title: Hydrogen sulfide increases nitric oxide production and subsequent S-nitrosylation in endothelial cells.
abstract: Hydrogen sulfide (H2S) and nitric oxide (NO), two endogenous gaseous molecules in endothelial cells, got increased attention with respect to their protective roles in the cardiovascular system. However, the details of the signaling pathways between H2S and NO in endothelia cells remain unclear. In this study, a treatment with NaHS profoundly increased the expression and the activity of endothelial nitric oxide synthase. Elevated gaseous NO levels were observed by a novel and specific fluorescent probe, 5-amino-2-(6-hydroxy-3-oxo-3H-xanthen-9-yl)benzoic acid methyl ester (FA-OMe), and quantified by flow cytometry. Further study indicated an increase of upstream regulator for eNOS activation, AMP-activated protein kinase (AMPK), and protein kinase B (Akt). By using a biotin switch, the level of NO-mediated protein S-nitrosylation was also enhanced. However, with the addition of the NO donor, NOC-18, the expressions of cystathionine-γ-lyase, cystathionine-β-synthase, and 3-mercaptopyruvate sulfurtransferase were not changed. The level of H2S was also monitored by a new designed fluorescent probe, 4-nitro-7-thiocyanatobenz-2-oxa-1,3-diazole (NBD-SCN) with high specificity. Therefore, NO did not reciprocally increase the expression of H2S-generating enzymes and the H2S level. The present study provides an integrated insight of cellular responses to H2S and NO from protein expression to gaseous molecule generation, which indicates the upstream role of H2S in modulating NO production and protein S-nitrosylation.
authors: School of Dentistry, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
authors: Ping-Ho
authors: PH
authors: Chen
authors: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, No. 100, Shihchuan 1st Road, San Ming District, Kaohsiung 80708, Taiwan.
authors: Yaw-Syan
authors: YS
authors: Fu
authors: Department of Biological Science and Technology, Institute of Molecular Medicine and Bioengineering, National Chiao Tung University, Hsinchu 30068, Taiwan.
authors: Yun-Ming
authors: YM
authors: Wang
authors: Institute of Biotechnology, National Cheng Kung University, Tainan 70101, Taiwan.
authors: Kun-Han
authors: KH
authors: Yang
authors: Institute of Medical Science, College of Medicine, Tzu Chi University, Hualien County 97004, Taiwan.
authors: Danny Ling
authors: DL
authors: Wang
authors: Department of Biomedical Science and Environmental Biology, College of Life Science, Kaohsiung Medical University, No. 100, Shihchuan 1st Road, San Ming District, Kaohsiung 80708, Taiwan ; Department of Biological Sciences, National Sun Yat-Sen University, Kaohsiung 80424, Taiwan.
authors: Bin
authors: B
authors: Huang
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism; D002460:Cell Line; D042783:Endothelial Cells / Q000187:drug effects* / Q000378:metabolism*; D006862:Hydrogen Sulfide / Q000494:pharmacology*; D009569:Nitric Oxide / Q000096:biosynthesis*; D052250:Nitric Oxide Synthase Type III / Q000378:metabolism; D010766:Phosphorylation; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism
chemical_list: D009569:Nitric Oxide; D052250:Nitric Oxide Synthase Type III; D051057:Proto-Oncogene Proteins c-akt; D055372:AMP-Activated Protein Kinases; D006862:Hydrogen Sulfide