日本学者发现,给动物饮用含氢水可以改善应激导致的神经损伤. 2008年7月19日 ... 日医大教授藉由动物实验发现喝 含氢水 可以抑制计记忆力降低, 这项对于打开认知症预防和治疗之门的成果报告在美国科学杂志电子板上发表。 这个文章是首先报道的含氢水的文献, 文章以学习记忆能力和神经干细胞为指标进行了研究. 研究比较系统,值得阅读 : Neuropsychopharmacology. 2009 Jan;34(2):501-8. Epub 2008 Jun 18. Links Consumption of molecular hydrogen prevents the stress-induced impairments in hippocampus-dependent learning tasks during chronic physical restraint in mice. Nagata K , Nakashima-Kamimura N , Mikami T , Ohsawa I , Ohta S . Department of Biochemistry and Cell Biology, Institute of Development and Aging Sciences, Graduate School of Medicine, Nippon Medical School, Nakahara-ku, Kawasaki, Japan. We have reported that hydrogen (H(2)) acts as an efficient antioxidant by gaseous rapid diffusion. When water saturated with hydrogen (hydrogen water) was placed into the stomach of a rat, hydrogen was detected at several microM level in blood. Because hydrogen gas is unsuitable for continuous consumption, we investigated using mice whether drinking hydrogen water ad libitum, instead of inhaling hydrogen gas, prevents cognitive impairment by reducing oxidative stress. Chronic physical restraint stress to mice enhanced levels of oxidative stress markers, malondialdehyde and 4-hydroxy-2-nonenal, in the brain, and impaired learning and memory, as judged by three different methods: passive avoidance learning, object recognition task, and the Morris water maze. Consumption of hydrogen water ad libitum throughout the whole period suppressed the increase in the oxidative stress markers and prevented cognitive impairment, as judged by all three methods, whereas hydrogen water did not improve cognitive ability when no stress was provided. Neural proliferation in the dentate gyrus of the hippocampus was suppressed by restraint stress, as observed by 5-bromo-2'-deoxyuridine incorporation and Ki-67 immunostaining, proliferation markers. The consumption of hydrogen water ameliorated the reduced proliferation although the mechanistic link between the hydrogen-dependent changes in neurogenesis and cognitive impairments remains unclear. Thus, continuous consumption of hydrogen water reduces oxidative stress in the brain, and prevents the stress-induced decline in learning and memory caused by chronic physical restraint. Hydrogen water may be applicable for preventive use in cognitive or other neuronal disorders 全文
这个内容说明美国这个实验室开展的工作很多,不仅对小肠而且对心脏方面,甚至在肝肺肾等方面都应该涉及. The Journal of Heart and Lung Transplantation Volume 27, Issue 2, Supplement 1 , February 2008, Page S215 Program/Abstract Issue, International Society for Heart and Lung Transplantation 28th Annual Meeting and Scientific Sessions 428: Hydrogen Inhalation Prevents Ischemia/ Reperfusion Injury of Rat Cardiac Grafts This article is not included in your organization's subscription. However, you may be able to access this article under your organization's agreement with Elsevier. A. Nakao * , 1 , 2 , D. Kaczorowski 1 , R. Sugimoto 1 , Y. Wang 1 , H. Ueda 3 , T.R. Billiar 1 and K.R. McCurry 1 , 2 1 Thomas E Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA 2 Surgery, University of Pittsburgh, Pittsburgh, PA 3 Mitleben RD Associates, Osaka, Japan Available online 25 January 2008. Article Outline Purpose Methods and Materials Results Conclusions
总体印象，美国对氢在器官移植方面非常关注，在心脏和小肠方面都已经开始出文章了。我们必须加紧这方面的工作，否则就要落后了。 该论文在实验技术上没有新东西，采用 HE 、小肠黏膜通透性、小肠体外张力功能测定，并用定量 PCR 测定了几个重要炎症分子的 mRNA 。实验非常简单。 但该文章写的比较好，基本把目前氢有关的研究都包括进，而且把 2001 年最早的一个法国的报道也用上了。尽管我早就注意到这个，但我们论文中没有引用，是我们的不足，值得学习。 该文比较可贵的是测定了动脉血中的氢浓度，呼吸后比正常升高了 3.5 倍。这个文章强调了体内细菌产生氢的量比较可观，比日本的文章要有进步。我曾经与日本学者讨论这个问题，他们说测不出来。现在的结果更明确了。 另外这个资料说明，呼吸氢并没有十分明显增加氢的量，更说明氢的作用重要，同时也提示内源性氢可能具有重要作用。文章在这个问题上提出，大肠对缺血与小肠相比不敏感，主要可能是大肠有细菌，可产生氢。内氢的作用值得我们重视。 该文在讨论中谈到， HO-1 和 IL-10 可以升高，而这些物质是抗氧化和抗炎症的。因此可以作为将来我们研究的测定指标。 在讨论中，作者认为线粒体通透性可能受到影响，可作为将来的一个指标进行研究。当然细胞凋亡相关的蛋白都是可以的。细胞的研究，很多信号分子都可以进行研究和探索。 本研究只是用 PCR 测定，没有用蛋白和酶学方法，是非常遗憾的，也是我们可以考虑在其基础上改进提高的。 Molecular hydrogen prevents intestinal I/R injury Takeshi Tsukamoto MD a , Bettina M. Buchholz MD a , Asad Nazir MD a , R. Savanh Chanthaphavong PhD a , Christopher Pape MD a , Atsunori Nakao MD a and Anthony J. Bauer PhD a a University of Pittsburgh, Pittsburgh, PA Available online 27 August 2008. Article Outline Introduction Methods Results Conclusions Introduction Molecular hydrogen shows potential for reducing ischemia/reperfusion (IR) injury. We investigate the effect of hydrogen on intestinal IR injury. Methods Rodents were subjected to SMA clamp IR (50min) and reperfusion with air or molecular hydrogen (2%) inhalation (30 pretreatment/50min ischemia/30min posttreatment). Histochemistry and IHC on jejunal muscularis whole-mounts quantified neutrophil and F4/80+ monocyte recruitment into the muscularis externa (N=4 each). Organ bath recordings measured jejunal circular muscle contractility to bethanechol (0.3-300M, N=6 each). Orally fed FITC-fluorescent microspheres (0.4M) within muscularis leukocytes assessed mucosal barrier function. Results Few neutrophils were observe in air and hydrogen treated controls (Air=1.10.23 vs. Hydrogen=0.60.14). I/R resulted in a significant increase in neutrophils into the muscularis of air treated mice (46.28.19), which was significantly decreased by hydrogen inhalation (16.74.69). Monocytes were not observed within control tissues, but I/R air mice exhibited the appearance of dense monocytic plaques at 24 hours. Hydrogen treated animals did not exhibit the monocytic plaques. Bethanechol dose-response curves demonstrated no difference between air and hydrogen treated control mice or 3hrs after reperfusion. However, 24 hours after reperfusion hydrogen treatment resulted in markedly improved muscle contractility compared to air (Air-I50min/R24hrs=0.260.071 vs. Hydrogen-I50min/R24hrs=0.680.118 g/mm2/sec @ 100M bethanechol). I/R resulted in a time dependent lumeno-lymphatic transference of microspheres in 30 minutes after reperfusion that was blocked by lymphatic ligation. Conclusions I/R injury causes a breakdown in mucosal barrier function, neutrophil and monocyte recruitment into the muscularis externa resulting in a suppression in muscle function, which is significantly prevented by molecular hydrogen. 全文
日本的论文发表后, 肯定引起了这个实验室的关注.他们课题组有日本人, 也许是通过这个途径.他们发表了1篇论文是关于移植的. 这个实验室过去一直是用呼吸一氧化碳研究其抗氧化的.看到氢的研究,很容易开展这样的工作可以理解.另外我看到两篇会议摘要也来自该实验室. 一个是关于心脏移植,一个是关于小肠缺血再灌的. 有实力的实验室可以跟踪他们的研究. 特别是他们采用呼吸的方法. 我们可以采用氢水更没有问题. 美国匹斯堡大学移植中心证明，呼吸 2% 的氢可治疗小肠移植引起的炎症损伤（ Buchholz et al. 2008 ），据说这个小组的人来中国重庆做过这个方面的报告,可惜不了解具体内容.这个文章很不错, 尽管思路没有什么新奇, 很容易想到. 我了解这个实验室曾采用饱和一氧化碳溶液研究抗氧化,估计他们肯定想到采用注射含氢水的方法. Hydrogen Inhalation Ameliorates Oxidative Stress in Transplantation Induced Intestinal Graft Injury Abstract: Ischemia/reperfusion (I/R) injury during small intestinal transplantation (SITx) frequently causes complications including dysmotility, inflammation and organ failure. Recent evidence indicates hydrogen inhalation eliminates toxic hydroxyl radicals. Syngeneic, orthotopic SITx was performed in Lewis rats with 3 h of cold ischemic time. Both donor and recipient received perioperative air or 2% hydrogen inhalation. SITx caused a delay in gastrointestinal transit and decreased jejunal circular muscle contractile activity 24 h after surgery. Hydrogen treatment resulted in significantly improved gastrointestinal transit, as well as jejunal smooth muscle contractility in response to bethanechol. The transplant induced upregulation in the inflammatory mediators CCL2, IL-1, IL-6 and TNF- were mitigated by hydrogen. Hydrogen significantly diminished lipid peroxidation compared to elevated tissue malondialdehyde levels in air-treated grafts demonstrating an antioxidant effect. Histopathological mucosal erosion and increased gut permeability indicated a breakdown in posttransplant mucosal barrier function which was significantly attenuated by hydrogen treatment. In recipient lung, hydrogen treatment also resulted in a significant abatement in inflammatory mRNA induction and reduced neutrophil recruitment. Hydrogen inhalation significantly ameliorates intestinal transplant injury and prevents remote organ inflammation via its antioxidant effects. Administration of perioperative hydrogen gas may be a potent and clinically applicable therapeutic strategy for intestinal I/R injury. 全文
国外报道 http://www.newsrx.com/article.php?articleID=1200183 日本学者 07 年 Nature Med （ Ohsawa et al. 2007 ）文章发表后，立刻引起我们的极大兴趣。我们采用不同的脑缺血模型，验证呼吸 2% 氢气是否可治疗脑缺血损伤，研究结果与日本学者的结果类似，证明呼吸 2% 氢气确实能够治疗新生儿缺血缺氧后损伤。该内容已发表文章，见 Cai JM, Kang ZM, Liu W, et al. Hydrogen therapy reduces apoptosis in neonatal hypoxia-ischemia rat model. Neurosci lett 2008,441(2):167-172. 文章发表后，立刻引起关注，目前已经被引用 6 次。 这个文章是我们的第一篇关于氢的文章, 比日本学者第一篇晚了1年. 但是我们的速度也是比较快的. Abstract: Hypoxia-ischemia (HI) brain injury is a major cause of neuronal cell death especially apoptosis in the perinatal period. This study was designated to examine the effect of hydrogen therapy on apoptosis in an established neonatal HI rat pup model. Seven-day-old rat pups were subjected to left common carotid artery ligation and then 90 min hypoxia (8% oxygen at 37 C). Immediately after HI insult, pups were placed into a chamber filled with 2% H2 for 30 min, 60 min, or 120 min, respectively. 24 hr after 2% H2 therapy, the pups were decapitated and brain injury was assessed by 2,3,5-triphenyltetrazoliumchloride (TTC), Nissl, and TUNEL staining, as well as caspase-3, caspase-12 activities in the cortex and hippocampus. H2 treatment in a duration dependent manner significantly reduced the number of positive TUNEL cells and suppressed caspase-3 and -12 activities. These results indicated H2 administration after HI appeared to provide brain protection via inhibition of neuronal apoptosis. 被引用文献目录 Ohsawa I, Nishimaki K, Yamagata K, et al. Consumption of hydrogen water prevents atherosclerosis in apolipoprotein E knockout mice.Biochem Biophys Res Commun, 2008, 377(4):1195-8 Ohta S.Hydrogen gas and hydrogen water act as a therapeutic and preventive antioxidant with a novel concept Nippon Ronen Igakkai Zasshi. 2008,45(4):355-62. Taniguchi H, Andreasson K.The hypoxic-ischemic encephalopathy model of perinatal ischemia.J Vis Exp. 2008 Nov 19;(21) pii: 955. doi: 10.3791/955 Cai JM, Kang ZM, Liu K, et al. Neuroprotective effects of hydrogen saline in neonatal hypoxia-ischemia rat model. Brain Res. 2008 Nov 27. Nakashima-Kamimura N, Mori T, Ohsawa I, Asoh S, Ohta S.Molecular hydrogen alleviates nephrotoxicity induced by an anti-cancer drug cisplatin without compromising anti-tumor activity in mice.Cancer Chemother Pharmacol. 2009 Jan 16. Gerald A. Matchett, Nancy Fathali, Yu Hasegawa, et al. Hydrogen gas is ineffective in moderate and severe neonatal hypoxia-ischemia rat models, Brain Res. 2009, doi:10.1016/j.brainres.2008.12.066 全文
我们看到日本的文章后,也进行了几个设想: 证明是否有效, 采用其他模型, 其中心肌模型也是我们想到的. 但是后来发现日本学者也有同样的思考, 文章随后发表.BBRC. 这个文章的发表让我们觉得非常遗憾. 因为我们动作不够快. Inhalation of hydrogen gas reduces infarct size in the rat model of myocardial Inhalation of hydrogen (H2) gas has been demonstrated to limit the infarct volume of brain and liver by 26 reducing ischemiareperfusion injury in rodents. When translated into clinical practice, this therapy 27 must be most frequently applied in the treatment of patients with acute myocardial infarction, since 28 angioplastic recanalization of infarct-related occluded coronary artery is routinely performed. Therefore, 29 we investigate whether H2 gas confers cardioprotection against ischemiareperfusion injury in rats. In 30 isolated perfused hearts, H2 gas enhances the recovery of left ventricular function following anoxia- 31 reoxygenation. Inhaled H2 gas is rapidly transported and can reach at risk ischemic myocardium before 32 coronary blood flow of the occluded infarct-related artery is reestablished. Inhalation of H2 gas at incom- 33 bustible levels during ischemia and reperfusion reduces infarct size without altering hemodynamic 34 parameters, thereby preventing deleterious left ventricular remodeling. Thus, inhalation of H2 gas is 35 promising strategy to alleviate ischemiareperfusion injury coincident with recanalization of coronary 36 artery 在啮齿动物实验中，吸入氢气已经被证实可以通过减轻缺血再灌注损伤而限制脑和肝的梗塞体积。当转化为临床实践时，由于血管成形术再通血管已经是常用治疗手段，这种吸入氢气限制梗塞面积的疗法将被应用于急性心肌梗死患者的治疗。因此，我们通过鼠来研究是否吸入氢气减轻缺血再灌注损伤来起到心脏保护作用。在离体灌注的心脏，氢气可以改善缺氧复氧后左室收缩功能的恢复，在梗死相关动脉的冠脉血流重建前，吸入的氢气可以迅速输送并到达处于危险的缺血心肌，缺血及再灌注期间吸入在不燃水平的氢气可以缩小梗死面积，不改变血流动力学参数，来预防心室的有害重构。因此，在冠脉血运重建时，氢气吸入将成为一种减轻缺血再灌注损伤的非常有希望的措施。 全文
日本学者随后证明呼吸2%氢可治疗肝缺血再灌注损伤(2007) 实验采取的方法十分简单, 制备肝缺血动物模型, 呼吸氢后,取组织观察HE染色确定能改善组织损伤, 测定血液中肝功能相关酶(临床常规检测手段)确定肝功能改善, 采用MDA(氧化损伤产物)确定具有抗氧化作用. 发表在 BBRC上. 尽管文章不是非常全. 如此简单的研究能发表在这个杂志上, 强烈说明这个思路的新颖性. 我看到 Nature的文章的感觉是吃惊, 看到这个文章的感觉是惊叹. 这两个文章直接引起我们强烈的兴趣: 氢真的有这样好的效果吗? 为什么? 抗氧化的研究已经有很多.氢到底神奇在什么地方? 我们随后进行了跟踪研究, 结果证明确实十分意外:效果太好了!!!!!! Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress. Kei-Ichi Fukuda, Sadamitsu Asoh, Masahiro Ishikawa, Yasuhiro Yamamoto, Ikuroh Ohsawa, Shigeo Ohta We have recently showed that molecular hydrogen has great potential for selectively reducing cytotoxic reactive oxygen species, such as hydroxyl radicals, and that inhalation of hydrogen gas decreases cerebral infarction volume by reducing oxidative stress . Here we show that the inhalation of hydrogen gas is applicable for hepatic injury caused by ischemia/reperfusion, using mice. The portal triad to the left lobe and the left middle lobe of the liver were completely occluded for 90min, followed by reperfusion for 180min. Inhalation of hydrogen gas (1-4%) during the last 190min suppressed hepatic cell death, and reduced levels of serum alanine aminotransferase and hepatic malondialdehyde. In contrast, helium gas showed no protective effect, suggesting that the protective effect by hydrogen gas is specific. Thus, we propose that inhalation of hydrogen gas is a widely applicable method to reduce oxidative stress. 全文
重新阅读日本的文章 日本的论文 , 因为有很多结果没有在正文中出现 , 都是在附件中 , 我将这些附件进行了整理 , 这样可以全面阅读该文章 . 请大家有时间要仔细阅读该全文 . 我认为有几个需要注意的问题 : 1 细胞学研究 , 采用荧光方法研究超氧阴离子和过氧化氢 , 水平都出现了下降 , 没有统计学意义 . 但是需要注意的是 , 可能已经引起了影响 , 只是检测手段不够精确 . 如果是活体连续检测 , 就不一定是这样的结果了 . 而且细胞受到损伤的情况下 , 损伤本身也影响检测效果 , 就是说细胞损伤后自由基也出现了改变 , 这种改变需要认真分析 . 到底是如何影响的 . 而且利用几个细胞来检测 , 可能存在灵敏度不足问题 自由基的测定如果能有细胞内空间分布的检测方法 , 可能更有意义 . 关于自由基的信号问题 , 我的思考是 : 从钙离子的情况来分析 , 是否自由基也存在这样的可能 , 正常自由基的水平也是一种动态的改变 , 存在震荡波 . 如果能把这个记录下来 , 将是一个非常有意思的现象 . 氢虽然不能影响其震荡频率 , 可能会影响震荡幅度 , 会影响高锋时刻引起的损伤性改变 . 2 关于正常生理指标 , 少数动物也出现一定改变 , 也需要注意 3 关于证明氢在体内被应用 , 采用动静脉血液氢水平检测 , 这个方法不准确 , 即使没有被用 , 也会出现同样的改变 , 因为气体可以有单向扩散的可能 , 就是说经过组织扩散到淋巴或者直接经过头皮扩散到空气的可能 . 证明这个的唯一方法可能是 , 对比缺血与非缺血两种情况 , 是否有不同影响 , 如果缺血的下降更明显 , 我们可以说在缺血时被用掉 . 但我认为这种减少非常少 , 不可能用这样方法检测出来 . 需要用更灵敏的方法才能实现 . 2007 年 7 月，日本医科大学学者在《自然医学》报道，动物呼吸 2% 的氢气就可有效清除自由基，显著改善脑缺血再灌注损伤，他们采用化学反应、细胞学等手段证明，氢气溶解在液体中可选择性中和羟自由基和亚硝酸阴离子，而后两者是氧化损伤的最重要介质，目前体内尚未找到内源性特异性清除途径。因此认为，氢气治疗脑缺血再灌注损伤的基础是选择性抗氧化作用（ Ohsawa et al. 2007 ）。日本学者的研究迅速引起广泛关注（已被引用 20 余次） 并引起了研究氢气治疗疾病的热潮（目前国际上已发表 12 篇相关研究论著），日本甚至专门建立了氢分子生物学研究室。随后，该研究组又用肝和心肌缺血动物模型，证明呼吸 2% 的氢气可治疗肝和心肌缺血再灌注损伤（ Fukuda et al. 2007 ； Hayashida et al. 2008 ）。采用饮用饱和氢气水可治疗应激引起的神经损伤、人类 II 型糖尿病、小鼠基因缺陷慢性氧化应激损伤和化疗药 顺铂 引起的肾损伤（ Ohsawa et al. 2008 ； Kajiyama et al. 2008 ； Sato et al. 2008 ； Nagata et al. 2009 ； Nakashima-Kamimura et al. 2009 ）。美国匹斯堡大学移植中心证明，呼吸 2% 的氢可治疗小肠移植引起的炎症损伤（ Buchholz et al. 2008 ），对小肠缺血和心脏移植后损伤同样具有保护作用（会议资料）。四川华西医院麻醉科发现，呼吸 2% 的氢可治疗肾缺血再灌注损伤（会议资料）。我们也证明，呼吸 2% 的氢可治疗新生儿脑缺血缺氧损伤（ Cai et al. 2008a ）（已被引用 6 次）。这些研究说明，作为一种选择性抗氧化物质，氢对很多疾病具有治疗作用，具有十分广泛的应用前景，也彻底推翻了氢气属于生理性惰性气体的观点。 Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals Ikuroh Ohsawa1, Masahiro Ishikawa1, Kumiko Takahashi1, Megumi Watanabe1,2, Kiyomi Nishimaki1, Kumi Yamagata1, Ken-ichiro Katsura2, Yasuo Katayama2, Sadamitsu Asoh1 Shigeo Ohta1 Acute oxidative stress induced by ischemia-reperfusion or inflammation causes serious damage to tissues, and persistent oxidative stress is accepted as one of the causes of many common diseases including cancer. We show here that hydrogen (H2) has potential as an antioxidant in preventive and therapeutic applications. We induced acute oxidative stress in cultured cells by three independent methods. H2 selectively reduced the hydroxyl radical, the most cytotoxic of reactive oxygen species (ROS), and effectively protected cells; however, H2 did not react with other ROS, which possess physiological roles. We used an acute rat model in which oxidative stress damage was induced in the brain by focal ischemia and reperfusion. The inhalation of H2 gas markedly suppressed brain injury by buffering the effects of oxidative stress. Thus H2 can be used as an effective antioxidant therapy; owing to its ability to rapidly diffuse across membranes, it can reach and react with cytotoxic ROS and thus protect against oxidative damage. 全文下: http://www.h4o.co.jp/e/pdf/Nature_070508.pdf 附件: http://www.nature.com/nm/journal/v13/n6/suppinfo/nm1577_S1.html 评论性文章: The hydrogen highway to reperfusion therapy Katherine C Wood 1 Mark T Gladwin 1 The authors are at the Vascular Medicine Branch, Intramural Research Division, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892, USA. e-mail: mgladwin@mail.nih.gov Abstract Hydrogen gas debuts as a selective antioxidant with explosive potential as cytoprotective therapy for ischemia-reperfusion injury and stroke. Top of page 111111111 评论文章
在潜水医学领域，氢氧混合气潜水过程存在呼吸数十个大气压高压氢的情况，因气体在液体中溶解量随分压增加而增加，科学家曾试图证明高压情况下，氢气或许可与氧在溶解状态下反应，或与高活性自由基发生反应，但研究并没有获得该反应存在的直接证据（ Kayar et al. 1994）。 Undersea Hyperb Med. 1994 Sep;21(3):265-75. Links Hydrogen gas is not oxidized by mammalian tissues under hyperbaric conditions. Kayar SR , Axley MJ , Homer LD , Harabin AL . Albert R. Behnke Diving Medicine Research Center, National Naval Medical Center, Bethesda, Maryland 20889-5607. Mammalian tissues, including heart, lung, liver, kidney, spleen, and skeletal muscle of guinea pig, rat, or pig, were exposed to tritium (T2) and high pressures of H2. Incorporation of the tritium label was measured to test for a latent capacity by mammalian tissues to oxidize H2 under conditions such as those experienced by deep divers breathing H2. Tissues were removed aseptically, and either minced, homogenized, or prepared as live cell cultures. The tissues were placed in a chamber to which 8 mCi T2, 1 MPa He, and either 1 or 5 MPa H2 were added. After 1 h the chamber was decompressed. The tissues were spun briefly in a vortex mixer to facilitate elimination of T2 in the gas phase. Samples were analyzed by scintillation counting for tritium incorporation in the liquid phase or in the tissues. Saline and distilled water were used as negative controls. Palladium (Pd) beads immersed in water, and cultures of the H2-metabolizing bacterium Alcaligenes eutrophus were used as positive controls. The tissues incorporated on the order of 10 nCi T2.ml-1, which implied a H2 incorporation of 10-50 nmol H2.g-1.min-1. However this incorporation was not different from that found in the water controls and was attributed to radioisotope effects. The Pd and bacterial samples incorporated over 1,000-fold more T2 than the mammalian tissues. We concluded that the mammalian tissues did not oxidize H2 under hyperbaric conditions, with a limit of detection of 100 nmol H2.g-1.min-1. 全文
法国潜水医学领域继续针对高压氢的抗氧化作用进行研究, 采用8个大气压的氢治疗肝寄生虫 曼森血吸虫感染 感染引起的肝炎症损伤,仍从抗氧化的角度去分析, 研究表明呼吸一定时间的氢具有抗炎症作用. 到这个时期, 人们考虑到氢的溶解度低,抗氧化需要比较高的分压. 思路上没有什么问题. 而且没有从选择性上考虑问题是非常遗憾的. Anti-inflammatory properties of molecular hydrogen: investigation on parasite-induced liver inflammation Bouchra Ghariba, Stphane Hannaa, Ould M.S. Abdallahia, Hubert Lepidib, Bernard Gardettec, Max De Reggia* a U399 Inserm, universit de la Mditerrane, 27, boulevard Jean-Moulin, 13005 Marseille, France b Laboratoire dhistologie, universit de la Mditerrane, 27, boulevard Jean-Moulin, 13005 Marseille, France c Comex SA, 36, boulevard des ocans,13009 Marseille, France Received 12 January 2001; accepted 25 April 2001 Communicated by Jean Rosa Abstract Molecular hydrogen reacts with the hydroxyl radical, a highly cytotoxic species produced in inflamed tissues. It has been suggested therefore to use gaseous hydrogen in a new anti-inflammatory strategy. We tested this idea, with the aid of the equipment and skills of COMEX SA in Marseille, a group who experiments with oxygenhydrogen breathing mixtures for professional deep-sea diving. The model used was schistosomiasis-associated chronic liver inflammation. Infected animals stayed 2 weeks in an hyperbaric chamber in a normal atmosphere supplemented with 0.7 MPa hydrogen. The treatment had significant protective effects towards liver injury, namely decreased fibrosis, improvement of hemodynamics, increased NOSII activity, increased antioxidant enzyme activity, decreased lipid peroxide levels and decreased circulating TNF- levels. Under the same conditions, helium exerted also some protective effects, indicating that hydroxyl radical scavenging is not the only protective mechanism. These findings indicate that the proposed anti-inflammatory strategy deserves further attention. 2001 Acadmie des sciences/ditions scientifiques et mdicales Elsevier SAS inflammation / molecular hydrogen / hydroxyl radical / hyperbaric medicine 全文下
About the establish and the administrationof Center of Molecular Hydrogen Medicine. Having Letter of Thanks from Japan Medical School byour contribution research inJapan Medical School-establish and administration Center of Molecular Hydrogen Medicine on June 25, 2008. I havebelievedin the possibility ofHydrogen Medicine.,and started this business. Ihavebeen believed in the possibilityof Hydrogen Water will bea kind ofpart inmedicine. I have been concentrated to dip anddevelophydrogen waterbusiness. It wasthankfulfor me,Japan Medical Schoolhadaccepted my contribution andhad researched hydrogen molecular medicine during 4 years. I appreciate Japan Medical Schoolhad accepted our new contribution to develop the reserch of Hydrogen Medical-in their establish and administration of Hydrogen Molecular Medicine. Hydrogen Midicineis the newest scholary research in the world. Our purpose of this contribution isto develop themedical and scholary reseach ofHydrogenMedicine. I hope Japan Medical School willmake the Center of Molecular Hydrogen Medicine to be thegreatest research center of theHydrogen Medicine in the world. June 25, 2008 Wataru Murota. President Blue Mercury, Inc. Introducing the members in the photogragh: -from the left side Ohichi Kawanami MD, Ph.D-Director, Institure of Gerontology Tsutomu ArakiMD, Ph.D-President, Nippon Medical School Wataru Murota Akiro Terashi M.D., Ph.D.-Chairman, Board of Directors of Nippon Medical School Foundation Sigeo Ohta Ph.D-Professor Ikurou Ohsawa Ph.D-Professor http://www.bluemercury.co.jp/e/Hydrogen_Medicine.html
该文首次采用注射含氢水的方法治疗脑损伤，是对去年日本Nature Med文章的改进和提高。原来的研究是采用呼吸一定浓度的氢气,达到治疗脑缺血再灌注损伤的目的，但是，通过呼吸的方法不仅在给药过程中存在爆炸的危险，而且需要比较特殊的设备，操作比较复杂，在临床上难以推广，因此，寻找更加实用的给药方法也是需要探讨的问题。经过理论推算，如果将纯氢气在生理盐水中溶解，经过一定的处理，使其达到饱和溶解，可制造出氢气的生理盐水饱和溶液，这样就可通过注射方法给药。 本文实现了这一目的. 中文摘要: 含氢气生理盐水对大鼠新生儿脑缺血的神经保护作用 脑缺血缺氧是新生儿脑损伤的重要因素。本研究目的是利用大鼠新生儿脑缺血模型，观察含氢气生理盐水对脑缺血后脑损伤的长期和短期保护作用。采用7天生大鼠，左恻颈总动脉结扎， 8%低氧37 ?C处理90分钟，制备动物模型。分别在模型制备后即刻和8小时后腹腔注射5 ml/kg饱和氢气生理盐水。24小时后断头处死动物进行TTC、尼氏和TUNEL染色。利用caspase-3活性、MDA和Iba-1免疫组化染色评价细胞死亡、炎症和氧化损伤程度。利用自发活动实验、水迷宫检测5周后神经功能情况。结果发现，氢气生理盐水对能有效降低caspase-3活性、MDA, Iba-1水平、梗死体积，提高长期神经行为功能。氢气生理盐水可能具有治疗新生儿脑缺血等各类脑缺血性疾病的价值。 本文首次证明通过注射含氢盐水可以治疗缺血损伤, 从日本首先发表呼吸2%氢治疗脑缺血的文章以后, 很多研究相继发表,证明呼吸氢可以治疗肝缺血、心肌缺血和小肠移植损伤，通过引用含氢水可以治疗人类糖尿病后氧化损伤，可以治疗应激动导致的脑损伤，可以治疗基因模型动物的脑损伤和动脉硬化。这些研究表明，给一定浓度的氢是一种非常好的治疗氧化损伤的手段。但是，通过注射的方法在临床上将是更理想的给药手段。我们经过反复研究，制备出含氢液体，并用这种注射液治疗脑损伤，效果理想。我们将在其他不同模型中继续证明这种注射液的作用，并深入研究其作用机制。我们相信，本文章的发表将会有力推动氢生物学效应的研究。希望有兴趣的战友加入我们的团队。 我们将来将希望在，缺血、炎症、创伤和各类慢性损伤模型中进行该课题研究，请继续关注我们的工作，我们相信，这些研究将给人类健康带来一个新的希望。氢的系列药物将属于高效、低毒、广谱的治疗药物。 如果将来确实能实现临床应用,本文将具有十分重要的意义. Neuroprotective Effects of Hydrogen Saline in Neonatal Hypoxia-ischemia Rat Model Abstract Cerebral hypoxiaischemia (HI) represents a major cause of brain damage in the term newborn. This study aimed to examine the short and long-term neuroprotective effect of hydrogen saline (H2 saline) using an established neonatal HI rat pup model. Seven-day-old rat pups were subjected to left common carotid artery ligation and then 90 min hypoxia (8% oxygen at 37 ?C). H2 saturated saline was administered by peritoneal injection (5 ml/kg) immediately and again at 8 h after HI insult. At 24 h after HI, the pups were decapitated and brain morphological injury was assessed by 2,3,5-triphenyltetrazolium chloride (TTC), Nissl, and TUNEL staining. Acute cell death, inflammation and oxidative stress were evaluated at 24 h by studying caspase-3 activity, MDA measurement as well as Iba-1 immunochemistry in the brain. At 5 weeks after HI, spontaneous activity test and Morris water maze test were conducted. We observed that H2 saline treatment reduced the caspase activity, MDA, Iba-1 levels, the infarct ratio, and improved the long-term neurological and neurobehavioral functions. H2 saline has potentials in the clinical treatment of HI and other ischemia-related cerebral diseases. 图片资料: 其他相关文献： 日本的第一篇文章 Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals - ?h4o.co.jp I Ohsawa, M Ishikawa, K Takahashi, M Watanabe, K - Nature Medicine, 2007 - nature.com ... 5a). Figure 5: Inhalation of hydrogen gas protects against ischemia-reperfusion injury. Figure 5 : Inhalation of hydrogen gas protects against ischemia-reperfusion injury. Rats inhaled H 2 and 30% O 2 for 1 h under the anesthetics N 2 O ... 被引用次数：14 - 相关文章 - 网页搜索 - 图书馆搜索 - 所有 5 个版本 日本的文章 Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing K Fukuda, S Asoh, M Ishikawa, Y Yamamoto, I Ohsawa - Biochemical and Biophysical Research Communications, 2007 - Elsevier We have recently showed that molecular hydrogen has great potential for selectively reducing cytotoxic reactive oxygen species, such as hydroxyl radicals, and that inhalation of hydrogen gas decreases cerebral infarction volume by ... 被引用次数：5 - 相关文章 - 网页搜索 - 图书馆搜索 - 所有 3 个版本 The hydrogen highway to reperfusion therapy - ?h4o.co.jp KC Wood, MT Gladwin - Nature Medicine, 2007 - nature.com During the ischemic phase of thromboembolic stroke, a blood clot travels to and lodges in the distal blood vessels in the brain, blocking blood flow to the oxygen-starved tissue for a period of hours. This is followed by the ... 被引用次数：1 - 相关文章 - 网页搜索 - 图书馆搜索 - 所有 4 个版本 ?Advances in Emerging Nondrug Therapies for Acute Stroke 2007 AB Singhal, EH Lo - Stroke, 2008 - ahalibrary.com From the Neuroprotection Research Laboratory, Program in Neuroscience, Harvard Medical School, and Department of Neurology, Massachusetts General Hospital, Boston, Mass. ... Numerous clinical trials of thrombolytic and ... 相关文章 - 网页快照 - 网页搜索 - 所有 5 个版本 这个也是我们的文章： Hydrogen therapy reduces apoptosis in neonatal hypoxiaischemia rat model J Cai, Z Kang, WW Liu, X Luo, S Qiang, JH Zhang, S - Neuroscience Letters, 2008 - Elsevier Hypoxiaischemia (HI) brain injury is a major cause of neuronal cell death especially apoptosis in the perinatal period. This study was designated to examine the effect of hydrogen therapy on apoptosis in an established neonatal ... 相关文章 - 网页搜索 - 图书馆搜索 - 所有 2 个版本 日本的文章 Consumption of Molecular Hydrogen Prevents the Stress-Induced Impairments in Hippocampus-Dependent K Nagata, N Nakashima-Kamimura, T Mikami, I Ohsawa - Neuropsychopharmacology, 2008 - nature.com We have reported that hydrogen (H 2 ) acts as an efficient antioxidant by gaseous rapid diffusion. When water saturated with hydrogen (hydrogen water) was placed into the stomach of a rat, hydrogen was detected at several M level in ... 相关文章 - 网页搜索 - 所有 3 个版本 日本的文章 Inhalation of hydrogen gas reduces infarct size in the rat model of myocardial ischemiareperfusion K Hayashida, M Sano, I Ohsawa, K Shinmura, K - Biochemical and Biophysical Research Communications, 2008 - Elsevier Inhalation of hydrogen (H 2 ) gas has been demonstrated to limit the infarct volume of brain and liver by reducing ischemiareperfusion injury in rodents. When translated into clinical practice, this therapy must be most ... 被引用次数：1 - 相关文章 - 网页搜索 - 图书馆搜索 - 所有 2 个版本 美国的文章 Hydrogen Inhalation Ameliorates Oxidative Stress in Transplantation Induced Intestinal Graft Injury. BM Buchholz, DJ Kaczorowski, R Sugimoto, R Yang, Y - American Journal of Transplantation, 2008 - pt.wkhealth.com ovid_logo. Search: Advanced Search. Ovid News. Learn about Ovids new research content and products. Ovid Events. Check out where Ovid is across the globe at industry conferences and events worldwide. ... 网页搜索 - 图书馆搜索 日本的文章 Hydrogen-rich pure water prevents superoxide formation in brain slices of vitamin C-depleted SMP30/ Y Sato, S Kajiyama, A Amano, Y Kondo, T Sasaki, S - Biochemical and Biophysical Research Communications, 2008 - Elsevier Hydrogen is an established anti-oxidant that prevents acute oxidative stress. To clarify the mechanism of hydrogens effect in the brain, we administered hydrogen-rich pure water (H 2 ) to senescence marker protein-30 (SMP30)/ ... 网页搜索 - 图书馆搜索 日本的文章 Inhibitory Effect of Electrolyzed Reduced Water on Tumor Angiogenesis J Ye, Y Li, T Hamasaki, N Nakamichi, T Komatsu, T - Biological Pharmaceutical Bulletin, 2008 - J-STAGE VEGF gene expression is initiated by extracellular signals in- cluding growth factors, mitogens, phorbol ester, cytokines and extracellular stresses. The first three of these exogenous signals activate the Ras-Raf-MEK-ERK ... 相关文章 - 网页搜索 - 图书馆搜索 - 所有 3 个版本 容易被遗漏的一篇早期法国文献 Anti-inflammatory properties of molecular hydrogen: investigation on parasite-induced liver B Gharib, S Hanna, OMS Abdallahi, H Lepidi, B - Comptes Rendus de l'Academie des Sciences Series III , 2001 - Elsevier ... All rights reserved. Anti-inflammatory properties of molecular hydrogen: investigation on parasite-induced liver inflammation. Premire mise en vidence des proprits anti-inflammatoires potentielles de lhydrogne molculaire. Jean Rosa. ... 被引用次数：2 - 相关文章 - 网页搜索 - 图书馆搜索 - 所有 7 个版本 文章在DXY上被好心人翻译成中文 1 引言 新生儿脑损伤最重要的原因是脑缺血缺氧，虽然其机制不是很清楚，但是神经元细胞死亡或调亡可能在其中扮演重要的作用.值得一提的是在受到损伤的最初几天内脑中半暗带发生的凋亡是可以作为治疗的靶点的，可是现在还没有治疗脑缺血缺氧病人的特异性手段。 在缺血性脑损伤包括新生儿缺血缺氧事件中炎症和氧化应激是凋亡的主要原因，有报道小神经胶质细胞参与了脑缺血缺氧诱导的炎症过程。小胶质细胞中的同种异体移植炎性因子-1是其活性升高的一个指标。缺血缺氧后的氧化应激导致DNA、细胞膜和蛋白的损伤，最终导致细胞凋亡，丙二醛是脂质胞膜氧化后的产物也是氧化损伤的标志。 有报道氢气在心、脑和肝缺血再灌注损伤后有保护作用，氢气可以中和自由基减少氧化应激，可是临床应用氢气还要考虑其安全性和方便性，本研究中，我们给予新生缺血缺氧的动物模型腹膜内注射饱和的氢气盐溶液，考察其神经保护作用。 2 结果 2.1 尼氏染色法 我们用尼染检测了氢气盐溶液的三个剂量组来找到其中的有效剂量，图1显示的是缺血缺氧24小时后的小狗脑皮层和海马的尼染结果图，发现在皮层和海马的CA1区域有大量的深色致密的神经元，而且H2W组的阳性细胞要多于模型组，我们用5ml/kg剂量组用于以下的研究，因为它的效果要好于其它剂量组。 2.2 TTC染色 图2显示了出生8天的大鼠在缺血缺氧24小时后的TTC染色结果图，发现模型组梗死区比例(10.8%)明显高于H2W组(0.99%)，后者与正常对照组相当。 2.3 丙二醛含量 检测缺血缺氧24小时后的丙二醛含量，发现模型组明显高于对照组，而给予氢气盐溶液后可以降低缺血缺氧造成的MDA含量升高。 2.4 TUNEL染色 图3是不同组的TUNEL染色放大效果图，从放大的图看，海马和皮层的细胞核可见被染色，结果显示模型组有明显增加的阳性染色，而给予氢气盐溶液后可明显可降低核染，在正常小狗的组织样本内也能见到一些核染的细胞。 2.5 细胞凋亡蛋白酶-3的活性测定 图5显示了caspase-3活性测定结果，模型组皮层caspase-3的活性为1.180.23，而海马为0.930.22。在给予氢气盐溶液后可见其活性的明显下降，结果分别为0.120.09和0.090.10，这与TUNEL染色结果相同。 2.6免疫组化分析 图4显示Iba1阳性细胞有小的核、固缩的胞质和细的分支突起，这种细胞在白质和灰质等实质中都有发现，它们的特征外形和广泛分布与经典的分支小胶质细胞都高度一致。氢气盐水处理可明显减低缺血缺氧24小时后的样品阳性细胞的数目。 2.7 体重 不同组别在处理15天后没有体重上的差异，在20天后却发现模型组和给药组体重上的差异，而对照组和给药组没有区别。 2.8 功能测试 本位反射实验印证了模型组大鼠的感觉运动功能受到影响，0分代表正常功能而1分和2分代表感觉运动功能的缺失。所有的对照组得分为0，而22.22%的模型组得分为0，77.78% 为 1，66.67%为2；63.64%给药组得分为0，36.36% 得分为1，9.09%为2。卡方检验组之间有显著的差异。 2.9 自主活动测试 为了显示行为学上的时间依赖性，图6显示了自发运动的轨迹和统计结果。结果提示所有的组大鼠在测试箱中随着时间的增加活动呈减少态势。可是给药组比模型组更活跃，移动次数也增多，而且与正常组无区别，给药组平均总距离也比模型组少。 2.10 Morris水迷宫 测定缺血缺氧24小时后大鼠的避水潜伏期，发现到达平台的时间对照组为121.23秒，而模型组为273.21秒，给予氢气盐水后时间大大缩短(151.89秒)，而且给药组与对照组无区别，提示氢气盐水提高了缺血缺氧造成的认知与学习能力的下降趋势。 3 讨论 我们考察了新生大鼠腹膜给予饱和氢气盐溶液的神经保护作用，短期作用提示了其明显减少梗死面积，增加活的神经元数目，减少凋亡细胞数量，抑制caspase-3活性，阻止小胶质细胞激活，减少氧化应激水平，这些短期的作用在缺血缺氧5周后可以转为神经保护的长期作用，提示了氢气盐溶液治疗新生儿脑功能障碍的一个潜在治疗手段。 氢气的神经保护作用是中和自由基，尤其是羟基自由基和过亚硝酸盐。羟自由基是氢氧离子的中性形式，同时也是细胞代谢的正常产物，它呈高还原性，在体内很短的半衰期（9-10秒）使其成为非常危险的基团。除此之外，细胞急剧氧化中产生了超氧负离子和一氧化氮，它们可以引起炎症反应，在正常情况下，它们一起反应并产生大量的活性氧分子过氧亚硝酸盐阴离子是潜在的氧化试剂，可引起DNA片段化与脂质氧化，同时它在体液中与二氧化碳反应形成反应中间体，后者可氧化巯基和硝酸酚类化合物，如酪氨酸。 在新生儿缺血缺氧脑损伤后，活性氧与活性氮，如OH, O2-, hydrogen dioxide (H2O2), NO, ONOO在细胞死亡中扮演重要的作用，这其中OH 和 ONOO更是强的还原性并不加区分地与核酸、脂质和蛋白反应。脑有潜在的防御体系，包括饮食中的自由基清除基（维生素C和维生素E），内源性谷胱甘肽和抗活性氧自由基酶。可是，还没有发现针对OH 和 ONOO的解毒系统。近来，Ohsawa等发现了氢气分子可选择性减少自由细胞体系中的OH and ONOO，因此在大鼠的中脑动脉闭塞模型中产生抗氧化活性的治疗作用。氢气的这一能力可能是本实验中其神经保护作用的机制。 近来研究报告显示氢气作为抗氧剂可以在脑与肝脏的缺血性损伤中通过选择性清除OH and ONOO发挥保护作用，最近一篇文献也提示了吸入氢气减少了心肌缺血再灌注损伤的大鼠模型中的梗死区面积，给予腹膜注射氢气盐溶液可以防止应激造成的身体活动受限从而导致学习记忆下降。虽然2%氢气的含量在临床上是安全的，可是实施起来并不方便，因为这需要房间或者面罩，因此，用氢气盐溶液迅速注射就不用考虑安全性的问题了。 我们选择性研究了脑损伤的两个机制：炎症和氧化应激。小胶质细胞激活可以引起炎症反应，因为在中枢神经系统内存在免疫细胞，它们对中枢性损伤如新生儿缺血缺氧起到正常的免疫应答作用并通过吞噬作用去除受损细胞。Iba1是一个147个氨基酸的钙结合蛋白，被普遍看作小胶质细胞的标志，其基因和蛋白都与AIF-1类似，后者参与各种炎症反应，如有报道其在神经性炎症紊乱和脑缺血损伤引起的小胶质细胞激活中起重要作用，其表达上调见于神经元细胞凋亡和中枢运动性神经元和感觉的受损退化。本研究中，氢气盐溶液减少脑损伤可能是通过抑制缺血缺氧造成的炎症性反应，除此之外，它也能通过减少AIF-1的表达而降低凋亡的发生，我们在另一组实验中利用氢气也得到相似的结果。氧化应激可能导致炎症的发生或者提高炎症反应，氢气盐溶液通过减轻新生儿脑中缺血缺氧造成的氧化应激就可以减少炎症和凋亡的发生。 即使氢气盐溶液和氢气本身的药代动力学有什么不同还不清楚并值得进一步研究，盐溶液优于氢气的特点就是安全性、给药的方便性和更高的溶解度，已证实高于4%的氢气浓度可导致炎症与危险，应用氢气还需要密封的房间或者面罩，以前的研究有用氢气水溶液灌胃给药的。即使口服给药是安全的但是水中的氢气容易蒸发并在胃中和小肠中损失，就很难控制浓度和吸收了，因此，我们利用腹膜注射给药就能达到迅速抽取药液迅速注射，从而避免氢气逸到周围了，氢气盐溶液的另一个比氢气和水溶液优势的就是可以达到更高的给药浓度和准确度 本研究的特点就是评价了神经生物学和行为学上的长期作用，我们发现氢气盐溶液不仅能阻止或者减少早期的病理性改变诸如梗死，或者生化改变诸如炎症和氧化应激，也能产生长期的功能恢复作用。该结果提示了其在新生儿缺血缺氧或其他脑损伤临床上的应用前景。总之，腹膜给予氢气盐溶液可以通过减轻炎症和氧化应激减少新生儿缺血缺氧所造成的脑损伤，从而减少细胞凋亡的发生，提高长期的神经生物学和行为学功能。 全文:brain res
标签: 氢气
