H2S relaxes vas deferens smooth muscle by modulating the large conductance Ca2+ -activated K+ (BKCa) channels via a redox mechanism.
BACKGROUND
Hydrogen sulfide (H(2) S) is generated in mammalian cells mainly by one of the two pyridoxal-5'-phosphate-dependent enzymes, cystathione-γ-lyase (CSE), and cystathione-β-synthase (CBS) using L-cysteine as the main substrate. In previous studies, we found that CBS and CSE were functionally expressed in vas deferens (VD) and H(2) S-mediated VD smooth muscle relaxation. However, the detail mechanisms that H(2) S-relaxed VD smooth muscle were unknown so far.


OBJECTIVE
The aim of this study is to explore the molecular target sites of H(2) S in VD smooth muscle.


METHODS
Isolated rat VD smooth muscle strips were used for tension recording in vitro. Double immunofluorescence staining was used to identify the localization of large conductance Ca(2+) -activated K(+) (BK(Ca)) channels.


METHODS
Changes in tonic contraction of isolated rat VD smooth muscle strip were measured after the treatment of drugs. The expression of BKca channels in rat VD smooth muscle cells was also assessed.


RESULTS
The results showed that L-NG-nitroarginine methyl ester (a nitric oxide synthase inhibitor) did not affect the response of VD to sodium hydrosulphide (NaHS), suggesting that nitric oxide pathway was not involved. Further studies revealed that transient receptor potential (TRP) channels did not contribute to the NaHS-induced relaxant effect. Glibenclamide, an ATP-sensitive K channel blocker, did the same thing, whereas BK(Ca) channel blockers iberiotoxin or tetraethylammonium largely reversed the relaxant effect, suggesting that H(2) S may target BK(Ca) channels. We also confirmed that BK(Ca) channels were localized in VD smooth muscle cells. Then, studies revealed that NaHS-induced VD smooth muscle relaxation was abolished by N-ethylmaleimide, which was widely used as a sulfhydryl alkylation compound protecting thiols from oxidation, whereas DL-Dithiothreitol, a strong reducing agent, did not affect the response of VD to NaHS.


CONCLUSIONS
We concluded that H(2) S relaxed the VD smooth muscle by targeting BK(Ca) channels via redox-mediated mechanism.
Department of Physiology, Shandong University School of Medicine, Jinan, China.
Yan
Y
Li
Yuanwei
Y
Zang
Shanshan
S
Fu
Hongyuan
H
Zhang
Lu
L
Gao
Jingxin
J
Li
D000818:Animals; D002478:Cells, Cultured; D006862:Hydrogen Sulfide / Q000494:pharmacology*; D066298:In Vitro Techniques; D054086:KATP Channels / Q000502:physiology; D008297:Male; D008856:Microscopy, Fluorescence; D009119:Muscle Contraction / Q000187:drug effects; D009126:Muscle Relaxation / Q000187:drug effects* / Q000502:physiology; D009129:Muscle Tonus / Q000187:drug effects / Q000502:physiology; D009130:Muscle, Smooth / Q000187:drug effects* / Q000502:physiology; D019001:Nitric Oxide Synthase / Q000502:physiology; D010084:Oxidation-Reduction; D024681:Potassium Channels, Calcium-Activated / Q000187:drug effects* / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar; D014649:Vas Deferens / Q000187:drug effects* / Q000502:physiology
D054086:KATP Channels; D024681:Potassium Channels, Calcium-Activated; D019001:Nitric Oxide Synthase; D006862:Hydrogen Sulfide