氢水对小肠缺血再灌注损伤的保护作用早有研究，本研究的出发点是认为通过腹腔和静脉注射可能无法保证小肠局部药物浓度，通过不同给药方法对局部氢浓度效果的比较，认为口服才是最理想的给氢水方法。这个思路的前提是，氢浓度越高效果越理想，其实这个基础并不是非常坚固，因为过去和现在的研究都没有剂量效应的比较。不过这是研究的基础，从逻辑上是可行的。在检测局部氢气浓度的基础上，作者选择了口服给氢水的方法，观察了对小肠缺血后组织学、细胞凋亡数量、氧化损伤和炎症因子等的影响，确定了这种方法可以作为小肠缺血保护的手段，这一研究目的是将这种手段作为小肠再体保护的方法，作为小肠移植一种小肠保护策略。

研究中采用组织气体挥发，气相色谱法检测氢气浓度来估计局部组织氢气浓度的变化规律，值得推荐，这比采用电极法的优点是稳定性好，能获得比较准确的数据，缺点是难以进行连续检测，无法获得准确全面的氢气变化规律。不过检测的准确度是第一位的，不能为了收集全面的信息丧失准确检测这一目标。

氢水技术来自日本Miz公司，这一公司给日本学术研究提供了大量研究用氢水，核心是能现场制备出氢气浓度很高的产品，这是许多日本学者放弃使用市场上的包装氢气水而选择该产品的重要原因。日本市场上现在供应的多种品牌的氢水，存在的最大问题是氢气浓度非常低，而且不稳定，这不仅对消费者不公平，也对学术研究带来不利影响。国内相关产品制造者应该引以为戒，在氢气浓度和稳定性这方面一定引起足够重视。千万不能只借用概念，不注重质量，这样不可能长期成功，而且对整个领域有危害。只有基础坚实了，才能最终明确效果，甚至效果的程度。

氢气浓度检测方法：

Approximately 1 g of tissue sample was obtained and put into a small container (24 mL) for the measurement of hydrogen concentration. The hydrogen concentration was measured using a sensor gas chromatograph, SGHA-PA (FIS Inc., Hyogo, Japan) after pulverizing the tissue using gentleMACS Octo Dissociator (Miltenyi biotec GmbH, Bergisch Gladbach, Germany). Hydrogen concentration of tissue samples was measured after cleaning tissue samples with saline to avoid mixture of hydrogen rich solution into a small container. Hydrogen concentration in 24 mL container was converted into concentration per 1 mL.

Transplantation. 2014 Dec 23. [Epub ahead of print]

Luminal Injection of Hydrogen-Rich Solution Attenuates Intestinal Ischemia-Reperfusion Injury in Rats.

Shigeta T1, Sakamoto S, Li X, Cai S, Liu C, Kurokawa R, Nakazawa A, Kasahara M, Uemoto S.

Author information11 Organ Transplant Center, National Center for Child Health and Development, Tokyo, Japan.

BACKGROUND: Luminal preservation of the intestine is an attractive method to locally mitigate preservation injury and ischemic-reperfusion injury in small bowel transplantation (SBT) because this method has a potential to maintain the intestinal graft integrity. Hydrogen is noted as an antioxidant material by reducing hydroxyl radicals. We hypothesized that hydrogen-containing solution can be an optimum material for luminal preservation method in SBT.

METHODS: Ischemic reperfusion was induced in Lewis rats by occlusion of the supramesenteric artery and vein for 90 min. Experimental protocols were divided into four groups: sham operation group, no luminal injection (control) group, luminal injection of 5% glucose saline (GS) solution group, and luminal injection of hydrogen-rich GS (HRGS) group. Two milliliters of experimental solution was locally injected into the lumen of the intestine before declamping of vessels. Oxidative stress markers, proinflammatory cytokines, apoptosis in the crypt cells, and morphologic changes of the intestine were assessed.

RESULTS: The production of malondialdehyde and 8-hydroxydeoxyguanosine, as oxidative stress markers, were markedly suppressed in HRGS group. The level of proinflammatory cytokines, such as inducible nitric oxide synthase and interleukin-6, was significantly inhibited in HRGS group. Crypt apoptosis was also significantly suppressed in HRGS group. Histopathologically, integrity of villus in intestine was maintained in HRGS group in comparison to the other groups.

CONCLUSION: Luminal injection of hydrogen-rich solution can reduce oxidative stress and consequently ameliorate ischemic-reperfusion injury. Hydrogen-containing solution can be a novel and promising luminal preservation material in SBT.

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