硫化氢在大鼠骨骼肌中内源性产生，并发挥其对抗氧化应激的保护作用。

Hydrogen sulfide is endogenously generated in rat skeletal muscle and exerts a protective effect against oxidative stress.

机构信息

Department of Clinical Medicine, Capital Medical University, Beijing 100069, China.

出版信息

Chin Med J (Engl). 2013 Mar;126(5):930-6.

Abstract

BACKGROUND

Skeletal muscle has recently been recognized as an endocrine organ that can express, synthesize and secrete a variety of bioactive molecules which exert significant regulatory effects. Hydrogen sulfide (H2S) is endogenously produced in mammalian tissues and participates in a number of physiological and pathophysiological processes. We aimed to verify whether H2S could be endogenously generated and released by rat skeletal muscle, and determine the biological effects of H2S in rat skeletal muscle.

METHODS

The study was divided into two parts: detection of endogenous H2S generation and release in rat skeletal muscle and determination of antioxidative activity of skeletal muscle-derived H2S. H2S content and production in tissues were detected by sensitive sulfur electrode method. The expressions of H2S producing enzymes cystathionine β-synthase, cystathionine γ-lyase and mercaptopyruvate sulfurtransferase were detected by real-time PCR and western blotting and their tissue distributions were observed by immunohistochemical and immunofluorescent analysis. Rat skeletal muscular ischemia-reperfusion (I-R) injury model was created and evaluated by histological analysis under microscope. The malondialdehyde (MDA) contents, hydrogen peroxide levels, superoxide anion and superoxide dismutase (SOD) activities were detected using spectrophotometer.

RESULTS

H2S could be endogenously generated and released by skeletal muscle of Sprague-Dawley rats (H2S content: (2.06 ± 0.43) nmol/mg; H2S production: (0.17 ± 0.06) nmol×min(-1)×mg(-1)). Gene and protein expressions of the three H2S producing enzymes were detected in skeletal muscle, as well as the liver and kidney. Endogenous H2S content and production were decreased in skeletal muscles of rats with I-R skeletal muscle injury (P < 0.05). Furthermore, H2S significantly protected rat skeletal muscle against I-R injury and resulted in decreased MDA content, reduced hydrogen peroxide and superoxide anion levels, but increased SOD activity and protein expression in skeletal muscles (all P < 0.01).

CONCLUSION

H2S generation pathway exists in rat skeletal muscle and it acts as an antioxidant in skeletal muscle.

摘要

背景

骨骼肌最近被认为是一种内分泌器官，能够表达、合成和分泌多种具有显著调节作用的生物活性分子。内源性硫化氢（H2S）在哺乳动物组织中产生，并参与许多生理和病理生理过程。我们旨在验证 H2S 是否可以在大鼠骨骼肌中内源性产生和释放，并确定 H2S 在大鼠骨骼肌中的生物学效应。

方法

本研究分为两部分：检测大鼠骨骼肌内源性 H2S 的产生和释放，以及确定骨骼肌源性 H2S 的抗氧化活性。通过灵敏硫电极法检测组织中 H2S 的含量和产生。实时 PCR 和 Western blot 检测 H2S 产生酶胱硫醚-β-合酶、胱硫醚-γ-裂解酶和巯基丙酮酸硫转移酶的表达，并通过免疫组织化学和免疫荧光分析观察其组织分布。通过显微镜下的组织学分析来创建和评估大鼠骨骼肌缺血再灌注（I-R）损伤模型。使用分光光度计检测丙二醛（MDA）含量、过氧化氢水平、超氧阴离子和超氧化物歧化酶（SOD）活性。

结果

H2S 可以在 Sprague-Dawley 大鼠的骨骼肌中内源性产生和释放（H2S 含量：（2.06±0.43）nmol/mg；H2S 产生：（0.17±0.06）nmol×min-1×mg-1）。在骨骼肌、肝脏和肾脏中检测到三种 H2S 产生酶的基因和蛋白表达。在 I-R 骨骼肌损伤的大鼠骨骼肌中，内源性 H2S 含量和产生减少（P<0.05）。此外，H2S 显著保护大鼠骨骼肌免受 I-R 损伤，导致 MDA 含量降低，过氧化氢和超氧阴离子水平降低，而 SOD 活性和蛋白表达增加（均 P<0.01）。