shear flow increased H2S production. The interplay between H2S and NO-generating enzymes were discussed in the present study.
authors.affiliation: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Center for Infectious Disease and Cancer Research, Kaohsiung Medical University, Kaohsiung 80708, Taiwan; Department of Biological Sciences, National Sun Yat-sen University, Kaohsiung 80424, Taiwan.
authors.forename: Bin
authors.initials: B
authors.lastname: Huang
authors.affiliation: Department of Chemical Engineering, National Taiwan University, Taipei 10617, Taiwan.
authors.forename: Chang-Ting
authors.initials: CT
authors.lastname: Chen
authors.affiliation: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors.forename: Chi-Shia
authors.initials: CS
authors.lastname: Chen
authors.affiliation: Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, Hsinchu 30068, Taiwan.
authors.forename: Yun-Ming
authors.initials: YM
authors.lastname: Wang
authors.affiliation: Department of Chemical Engineering, National Taiwan University, Taipei 10617, Taiwan. Electronic address: hjhsieh@ntu.edu.tw.
authors.forename: Hsyue-Jen
authors.initials: HJ
authors.lastname: Hsieh
authors.affiliation: Institute of Medical Science, College of Medicine, Tzu Chi University, Hualien County 97004, Taiwan. Electronic address: lingwang15@mail.tcu.edu.tw.
authors.forename: Danny Ling
authors.initials: DL
authors.lastname: Wang
mesh_terms: D001783:Blood Flow Velocity / Q000502:physiology*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000502:physiology*; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism*; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D009569:Nitric Oxide / Q000378:metabolism*; D033081:Shear Strength / Q000502:physiology; D013314:Stress, Mechanical
chemical_list: D009569:Nitric Oxide; D006862:Hydrogen Sulfide
keywords: 3-Mercapto-sulfurtransferase; Endothelial cell; Hydrogen sulfide; NBD-SCN; Nitric oxide; Shear flow