Treatment of hydrogen molecule abates oxidative stress and alleviates bone loss .pdf

太空飞行因为失重可导致航天员一系列的器官功能改变，特别是骨骼钙丢失和肌肉萎缩等，这些改变不仅对航天员的健康产生危害，而且会影响他们的运动能力，容易使他们在执行太空任务中遇到麻烦，因此开展对这些改变的机制和对抗治疗手段的研究非常重要。对一些希望正在发展太空飞行的国家尤其重要。

氢气是最近几年发现的一种非常特殊的抗氧化物质，虽然目前人们仍没有完全弄清楚氢气治疗疾病的具体分子过程和机制，但大量的临床和动物实验结果表明，氢气对许多类型的炎症、氧化损伤和细胞凋亡等具有非常显著的治疗效果，给人们试图使用氢气治疗许多重要疾病带来了许多联想。

当然民航和太空飞行还会遇到太空辐射的危害，美国航空航天局已经开展了氢气对太空辐射损伤的研究，初步研究证明氢气对太空辐射具有理想的预防效果，并提出将来用氢气作为预防太空辐射的手段。辐射损伤的最重要机制是辐射引起的自由基增加，这是自由基生物学最经典的研究结果，由于氢气具有抗氧化损伤的作用，因此使用氢气作为治疗太空飞行辐射损伤是非常容易从理论上猜测到的。

本研究者根据文献分析认为，太空飞行过程因为示众导致的骨骼肌萎缩和钙丢失和氧化损伤关系密切，采用细胞学和动物模型两类手段,证明使用氢气可以对抗失重导致的动物骨质丢失,并证明这种效应和关键信号分子例如erk、NfkB和iNOS等关系密切。由于骨质疏松不仅在太空飞行，也经常见于临床长期卧床患者的情况，长期卧床由于长期不使用肌肉运动和承受重力，非常容易发生肌肉萎缩和骨质丢失，这种情况和太空飞行存在类似性，因此本研究也提示，氢气可能对那些长期卧床患者的骨质丢失也值得尝试。曾经有研究发现，氢气对废用性肌肉萎缩有一定治疗作用，因此从这个角度考虑，氢气对那些卧床患者可能有对肌肉和骨骼的双重价值。

本研究来自北京301医院急救科，文章发表在《国际骨质疏松》杂志上。文章的题目作者和摘要如下：

Sun Y, Shuang F, Chen DM, Zhou RB. Treatment of hydrogen molecule abates oxidative stress and alleviates bone loss induced by modeled microgravity in rats. Osteoporos Int. 2012 May 31. [Epub ahead of print]

Abstract

Treatment with molecular hydrogen alleviates microgravity-induced bone loss through abating oxidative stress, restoring osteoblastic differentiation, and suppressing osteoclast differentiation and osteoclastogenesis.

INTRODUCTION:

Recently, it has been suggested that hydrogen gas exerts a therapeutic antioxidant activity by selectively reducing cytotoxic reactive oxygen species (ROS). The aim of the present study was to elucidate whether treatment with molecular hydrogen alleviated bone loss induced by modeled microgravity in rats.

METHODS:

Hindlimb suspension (HLS) and rotary wall vessel bioreactor were used to model microgravity in vivo and in vitro, respectively. Sprague-Dawley rats were exposed to HLS for 6 weeks to induced bone loss and simultaneously administrated with hydrogen water (HW). Then, we investigated the effects of incubation with hydrogen-rich medium (HRM) on MC3T3-E1 and RAW264.7 cells exposed to modeled microgravity.

RESULTS:

Treatment with HW alleviated HLS-induced reduction of bone mineral density, ultimate load, stiffness, and energy in femur and lumbar vertebra. Treatment with HW alleviated HLS-induced augmentation of malondialdehyde content and peroxynitrite content and reduction of total sulfhydryl content in femur and lumbar vertebra. In cultured MC3T3-E1 cells, incubation with HRM inhibited modeled microgravity-induced ROS formation, reduction of osteoblastic differentiation, increase of ratio of receptor activator of nuclear factor kappa B ligand to osteoprotegerin, inducible nitric oxide synthetase upregulation, and Erk1/2 phosphorylation. In cultured RAW264.7, incubation with HRM aggravated modeled microgravity-induced ROS formation, osteoclastic differentiation, and osteoclastogenesis.

CONCLUSION:

Treatment with molecular hydrogen alleviates microgravity-induced bone loss in rats. Molecular hydrogen could thus be envisaged as a nutritional countermeasure for spaceflight but remains to be tested in humans.

PMID:22648000 [PubMed - as supplied by publisher]

细胞学研究方法,日本学者是将培养瓶或板放在一个密封的湿盒内,用三个气源根据比例通气,并通过一个排气空气体排到房间外,如果先混合在灌入培养瓶,如何操作?

Over a 2-h period, we dissolved H2 into DMEM under
0.4 MPa pressure based on the method described by Ohsawa
et al. [18]. We dissolved O2 into a second medium by
bubbling O2 gas at the saturated level, and CO2 into a third
medium by bubbling CO2 gas. All three media were maintained
at atmospheric pressure. Then, we combined the three
media (H2 medium/O2 medium/CO2 medium) in the proportion
75:20:5 % (vol/vol/vol) and added fetal bovine
serum to achieve a final concentration of 1 %. For culture,
we put the combined medium into a culture flask. Then, we
filled the culture flask with mixed gas consisting of 75 %H2,
20 % O2, and 5 % CO2 (vol/vol/vol) and cultured cells in the
closed culture flask.