氢气抗肿瘤研究进展 | Progress in Hydrogen Anti-tumor Research

Author: Gaoxin Zhou School of Biomedical Engineering and Information, Nanjing Medical University, Nanjing 211166, Jiangsu, China;
作者：周高新南京医科大学生物医学工程与信息学院，江苏南京 211166；

summary 概括

Hydrogen is a new type of gas signal molecule with selective antioxidant and anti-inflammatory effects. The earliest research on hydrogen in the medical field was for anti-tumor use, and subsequently it has shown therapeutic potential for more than 100 diseases in multiple fields, including stroke. The high incidence and mortality of tumors have caused a heavy medical burden. In the short term, there is a lack of effective means to cure tumors. How to prevent tumors and prolong the survival of tumor patients is the primary goal, and hydrogen is expected to play a role in this field. This article reviews the research progress of hydrogen anti-tumor, mainly introducing the anti-cancer mechanism of hydrogen, synergistic and toxic effects, combination therapy, clinical research and hydrogen administration methods.
氢气是一种新型气体信号分子，具有选择性抗氧化和抗炎作用。氢气在医学领域的研究最早是用于抗肿瘤，随后已在多个领域显示出对100多种疾病的治疗潜力，包括中风。肿瘤的高发病率和死亡率造成了沉重的医疗负担，短期内缺乏有效的治愈肿瘤的手段。如何预防肿瘤、延长肿瘤患者的生存期是首要目标，氢气有望在此领域发挥作用。本文就氢气抗肿瘤的研究进展作一综述，主要介绍氢气的抗癌机制、协同毒性作用、联合治疗、临床研究及氢气给药方法等。

Keywords 关键词

#hydrogen ; #hydrogen medicine ; #hydrogen therapy ; #tumor ; #antioxidant
#氢气 ; #氢医学 ; #氢疗法 ; #肿瘤 ; #抗氧化

• 1 Introduction 1 简介

• Hydrogen is the smallest gas molecule. As a clean energy source, it is known for its high calorific value and non-polluting combustion products. In recent years, hydrogen has gradually become a star molecule in the medical field. In particular, with the inclusion of hydrogen-oxygen therapy in the treatment of COVID-19, the clinical application of hydrogen has reached a new level [ 1-3 ] . Hydrogen has weak reducing properties and is considered to be a selective antioxidant that can capture strong oxidizing reactive oxygen species such as hydroxyl radicals (·OH) and peroxynitrite ions (ONOO- ) . Unlike general gas signal molecules (such as NO, CO, H2S, etc. ) [ 4-6 ] , hydrogen has high biosafety and has great advantages over general antioxidants. No adverse reactions to hydrogen have been found in clinical studies. Hydrogen was first used as a diving auxiliary gas in the medical field. The long-term use of high-pressure hydrogen has not been found to have any safety issues for the human body, which also indirectly proves the safety of hydrogen in the body. In fact, hydrogen has been approved for use as a food additive [ 7 ] .
  氢气是最小的气体分子，作为清洁能源，以热值高、燃烧产物无污染著称。近年来，氢气逐渐成为医学领域的明星分子，尤其是随着氢氧疗法被纳入新冠肺炎治疗，氢气的临床应用迈上了一个新台阶 [ 1-3 ] 。 氢气具有弱还原性，被认为是一种选择性抗氧化剂，可以捕获羟基自由基（·OH）和过氧亚硝酸根离子（ONOO- ） 等强氧化性活性氧 。 与一般气体信号分子（如 NO、CO、H2S 等 ） [ 4-6 ]不同，氢气生物安全性高，比一般抗氧化剂有很大优势。临床研究中未发现氢气的不良反应。 氢气最早在医学领域作为潜水辅助气体使用。 长期使用高压氢气，目前还未发现对人体有任何安全问题，这也间接证明了氢气对人体的安全性。事实上，氢气已被批准用作食品添加剂 [ 7 ] 。

• In 1975, Dole et al. [ 8 ] from Baylor University in the United States first reported the medical effects of hydrogen in Science, and found that hydrogen had an inhibitory effect on mouse tumor growth. Continuous breathing of high-pressure hydrogen (8 atmospheres, 97.5% hydrogen, 2.5% oxygen) for 14 days can significantly reduce animal skin tumors or leukemia. This harsh experimental condition was difficult to replicate until 1978 when Roberts et al. [ 9 ] conducted similar studies on mouse models of five solid tumors and one leukemia, but did not obtain similar therapeutic effects as in the study by Dole et al. [ 8 ] . Because the mouse tumor models and modeling methods in the two groups of studies were different, and there was no evidence of the physiological effects of hydrogen, related research stagnated. In 2007, Ohsawa et al. [ 10 ] from Japan Medical University reported in Nature Medicine that inhalation of 2% hydrogen has a therapeutic effect on cerebral ischemia-reperfusion injury. Using electron spin resonance, it was found that its therapeutic effect is related to the removal of hydroxyl radicals (·OH) by hydrogen, and it was confirmed that hydrogen has a selective antioxidant effect, which can specifically remove harmful strong oxidative reactive oxygen species (ROS) such as ·OH and ONOO-, but cannot remove superoxide radicals (O2- · ) , nitric oxide radicals (NO·) and hydrogen peroxide (H2O2 ) that are required to maintain normal physiological activities. The elucidation of the selective antioxidant theory has laid the foundation for the development of hydrogen medicine. In addition to the treatment of ischemia-reperfusion injury, a large number of studies have shown that hydrogen has potential therapeutic effects on more than 100 diseases, such as inflammation, metabolic diseases and neurodegenerative diseases. There are also a lot of studies on the use of hydrogen in the treatment of tumors [ 11-12 ] .
  1975 年， 美国贝勒大学的 Dole 等 [ 8 ] 在《Science》上首次报道了氢气的医学作用，发现氢气对小鼠肿瘤生长有抑制作用。连续呼吸高压氢气（8 个大气压，97.5%氢气、2.5%氧气）14 天，可明显缩小动物皮肤肿瘤或白血病。这种恶劣的实验条件难以复制，直到 1978 年 Roberts 等 [ 9 ] 在患有 5 种实体瘤和 1 种白血病的小鼠模型上进行了类似研究，但没有获得与 Dole 等 [ 8 ] 研究类似的治疗效果 。由于两组研究中小鼠肿瘤模型和建模方法不同，且尚无氢气生理作用的证据，相关研究陷入停滞。2007 年，Ohsawa 等[10]在《Science》上发表了题为《氢气对小鼠肿瘤生长具有抑制作用》的文章。 [ 10 ] 日本医科大学的研究人员在《Nature Medicine》杂志上报道，吸入 2%氢气对脑缺血再灌注损伤有治疗作用。利用电子自旋共振研究发现，其治疗作用与氢气清除羟基自由基（·OH）有关，并证实氢气具有选择性抗氧化作用，可以特异性地清除有害的强氧化性活性氧（ROS）如·OH、ONOO-，但不能清除维持正常生理活动所必需的超氧自由基（O2- · ） 、一氧化氮自由基（NO·）和过氧化氢（H2O2）。选择性抗氧化理论的阐明为氢气医学的发展奠定了基础。除了治疗缺血再灌注损伤外，大量研究表明氢气对炎症、代谢性疾病、神经退行性疾病等 100 多种疾病有潜在的治疗作用。利用氢气治疗肿瘤的研究也有很多[11-12]。

• 2. Anti-tumor effects and mechanisms of hydrogen
  2. 氢气的抗肿瘤作用及机制

• Oxidative stress is closely related to the occurrence and development of malignant tumors. ROS damages DNA, proteins and cell membrane lipids, and is the cause of cell carcinogenesis. Based on its selective antioxidant effect, hydrogen has a significant effect in preventing tumor occurrence. Animal models of liver cancer have shown that hydrogen molecules can significantly reduce the incidence of liver cancer and reduce tumor volume, accompanied by a decrease in oxidative stress indicators [ 13 ] . Hydrogen-rich water can also reduce the incidence of renal cell carcinoma in Wistar rats induced by ferric nitrilotriacetate (Fe-NTA) and inhibit the growth of rat tumors [ 14 ] . Hydrogen-rich water not only inhibits the inflammatory response of kidney tissue and the aggregation of macrophages, but also inhibits the expression of vascular endothelial growth factor (VEGF), the phosphorylation level of signal transduction and transcription activator 3 (STAT3) and the expression of proliferating cell nuclear antigen (PCNA). Frajese et al. from the University of Rome [ 15 ] found that hydrogen-rich electrolyzed water treatment can induce apoptosis of breast cancer cells (human MCF-7, MDA-MB-453 cells and mouse TUBO cells), reduce the expression of human epidermal growth factor receptor-2 (ErbB2 / neu), and disrupt the phosphorylation of extracellular signal-regulated kinase (ERK1/2) and protein kinase B (AKT). The effect of hydrogen-rich electrolyzed water is not affected by the status of p53 tumor suppressor gene, estrogen receptor (ER) and progesterone receptor (PR). Hydrogen selectively reacts with ROS, reducing chromosomal damage to cells, and may also inhibit the abnormal activation of tumor-related signaling pathways, enabling the ataxia telangiectasia mutant gene to repair chromosomal damage in a timely manner, thereby inhibiting the occurrence and development of tumors [ 16 ] .
  氧化应激与恶性肿瘤的发生发展密切相关，ROS 损伤 DNA、蛋白质和细胞膜脂质，是细胞致癌的原因。基于氢气的选择性抗氧化作用，氢气在预防肿瘤发生方面有显著的效果。肝癌动物模型显示，氢分子能显著降低肝癌发病率、缩小肿瘤体积，同时伴随氧化应激指标的下降 [ 13 ] 。富氢水还能降低三乙酸铁（Fe-NTA）诱发的 Wistar 大鼠肾细胞癌的发病率，抑制大鼠肿瘤的生长 [ 14 ] 。富氢水不仅能抑制肾脏组织的炎症反应和巨噬细胞的聚集，还能抑制血管内皮生长因子（VEGF）的表达、信号转导和转录激活因子 3（STAT3）的磷酸化水平和增殖细胞核抗原（PCNA）的表达。Frajese 等。 来自罗马大学的研究人员 [ 15 ] 研究发现，富氢电解水处理可以诱导乳腺癌细胞(人 MCF-7、MDA-MB-453 细胞和小鼠 TUBO 细胞)凋亡，降低人表皮生长因子受体-2(ErbB2 neu)的表达 ，破坏细胞外信号调节激酶(ERK1/2)和蛋白激酶 B(AKT)的磷酸化。富氢电解水的效果不受 p53 肿瘤抑制基因、雌激素受体(ER)和孕激素受体(PR)状态的影响。氢气选择性地与 ROS 发生反应，减少染色体对细胞的损伤，也可能抑制肿瘤相关信号通路的异常激活，使毛细血管扩张性共济失调突变基因及时修复染色体损伤，从而抑制肿瘤的发生和发展 [ 16 ] 。

• Abnormal metabolism of tumor tissue causes tumor cells to be in a state of oxidative stress. High levels of ROS can promote tumor proliferation, migration and invasion. Therefore, destroying the redox balance of tumor tissue has become a strategy to inhibit tumor growth and metastasis. Japanese scholars Nishikawa et al. [ 17 ] conducted a series of studies using hydrogen-rich water containing platinum nanoparticles (particle size of about 2 nm) and found that platinum-containing hydrogen water can inhibit chemically induced malignant transformation of mouse embryonic fibroblasts; at the same time, this platinum-containing hydrogen water has a killing effect on human tongue cancer cell HSC-4, human esophageal squamous cell carcinoma cell KYSE70, human promyelocytic leukemia cell HL60 and human gastric adenocarcinoma-derived NUGC-4 cells [ 18-20 ] , but has almost no effect on normal cells, which may be related to the specific internalization of colloidal platinum by tumor cells. In vitro experiments found that colloidal platinum reduced the redox potential of hydrogen water and enhanced its activity in capturing DPPH free radicals. However, the level of ROS in cells increased instead of decreasing, indicating that hydrogen water activated by colloidal platinum can induce redox imbalance in tumor cells [ 18 , 21 ] .
  肿瘤组织代谢异常导致肿瘤细胞处于氧化应激状态，高水平的 ROS 可促进肿瘤的增殖、迁移和侵袭，因此破坏肿瘤组织的氧化还原平衡成为抑制肿瘤生长和转移的一种策略。日本学者西川等 [ 17 ] 利用含铂纳米粒子（粒径约 2nm）的富氢水进行了一系列研究，发现含铂氢水能抑制化学诱导的小鼠胚胎成纤维细胞恶性转化；同时，这种含铂氢水对人舌癌细胞 HSC-4、人食管鳞状细胞癌细胞 KYSE70、人早幼粒细胞白血病细胞 HL60 及人胃腺癌来源的 NUGC-4 细胞均有杀伤作用[ ] ， 但对正常细胞几乎没有影响，这可能与肿瘤细胞对胶体铂的特异性内化，体外实验发现胶体铂降低了氢水的氧化还原电位，增强了其捕获 DPPH 自由基的活性。 但细胞内 ROS 水平不降反升，表明胶体铂活化的氢水可以诱导肿瘤细胞内氧化还原失衡 [ 18,21 ] 。

• Regarding the treatment of lung cancer with hydrogen, Zhang Yu et al. [ 22 ] from the Third Hospital of Hebei Medical University conducted a large number of studies. They investigated the effect of hydrogen on non-small cell lung cancer and found that in vitro hydrogen culture (concentrations ranging from 20% to 60%) can promote apoptosis of lung cancer A549 and NCI-H1975 cells, and reduce the expression of X-link inhibitor of apoptosi protein (XIAP) and baculovirus IAP repeat sequence 3 (BIRC3). High concentrations (60%) of hydrogen can also cause an increase in the expression of p21, caspase7, and caspase9 proteins in lung cancer A549 cell lines and A549 cell metastatic tumor tissues, as well as a decrease in the expression of cyclin dependent kinase 4 (CDK4) [ 23 ] . Wang et al. [ 24 ] also found that the anti-non-small cell lung cancer effect of hydrogen is related to chromosome condensation. Hydrogen intervention can inhibit the expression of chromosome structure maintenance protein 3 (SMC3) gene and protein in A549 and H1975 cells, thereby changing the cell cycle. Hydrogen can also inhibit the expression of Ki-67 antigen, cyclooxygenase-2 (COX-2), and VEGF in lung cancer tissues in mice [ 25 ] . Ye et al. [ 26 ] found that hydrogen-containing electrolyzed water can downregulate the transcription and protein expression of VEGF gene in A549 cells by inhibiting the activation of extracellular signal-regulated kinase (ERK). In addition, hydrogen can promote lung cancer cell apoptosis or autophagy by inhibiting the activation of STAT3/Bcl2 pathway, and inhibiting autophagy can further enhance the apoptosis of lung cancer cells [ 27 ] . Hydrogen therapy can also inhibit the progression of lung cancer by inhibiting the activation of the CD47/CDC42 pathway, and is expected to become an effective treatment for patients with tumors with high CD47 expression [ 28 ] . The above studies provide a basis for hydrogen treatment of lung cancer. The efficacy of hydrogen on lung cancer needs further verification in vivo. There are also individual cases in clinical practice showing that hydrogen inhalation is beneficial to lung cancer patients, but there is still a lack of evidence-based medical evidence [ 29 ] .
  关于氢气治疗肺癌，河北医科大学第三医院张宇等 [ 22 ] 进行了大量研究，研究了氢气对非小细胞肺癌的影响，发现体外氢气培养（浓度为 20%～60%）可促进肺癌 A549 和 NCI-H1975 细胞凋亡，降低 X-link 凋亡抑制蛋白（XIAP）和杆状病毒 IAP 重复序列 3（BIRC3）的表达。高浓度（60%）氢气还能引起肺癌 A549 细胞株和 A549 细胞转移瘤组织中 p21、caspase7、caspase9 蛋白表达增加，细胞周期依赖性蛋白激酶 4（CDK4）表达降低[23]。王等[24]研究发现，氢气在体外培养（浓度为 20%～60%）可诱导肺癌 A549 和 NCI-H1975 细胞凋亡，降低 XIAP 和 BIRC3 的表达。高浓度（ 60 % ） 氢气还可引起肺癌 A549 细胞株和 A549 细胞转移瘤组织中 p21、caspase7、caspase9 蛋白表达增加，细胞周期依赖性蛋白激酶 4（CDK4）表达降低 。 [ 24 ] 还研究发现氢气的抗非小细胞肺癌作用与染色体凝聚有关，氢气干预可以抑制 A549 和 H1975 细胞中染色体结构维持蛋白 3（SMC3）基因和蛋白的表达，从而改变细胞周期。 氢气还能抑制小鼠肺癌组织中 Ki-67 抗原、环氧合酶-2（COX-2）和 VEGF 的表达 [ 25 ] 。叶等人 [ 26 ] 研究发现，含氢电解水可通过抑制细胞外信号调节激酶（ERK）的活化，下调 A549 细胞中 VEGF 基因的转录和蛋白表达。此外，氢气还能通过抑制 STAT3/Bcl2 通路的活化，促进肺癌细胞凋亡或自噬，而抑制自噬可进一步增强肺癌细胞的凋亡 [ 27 ] 。氢气疗法还能通过抑制 CD47/CDC42 通路的活化，抑制肺癌的进展，有望成为 CD47 高表达肿瘤患者的有效治疗方法 [ 28 ] 。以上研究为氢气治疗肺癌提供了依据。 氢气对肺癌的疗效还有待进一步在体内验证，临床上也有个别病例显示氢气吸入对肺癌患者有益，但目前还缺乏循证医学证据 [ 29 ] 。

• Zhou Xiao et al. [ 30 ] used hydrogen water to culture primary colon cancer cells from 15 patients in vitro. The results showed that hydrogen had an inhibitory effect on the growth of primary human colon adenocarcinoma cells, and the patient's gender, age, tumor location and Dukes stage had no significant effect on whether the primary cancer cells were inhibited by hydrogen. Zhang Yao et al. [ 31 ] found that drinking hydrogen-rich water could inhibit the growth of tumors in mice bearing human colorectal cancer SW480 cells. On the 28th day, the tumor volume in the hydrogen intervention group was significantly smaller than that in the control group. Liu et al. [ 32 ] conducted an in vivo study and constructed a C6 orthotopic glioma model and a U87 subcutaneous tumor model. They found that mice inhaling 67% hydrogen twice a day (1 hour each time) could inhibit the growth of gliomas and prolong their lifespan. Hydrogen can downregulate the expression of CD133 and Nestin (cell stemness markers), Ki67 (cell proliferation marker) and CD34 (angiogenesis marker), while upregulating the expression of GFAP (cell differentiation marker). Hydrogen also inhibited the in vitro spheroidization ability, migration, invasion and cloning ability of glioma cells, indicating that hydrogen can act by reducing the stemness of glioma cells. Studies have also found that hydrogen culture can reduce the stemness of human liver cancer cells Huh7 and downregulate the expression of cell intermediate filament vimentin, providing evidence for the use of hydrogen in the prevention and treatment of liver cancer [ 33 ] . Li Jiawei et al. [ 34 ] found that hydrogen can promote macrophage polarization to M1, inhibit its polarization to M2, and ultimately promote apoptosis of breast cancer cells. Studies on endometrial cancer found that hydrogen molecules induce tumor cell pyroptosis through the ROS/NLRP3/caspase⁃1/GSDMD pathway. After hydrogen-rich water culture, the levels of ROS and mitochondrial ROS (mtROS) in human endometrial cancer cells (AN3CA, HEC1A and Ishikawa cells) were increased [ 35 ] .
  周晓等 [ 30 ] 用氢水体外培养 15 名患者原发性结肠癌细胞，结果显示氢气对人原发性结肠腺癌细胞的生长有抑制作用，且患者的性别、年龄、肿瘤部位和 Dukes 分期对原发性癌细胞是否被氢气抑制无明显影响。张耀等 [ 31 ] 研究发现，饮用富氢水可抑制携带人结直肠癌 SW480 细胞的小鼠肿瘤的生长，第 28 天，氢气干预组肿瘤体积明显小于对照组。刘等 [ 32 ] 进行了体内研究，构建了 C6 原位胶质瘤模型和 U87 皮下肿瘤模型，发现小鼠每天吸入 2 次 67%氢气（每次 1 小时），可抑制胶质瘤的生长，延长其寿命。 氢气可以下调细胞干性标志物 CD133 和 Nestin、细胞增殖标志物 Ki67 以及血管生成标志物 CD34 的表达，而上调细胞分化标志物 GFAP 的表达，同时抑制胶质瘤细胞体外球化能力、迁移、侵袭和克隆能力，表明氢气可以通过降低胶质瘤细胞的干性而起作用。有研究发现，氢气培养可以降低人肝癌细胞 Huh7 的干性，下调细胞中间丝波形蛋白的表达，为氢气用于肝癌的防治提供了证据 [ 33 ] 。李嘉伟等 [ 34 ] 研究发现，氢气可以促进巨噬细胞向 M1 极化，抑制其向 M2 极化，最终促进乳腺癌细胞凋亡。对子宫内膜癌的研究发现，氢分子通过 ROS/NLRP3/caspase-1/GSDMD 通路诱导肿瘤细胞焦亡。 经过富氢水培养后，人子宫内膜癌细胞（AN3CA、HEC1A 和 Ishikawa 细胞）中的 ROS 和线粒体 ROS（mtROS）水平升高 [ 35 ] 。

• The most direct evidence of hydrogen's involvement in physiological activities is its antioxidant activity. Currently, the antioxidant theory alone cannot explain the anti-tumor mechanism of hydrogen. In contrast, the study of the effects of traditional antioxidants on tumors has a long history, but has not achieved satisfactory clinical results
  氢气参与生理活动最直接的证据就是其抗氧化作用，目前单纯的抗氧化理论还不能解释氢气的抗肿瘤机制，而传统抗氧化剂对肿瘤作用的研究历史悠久，但尚未取得令人满意的临床效果[ 36 ] . For example, long-term studies on the anti-cancer effects of vitamin C have found that its mechanism is that vitamin C itself promotes oxidative stress levels after being oxidized, and the generation of hydrogen peroxide is the key to vitamin C's anti-cancer effect
  例如，对维生素 C 抗癌作用的长期研究发现，其机制是维生素 C 本身被氧化后会促进氧化应激水平的提高，而过氧化氢的生成正是维生素 C 发挥抗癌作用的关键[ 37-38 ] . Therefore, the antioxidant mechanism of hydrogen in treating tumors is not perfect, and there may be other unknown mechanisms for hydrogen's anti-cancer effect. Hydrogen medical research has shown that hydrogen participates in the regulation of signal pathways and can affect protein structure and enzyme activity, which may provide new ideas for hydrogen anti-tumor research. For example, in vitro protein structure studies have found that hydrogen molecules can regulate the mechanical properties of proteins and enhance the activity of pepsin
  氢气的抗癌作用。氢气医学研究表明，氢气参与信号通路的调控，能够影响蛋白质结构和酶活性，这或许为氢气抗肿瘤研究提供新思路。例如，体外蛋白质结构研究发现，氢分子能够调节蛋白质的力学性质，增强胃蛋白酶的活性[ 39 ] . In addition to exerting direct antioxidant effects, hydrogen can also exert its effects by regulating antioxidant enzymes. For example, hydrogen can inhibit the phosphorylation of human membrane-associated protein tyrosine kinase (Lyn) associated with the immunoglobulin Fc segment receptor FcεRI (IgE high affinity receptor) and downregulate the activity of NADPH oxidase in mast cells, thereby reducing the generation of H2O2
  氢气除了发挥直接的抗氧化作用外，还可以通过调节抗氧化酶发挥作用。例如氢气可以抑制与免疫球蛋白 Fc 段受体 FcεRI（IgE 高亲和力受体）相关的人体膜相关蛋白酪氨酸激酶（Lyn）的磷酸化，下调肥大细胞中 NADPH 氧化酶的活性，从而减少 H2O2 的生成, rather than directly capturing ROS [ 40 ] . Ostojic et al. [ 41 ] reported that hydrogen may regulate mitochondrial energy metabolism through at least four pathways, among which ghrelin through its receptor GHS-R1α is the most important pathway. Hydrogen can inhibit the generation of NO signaling molecules in RAW264 macrophages by regulating the activity of inducible nitric oxide synthase (iNOS) [ 42 ] . Hydrogen can also mediate the phosphorylation and degradation of β-catenin, thereby inhibiting the activation of the Wnt/β-catenin signaling pathway [ 43 ] . The latest research found that hydrogen can act on thioredoxin 1, reducing the expression of tissue factor and matrix metalloproteinase (MMP)-9 [ 44 ] . Hydrogen can also reduce the expression of MMP-3 and MMP-13 [ 45 ] . Metalloproteinases can degrade the extracellular matrix and play an important role in mediating tumor angiogenesis, metastasis and invasion. Recently, Ma Xuemei et al. [ 46 ] found that hydrogen can increase the activity of biological enzymes including mitochondrial complex I, acetylcholinesterase and horseradish peroxidase, suggesting that the physiological effect of hydrogen may be a multi-target enzymatic reaction process. In addition, hydrogen can also fight tumors by regulating the human immune state. In a clinical study conducted in Japan from 2014 to 2017, 55 patients with stage IV colon cancer were included. All patients received XELOX (CapeOX) chemotherapy and used a home hydrogen inhalation machine to inhale hydrogen for 3 hours every day. The correlation between the expression of PD-1 in CD8 + T cells in the patients' blood and progression-free survival and overall survival was analyzed. It was found that hydrogen inhalation can reduce the proportion of PD-1 + /CD8 + double-positive T cells in colon cancer patients and improve the prognosis of patients [ 47 ] . It is speculated that this may be because hydrogen activates the signal molecule PGC-1α, which in turn upregulates the respiration of mitochondria in T cells, ultimately rescuing exhausted CD8 + T cells and exerting its effect. At the same time, the improvement in the overall survival rate of patients is related to the increase in coenzyme Q10 levels, which can allow more hydrogen molecules to enter the mitochondria to exert their effects [ 48 ] . The above studies suggest that hydrogen can not only exert its effects through direct antioxidant effects. In addition to selective antioxidant effects, hydrogen may also act as a gas signal molecule similar to "NO". The anti-tumor mechanism of hydrogen needs further in-depth research.

• 3 Hydrogen is used to enhance the efficacy and reduce the toxicity of radiotherapy and chemotherapy
  3 氢气用于放化疗增效减毒

• At present, surgery, chemotherapy and radiotherapy are still the main means of cancer treatment, but the serious adverse reactions caused by traditional radiotherapy and chemotherapy have not been solved. Based on the antioxidant strategy, molecular hydrogen has the effect of enhancing the efficacy and reducing the toxicity of tumor radiotherapy and chemotherapy. Qian et al.
  目前手术、化疗、放疗仍是癌症治疗的主要手段，但传统放化疗引起的严重不良反应仍未得到解决。基于抗氧化策略，分子氢具有增强肿瘤放化疗疗效、降低毒性的作用。钱等[ 49 ] proposed that hydrogen is an effective and safe radiation protector that can significantly inhibit ionizing radiation-induced apoptosis of HIEC intestinal epithelial cells, increase the endogenous antioxidant level of AHH-1 human peripheral blood B lymphocytes, and alleviate the depletion of leukocytes and platelets caused by radiation
  提出氢气是一种有效、安全的辐射防护剂，能显著抑制电离辐射诱导的 HIEC 肠上皮细胞凋亡，提高 AHH-1 人外周血 B 淋巴细胞内源性抗氧化水平，缓解辐射引起的白细胞和血小板的耗竭[ 50 ] . In addition to protecting cells cultured in vitro from ionizing radiation damage, in vivo studies have also found that intraperitoneal injection of hydrogen-rich saline into male BALB/c mice can inhibit the occurrence and growth of radiation-induced thymic lymphoma
  除了可以保护体外培养的细胞免受电离辐射损伤外，体内研究还发现，给雄性 BALB/c 小鼠腹腔注射富氢盐水，可以抑制放射性胸腺淋巴瘤的发生和生长[ 51-53 ] . Zhang et al. 等[ 54 ] found that hydrogen-rich solution also has a good protective effect on radiation damage to normal human immortalized epidermal keratinocytes HaCaT and rat head and neck skin, and its mechanism is related to oxidative damage of biological molecules and the scavenging of free radicals. Hydrogen can also alleviate radiation-related liver damage during radiotherapy, and has a protective effect on human liver LO2 cells and mouse livers
  研究发现富氢溶液对正常人永生化表皮角质形成细胞 HaCaT 及大鼠头颈部皮肤的放射损伤也有良好的防护作用，其机制与生物分子的氧化损伤和自由基的清除有关。氢气在放疗过程中还能减轻放射性肝损伤，对人肝 LO2 细胞及小鼠肝脏有保护作用[ 55 ] . It has no inhibitory effect on the growth of human liver cancer HepG2 cells, but can inhibit fat accumulation in HepG2 cells by downregulating the expression of CD36 protein
  对人肝癌 HepG2 细胞生长无抑制作用，但能通过下调 CD36 蛋白的表达，抑制 HepG2 细胞内脂肪蓄积[ 56 ] . Intragastric administration of hydrogen-rich water can reduce bone marrow suppression in C57BL/6 mice irradiated with 4Gy X-rays
  . 富氢水灌胃可减轻 4Gy X 射线照射的 C57BL/6 小鼠的骨髓抑制[ 57 ] . Terasaki et al. . Terasaki 等人[ 58 ] proved through cell and mouse experiments that drinking hydrogen-rich water combined with 3% hydrogen inhalation can not only counteract acute radiation damage, but also has an effect on chronic pulmonary fibrosis. A retrospective study on hydrogen intervention with intensity-modulated radiotherapy was conducted in Tokyo, Japan. A total of 26 patients with advanced cancer were included between 2015 and 2016. It was found that hydrogen inhalation (0.5 h per day) can significantly improve bone marrow damage caused by intensity-modulated radiotherapy and has no effect on the efficacy of radiotherapy
  通过细胞和小鼠实验证明，饮用富氢水结合3%氢气吸入不仅可以对抗急性放射损伤，对慢性肺纤维化也有一定作用。日本东京进行了一项关于氢气干预​​调强放疗的回顾性研究，2015年至2016年共纳入26例晚期癌症患者，结果发现氢气吸入（每天0.5小时）可以明显改善调强放疗引起的骨髓损伤，且对放疗疗效无影响[ 59 ] . Kang et al. . 康等人[ 60 ] from the University of Pittsburgh conducted a two-arm randomized controlled clinical trial that recruited 49 liver cancer patients in 2006. After radiotherapy, the patients were given hydrogen-rich water for 6 weeks. The patients were required to take 1.5-2.0L of hydrogen water several times a day. The results showed that the quality of life scores of the patients in the hydrogen water intervention group after radiotherapy were significantly improved. Radiotherapy can kill tumor cells through high-energy rays, while causing damage to normal cells in the tissues around the lesions. The antioxidant mechanism of hydrogen is clear, and it can play a good role in enhancing the efficacy and reducing toxicity when used as an adjuvant for radiotherapy. At the same time, hydrogen has the advantages of high safety and ease of use compared with other adjuvant treatment methods.
  美国匹兹堡大学的研究人员于 2006 年进行了一项双臂随机对照临床试验，招募了 49 名肝癌患者，在放疗后给予患者富氢水 6 周，要求患者每天多次饮用 1.5-2.0L 氢水。结果显示，放疗后氢水干预组患者的生活质量评分明显改善。放疗可以通过高能射线杀死肿瘤细胞，同时对病灶周围组织中的正常细胞造成损伤。氢气的抗氧化机制明确，作为放疗的辅助手段可以起到很好的增效减毒作用。同时，氢气相较于其他辅助治疗方法具有安全性高、使用方便等优势。

• In terms of tumor chemotherapy, Nakashima et al. [ 61 ] investigated the effect of inhaling 1% hydrogen (the entire experimental period) after cisplatin treatment on the adverse reactions of mice transplanted with S-180 mouse sarcoma cells. They found that inhaling hydrogen could improve the survival rate of mice, and the weight loss and neurotoxicity caused by cisplatin were also alleviated. Control experiments have shown that the intake of hydrogen water can achieve similar effects. Hydrogen water also has a mitigating effect on the cardiac toxicity caused by doxorubicin. Rats receiving doxorubicin chemotherapy were intraperitoneally injected with 10 mL/kg of hydrogen-saturated saline every day. After one month, the levels of oxidative stress markers in the blood and the autophagic activity of myocardial cells were significantly reduced [ 62 ] . Molecular hydrogen inhibits the NF-κB/IL-6 inflammatory pathway, enhances the expression of activated caspase-8 and the ratio of Bcl-2/Bax, thereby alleviating cell apoptosis in heart and liver tissues [ 63 ] . Fan et al. [ 64 ] prepared MgB2 nanosheets into oral preparations for combined treatment of doxorubicin chemotherapy and hydrogen for in situ gastric cancer in mice, which effectively reduced the cardiac toxicity of doxorubicin. Molecular hydrogen can not only promote apoptosis of colon cancer cells, but also significantly enhance the anti-tumor effect of 5-fluorouracil when used in combination with hydrogen-rich water. The higher the concentration of hydrogen-rich water, the more obvious the synergistic effect. This effect was verified in a Colon 26 cell tumor-bearing mouse model
  将氢分子水与阿霉素化疗联合用于治疗小鼠原位胃癌，有效降低了阿霉素对心脏的毒性。氢分子水不仅能促进结肠癌细胞凋亡，与富氢水联合使用还能显著增强5-氟尿嘧啶的抗肿瘤作用，富氢水浓度越高，协同作用越明显。此效果在结肠26细胞荷瘤小鼠模型中得到验证[ 65 ] . Meng Xiaoyin et al.
  .孟小银等.[ 66-67 ] found that hydrogen-saturated saline can effectively prevent cyclophosphamide chemotherapy-induced ovarian damage in rats. A randomized controlled single - blind trial involving 144 patients with colon cancer conducted by Taishan Hospital from 2010 to 2016 showed that there was no significant change in liver function before and after treatment in patients receiving mFOLFOX6 standard chemotherapy in the hydrogen-rich water group, while the levels of alanine aminotransferase, aspartate aminotransferase and indirect bilirubin in the placebo control group were significantly increased, proving that molecular hydrogen has a protective effect on the liver during chemotherapy
  泰山医院 2010 年至 2016 年对 144 例结肠癌患者进行的盲法试验结果显示，富氢水组接受 mFOLFOX6 标准化疗的患者治疗前后肝功能无明显变化，而安慰剂对照组的丙氨酸氨基转移酶、天冬氨酸氨基转移酶和间接胆红素水平均明显升高，证明了分子氢在化疗过程中对肝脏具有保护作用[ 68 ] . However, patients need to drink 1L (4×250mL) of hydrogen-rich water every day from 1 day before treatment to 4 days after treatment. A systematic retrospective analysis of 80 patients with malignant tumors from 2013 to 2014 by Yang Qingxi et al.
  但患者需在治疗前 1 天至治疗后 4 天，每天饮用 1L(4×250mL)富氢水。杨庆熙等对 2013 年至 2014 年 80 例恶性肿瘤患者进行了系统回顾性分析。[ 69 ] showed that drinking hydrogen-rich water on the basis of conventional chemotherapy can relieve bone marrow suppression in patients after chemotherapy, and has a protective effect on heart, gastrointestinal tract, liver and kidney function, and improves physical decline caused by chemotherapy. A study by Yao et al.
  研究表明，在常规化疗基础上饮用富氢水，可减轻化疗后患者的骨髓抑制，并对心脏、胃肠道、肝肾功能有保护作用，改善化疗引起的体力衰退。姚期智等的研究[ 70 ] found that hydrogen-rich electrolyzed water can enhance the liver's excretion of exogenous toxins by increasing the expression levels of P-glycoprotein and multidrug resistance-associated protein-2, thereby achieving a detoxification effect. Tyrosine kinase inhibitor targeted drugs have significant efficacy in the treatment of non-small cell lung cancer. Such drugs can also cause adverse reactions when taken orally in large quantities. For example, gefitinib may cause severe acute interstitial pneumonia. Terasaki et al.
  研究发现富氢电解水可通过增加 P-糖蛋白和多药耐药相关蛋白-2 的表达水平，增强肝脏对外源性毒素的排泄，从而达到解毒的效果。酪氨酸激酶抑制剂类靶向药物在非小细胞肺癌治疗中疗效显著，该类药物大剂量口服也会引起不良反应，如吉非替尼可能引起重症急性间质性肺炎。Terasaki 等[ 71 ] found through mouse experiments that hydrogen-rich water can alleviate the adverse reactions of gefitinib. Chen et al.
  陈等通过小鼠实验发现富氢水可以减轻吉非替尼的不良反应。[ 72 ] found in clinical studies that hydrogen can effectively reduce adverse drug reactions in patients with advanced non-small cell lung cancer. In terms of adjuvant treatment of cancer patients after radiotherapy and chemotherapy, Chen et al.
  临床研究发现氢气能有效减少晚期非小细胞肺癌患者的药物不良反应。在癌症患者放化疗后的辅助治疗方面，陈等[14]研究发现氢气能有效减少晚期非小细胞肺癌患者的药物不良反应。[ 73 ] have accumulated many real-world hydrogen treatment cases, showing that hydrogen is beneficial to improving the quality of life of cancer patients. In terms of surgical treatment, Niu Tongxiang et al.
  已积累了大量的真实氢气治疗案例，显示氢气有利于改善癌症患者的生活质量。在外科治疗方面，牛同祥等[ 74 ] studied 110 patients undergoing thoracoscopic radical resection of lung cancer and found that inhaling hydrogen for 1 hour before surgery can reduce the occurrence of postoperative pulmonary complications. The above basic and clinical research results suggest that hydrogen has a good effect in enhancing the efficacy and reducing the toxicity of tumor radiotherapy and chemotherapy, and has good compatibility with other treatment methods. It should be promoted as a priority in clinical application.
  对110例胸腔镜肺癌根治术患者进行研究发现，术前吸入氢气1小时，可减少术后肺部并发症的发生。以上基础及临床研究结果提示，氢气在增强肿瘤放化疗疗效、降低毒性方面具有良好的作用，且与其他治疗方法有良好的兼容性，应在临床应用中优先推广。

• 4 Hydrogen combined therapy
  4 氢气联合治疗

• Due to the multi-target action mode of hydrogen, the effect of hydrogen therapy may be limited. In addition to safety and availability, the important value of hydrogen in the field of tumor treatment needs to be reflected through combined treatment. For example, the combination of hydrogen and the PI3K inhibitor LY294002 can enhance the inhibition of A549 cells
  由于氢气的多靶点作用方式，氢气治疗的效果可能受到限制。除了安全性和可用性之外，氢气在肿瘤治疗领域的重要价值需要通过联合治疗来体现。例如氢气与 PI3K 抑制剂 LY294002 联合使用，可以增强对 A549 细胞的抑制[ 75 ] . In an in situ gastric cancer animal model, MgB2 nanosheets that can release hydrogen are ingested through feed
  在原位胃癌动物模型中，通过饲料摄入能够释放氢气的 MgB2 纳米片. The hydrogen released in situ in the acidic environment of gastric fluid can directly act on gastric tumor cells, produce a synergistic effect with the chemotherapy drug doxorubicin, enhance the therapeutic effect, and effectively alleviate the adverse cardiac reactions of doxorubicin, truly achieving the effect of increasing efficacy and reducing toxicity
  在胃液酸性环境中原位释放的氢气可直接作用于胃肿瘤细胞，与化疗药物阿霉素产生协同作用，增强治疗效果，并有效缓解阿霉素的心脏不良反应，真正达到增效减毒的效果[ 64 ] . Similarly, under in vitro culture conditions, the hydrogen released by a magnesium micromotor can have a synergistic effect with doxorubicin. This micromotor can produce hydrogen in situ in cells by active movement, which increases the chemotherapy effect of doxorubicin by 2.4 times and effectively kills mouse breast cancer 4T1 cells
  同样，在体外培养条件下，镁微马达释放的氢气可以与阿霉素产生协同作用。这种微马达通过主动运动，在细胞内原位产生氢气，使阿霉素的化疗效果提高 2.4 倍，有效杀伤小鼠乳腺癌 4T1 细胞[ 76 ] . Based on the principle of photocatalytic hydrogen production, using the strong ability of nanomaterials to cross mucosa and penetrate tumor cells, hydrogen can be generated in situ in tumor cells under 660nm laser irradiation. Combining hydrogen therapy with perfusion therapy for bladder cancer can significantly enhance the chemotherapy effect
  基于光催化产氢原理，利用纳米材料强大的穿越黏膜和穿透肿瘤细胞的能力，在 660nm 激光照射下可在肿瘤细胞内原位产生氢气。氢气疗法与膀胱癌灌注治疗相结合，可显著增强化疗效果[ 77 ] . Mechanism studies have found that hydrogen can inhibit mitochondrial function, hinder ATP synthesis, weaken the function of P-glycoprotein efflux pump, make it difficult for the chemotherapy drug gemcitabine to be excreted, and achieve efficient synergistic treatment of bladder cancer. Using the hydrogen storage properties of palladium-based nanomaterials, palladium nanoparticles (PdH0.2) and palladium-based metal organic framework materials (PdHMOF) are used for hydrogen-photothermal therapy of tumors, i.e. hydrogen thermal therapy, which can enhance the anti-cancer effect. In addition, active hydrogen can disrupt the redox balance of tumor cells and destroy the energy metabolism
  。机制研究发现，氢气可抑制线粒体功能，阻碍 ATP 合成，削弱 P-糖蛋白外排泵功能，使化疗药物吉西他滨难以排出，实现膀胱癌的高效协同治疗。利用钯基纳米材料的储氢特性，将钯纳米粒子（PdH0.2）和钯基金属有机骨架材料（PdHMOF）用于肿瘤的氢光热治疗，即氢热疗法，可增强抗癌效果。此外，活性氢能破坏肿瘤细胞的氧化还原平衡，破坏能量代谢of cells [ 78-79 ] . Zhang et al. 张等人[ 80 ] used dopamine-loaded ammonia borane to achieve hyperthermia. The hydrogen released by the latter effectively changed the redox homeostasis of the tumor site, which was beneficial to combating inflammation. Ultimately, while inhibiting tumor growth, combined therapy also achieved inhibition of distal tumors. Recently, using transformation nanomaterials, a hydrogen-mediated cascade amplification multimodal synergistic treatment strategy can use hydrogen generated by near-infrared photolysis of water to cooperate with photodynamic, photothermal and chemodynamic treatment methods to achieve efficient treatment of breast cancer in mice
  采用载多巴胺的氨硼烷实现热疗，后者释放的氢气有效改变了肿瘤部位的氧化还原稳态，有利于对抗炎症，最终在抑制肿瘤生长的同时，联合治疗也实现了对远端肿瘤的抑制。近期，利用转化纳米材料，氢介导的级联放大多模态协同治疗策略，可利用近红外光解水产生的氢气配合光动力、光热和化学动力学治疗方法，实现对小鼠乳腺癌的高效治疗[ 81 ] . Using the same platform to achieve synergy between hydrogen therapy and photodynamic therapy, an amazing therapeutic effect was achieved. Similar results were further verified in a composite system of PCN-224 loaded with nanopalladium
  利用同一平台实现氢疗法与光动力疗法的协同作用，取得了惊人的治疗效果。在 PCN-224 负载纳米钯的复合体系中进一步验证了类似的结果[ 82 ] . Nanopalladium was used as a hydrogen carrier and porphyrin MOF was used as a photosensitizer for PDT treatment. The reason for the synergistic effect of hydrogen therapy and PDT may be that hydrogen stimulation disrupts the redox balance of the tumor, and photodynamic therapy exacerbates this imbalance. Zhao et al.
  .以纳米钯为氢载体、卟啉 MOF 为光敏剂进行 PDT 治疗。氢疗与 PDT 产生协同作用的原因可能是氢刺激破坏了肿瘤的氧化还原平衡，而光动力治疗加剧了这种不平衡。Zhao et al.[ 83 ] also used a Z-shaped nanophotocatalyst to produce hydrogen in situ in tumors, achieving hydrogen-cavity combined therapy. Similarly, Wu et al.
  同样利用 Z 型纳米光催化剂在肿瘤内原位产生氢气，实现了氢腔联合治疗。同样，吴等人[ 84 ] used a radiation hydrogen-producing material to achieve the synergistic effect of hydrogen therapy and radiotherapy. In clinical practice, since hydrogen can improve the immune function of tumor patients, the combination of hydrogen and PD-1 antibodies has been found to improve the clinical effect of immunotherapy and prolong the survival time of patients
  利用放射性产氢物质，实现氢气治疗与放疗的协同作用。在临床上，由于氢气可以提高肿瘤患者的免疫功能，氢气与 PD-1 抗体的联合应用被发现可以提高免疫治疗的临床效果，延长患者的生存时间[ 48 ] . In clinical practice, hydrogen is safe and easy to ingest for the human body. Combining it with existing anti-cancer methods can achieve twice the result with half the effort. At present, hydrogen-combined multifunctional tumor treatment requires the use of nano hydrogen-producing materials, or loading of hydrogen prodrug molecules, or the nanomaterials themselves can release hydrogen under special conditions such as light and acid. The introduction of nanomaterials should be based on the premise of not sacrificing the safety of hydrogen therapy. In the combined treatment of hydrogen and photodynamic therapy, the antioxidant hydrogen and the pro-oxidative photosensitive materials work together to disrupt the redox homeostasis of tumor cells, inhibiting tumor growth and providing a new idea for tumor treatment.
  。在临床上，氢气对人体安全、易摄取，与现有的抗癌手段相结合，可以达到事半功倍的效果。目前，氢气联合多功能肿瘤治疗需要采用纳米产氢材料，或负载氢气前药分子，或纳米材料本身在光、酸等特殊条件下能释放氢气。纳米材料的引入应以不牺牲氢气治疗安全性的前提下进行。在氢气与光动力疗法联合治疗中，具有抗氧化作用的氢气与具有促氧化作用的光敏材料共同作用，破坏肿瘤细胞的氧化还原稳态，抑制肿瘤生长，为肿瘤治疗提供了新思路。

• 5 Methods of hydrogen administration in hydrogen therapy
  5 氢疗法中氢气给药的方法

• In clinical research on hydrogen medicine, the existing methods of hydrogen intake mainly include breathing hydrogen-containing mixed gases or taking hydrogen solutions orally
  在氢医学临床研究中，现有的氢气摄入方式主要有呼吸含氢混合气体或口服氢气溶液[ 85 ] . Hydrogen inhalation requires special equipment and long-term inhalation to be effective. The gas composition and hydrogen ratio are not fixed. There are hydrogen and nitrogen mixed gases. Home hydrogen inhalers produce hydrogen and oxygen mixed gases with a hydrogen-oxygen ratio of 2:1. There are also hydrogen generators that can separate oxygen to produce high-purity hydrogen. In in vivo experiments, the inhalation ratio of hydrogen should not exceed the explosion limit of 4%, otherwise it will be dangerous in actual operation. The saturation concentration of hydrogen-rich water is low (0.8mmol/L), which is not convenient for storage. It must be administered in large doses and is difficult to apply to patients with difficulty swallowing and who are not suitable for large-scale infusion. In addition, the quality standards and storage difficulties of hydrogen-rich water limit its application. At present, there are many methods for preparing hydrogen water, and there is a lack of unified quality standards for hydrogen-rich water used in hydrogen medical research. The main methods for producing hydrogen-rich water are bubbling method and high-pressure dissolution method. The solvents used include pure water, physiological saline, and even organ preservation solution (Celsior)
  。氢气吸入需特殊设备，长期吸入才有效果。气体成分及氢气比例不固定，有氢气、氮气混合气体，家用氢气吸入器产生的是氢氧混合气体，氢氧比例为 2：1，也有氢气发生器能分离氧气，产生高纯度氢气。在体内实验中，氢气吸入比例不能超过爆炸极限 4%，否则实际操作时会有危险。富氢水饱和浓度低（0.8mmol/L），不方便储存，必须大剂量给药，难以应用于吞咽困难、不适宜大量输注的患者。另外，富氢水的质量标准和储存困难也限制了它的应用。目前，制备氢水的方法很多，用于氢医学研究的富氢水缺乏统一的质量标准。生产富氢水的方法主要有鼓泡法和高压溶解法。所用的溶剂包括纯净水、生理盐水，甚至器官保存液（Celsior）[ 86 ] . There are also methods of producing hydrogen-rich electrolytic water by electrolysis of water, or using hydrogen rods to produce hydrogen-rich water by chemically reacting magnesium with water. In addition to differences in hydrogen concentration, hydrogen-rich water obtained by these methods also differs in the types of electrolytes, ionic strength, redox potential, pH, etc., which may affect the therapeutic effect. For specific diseases, inhaling hydrogen and drinking hydrogen-rich water are more suitable for the treatment of respiratory diseases and gastrointestinal diseases, but they are difficult to meet the treatment needs of remote tissue diseases such as brain diseases and solid tumors. Yamamoto et al.
  还有通过电解水来制备富氢电解水的方法，或者利用氢气棒通过镁与水发生化学反应来制备富氢水的方法。这些方法得到的富氢水除了氢气浓度的差异外，电解质的种类、离子强度、氧化还原电位、pH 值等也存在差异，这些差异都可能影响治疗效果。针对具体疾病，吸氢气、喝氢水比较适合治疗呼吸系统疾病、胃肠道疾病，但难以满足脑部疾病、实体肿瘤等远端组织疾病的治疗需求。Yamamoto 等。[ 87 ] studied the biodistribution of hydrogen in SD rats under 3% hydrogen inhalation and found that the highest hydrogen concentration was in the liver, with an equilibrium concentration of 29 mmol/L. However, so far, there is a lack of research data on the hydrogen concentration in tumor tissues. Hydrogen therapy is dependent on hydrogen concentration, but the amount of hydrogen reaching solid tumors is very limited. Hydrogen medical research should focus on the exploration of the hydrogen dose-effect relationship and try to conduct it in a higher concentration range to improve the efficacy.
  研究了 3%氢气吸入条件下 SD 大鼠体内氢气的生物分布，发现氢气浓度最高部位为肝脏，平衡浓度为 29mmol/L。但目前为止，尚缺乏关于肿瘤组织中氢气浓度的研究资料。氢气治疗依赖于氢气浓度，但到达实体肿瘤的氢气量十分有限。氢气医学研究应注重氢气剂量-效应关系的探索，尽量在更高的浓度范围内进行，以提高疗效。

• The nano drug delivery system can effectively solve the shortcomings of gas donor molecules such as poor stability and high toxicity, and realize the controlled release of gas in the body, meeting the tissue specificity and concentration dependence requirements of gas therapy. In the treatment of solid tumors, in order to achieve efficient hydrogen uptake and maximize the anti-cancer effect of hydrogen, hydrogen loading through nanomaterials or in situ hydrogen production at the tumor site has become a new means. For example, the efficient delivery of nano drugs prepared by hydrogen storage materials enhances the anti-tumor effect of hydrogen [ 78-79 ] ; the hydrogen precursor molecule ammonia borane is loaded through nanomaterials and decomposed to release hydrogen in the acidic microenvironment of the tumor [ 80 , 88 ] ; iron nanoparticles can aggregate and react at the tumor site to release hydrogen, inhibiting tumor growth [ 89 ] ; polymer semiconductor photocatalysis in situ hydrogen production is used for hydrogen therapy [ 90 ] . Although electrochemical tumor electrotherapy has been studied for a long time, the physiological effects of hydrogen generated at the cathode during treatment have not been evaluated. Recently, some studies have conducted relevant explorations and proved that electrochemical hydrogen production is effective for tumor treatment in animal models [ 91 ] . These pioneering methods provide new options for hydrogen delivery in vivo, especially for the treatment of malignant tumors. In view of the biosafety advantages of hydrogen, in order to promote the greater therapeutic effect of hydrogen, the safety of the selected carrier material is also worthy of special consideration, and new safety issues should not be introduced. Furthermore, whether the changes in the tumor microenvironment caused by chemical reactions have an impact on the physiological effects of hydrogen requires more precise research.
  纳米药物递送系统可有效解决气体供体分子稳定性差、毒性大等缺点，实现气体在体内的可控释放，满足气体治疗的组织特异性和浓度依赖性的要求。在实体肿瘤治疗中，为实现氢气的高效吸收，最大限度发挥氢气的抗癌作用，通过纳米材料载氢或在肿瘤部位原位制氢成为一种新的手段。 例如利用储氢材料制备纳米药物的高效递送，增强氢气的抗肿瘤作用 ] ； 将氢前体分子氨硼烷通过纳米材料负载，在肿瘤酸性微环境中分解释放氢气 [ 80,88 ] ； 铁纳米颗粒可以在肿瘤部位聚集并发生反应释放氢气，抑制肿瘤生长 [ 89 ] ；聚合物半导体光催化原位产氢用于氢气治疗 [ 90 ] 。虽然电化学肿瘤电疗研究已久，但治疗过程中阴极产生氢气的生理效应尚未得到评估。 最近，一些研究进行了相关探索，并证明了电化学制氢在动物模型上对肿瘤治疗有效 [ 91 ] 。这些开创性的方法为氢气在体内输送，特别是对恶性肿瘤的治疗提供了新的选择。鉴于氢气的生物安全优势，为了促进氢气发挥更大的治疗作用，所选载体材料的安全性也值得特别考虑，不应引入新的安全问题。此外，化学反应引起的肿瘤微环境变化是否对氢气的生理效应产生影响还需要更精准的研究。

• In addition to conventional methods of inhaling hydrogen and drinking hydrogen-rich water, hydrogen-releasing nanomaterials, such as coral calcium, or other hydrogen-producing materials can also be taken orally
  除了传统的吸入氢气、饮用富氢水的方法外，还可以通过口服释放氢气的纳米材料，如珊瑚钙或其他产氢材料[ 64 , 92 ] . In addition, some studies have treated a variety of diseases by intraperitoneal injection of hydrogen-containing saline
  此外，一些研究通过腹腔注射含氢盐水来治疗多种疾病[ 93-95 ] . Hydrogen-containing saline eye drops have been used for retinal ischemia-reperfusion injury and corneal alkali burns, with good results
  含氢盐水滴眼液用于治疗视网膜缺血再灌注损伤、角膜碱烧伤，效果良好[ 96-97 ] . In skin disease research, hydrogen intake through hydrogen water baths has also become an effective method
  在皮肤病研究中，通过氢水浴摄入氢气也成为一种有效的方法[ 98 ] . These methods are worth learning from in the hydrogen treatment of tumors in specific locations. Magnesium reacts with water to produce hydrogen, which is often used in anti-inflammatory treatment research. Metal magnesium stents used in orthopedics also produce hydrogen during degradation. Based on this principle, Zan et al.
  氢气治疗特定部位的肿瘤。镁与水反应产生氢气，常用于抗炎治疗研究。骨科使用的金属镁支架在降解过程中也会产生氢气。基于此原理，Zan 等人研究了氢能治疗特定部位的肿瘤。[ 99 ] used anti-tumor stent materials containing magnesium wire to achieve the combined effect of magnesium ions and hydrogen, inhibiting tumor growth without toxicity to normal tissues, making it an effective hydrogen delivery method for hydrogen anti-tumor treatment. They also regulated heterogeneous metals to accelerate the release of hydrogen from high-purity magnesium, enhancing the inhibitory effect of hydrogen on the proliferation of thyroid tumor Cal-62 cells
  利用含镁丝的抗肿瘤支架材料，实现镁离子与氢气的联合作用，抑制肿瘤生长，且对正常组织无毒，是氢气抗肿瘤治疗的一种有效氢气输送方式。他们还调控异质金属加速高纯镁释放氢气，增强氢气对甲状腺肿瘤 Cal-62 细胞增殖的抑制作用[ 100 ] . Hydrogen itself is an endogenous gas. Intestinal anaerobic microorganisms produce up to 12L of hydrogen per day, most of which is converted into hydrogen sulfide, methane and acetic acid by bacteria in the intestine. The final exhaled hydrogen concentration can reach 5-10ppm
  氢气本身是一种内源性气体，肠道厌氧微生物每天产生氢气高达 12L，大部分被肠道细菌转化为硫化氢、甲烷和醋酸，最终呼出的氢气浓度可达 5-10ppm[ 101 ] . The hydrogen breath test (HBT) is used to measure the concentration of hydrogen in human exhaled gas, which has become a method for detecting gastrointestinal function
  氢气呼气试验（HBT）是用于测量人体呼出气体中氢气浓度的一种方法，已成为检测胃肠功能的一种方法[ 102 ] . The hydrogen ingested by breathing, drinking or injection is far lower than the hydrogen production of the gastrointestinal tract, but hydrogen supplementation has a significant protective effect. There is a contradiction between the human body's ability to generate hydrogen and the need to ingest exogenous hydrogen for disease treatment, which requires further study. In addition, inducing the gastrointestinal tract (such as ingesting acarbose, fructose, turmeric, milk, etc.) to promote intestinal microbial hydrogen production can help treat some diseases. For example, Kajiya et al.
  通过呼吸、饮用或注射摄入的氢气远低于胃肠道的产氢量，但补充氢气有明显的保护作用。人体产氢能力与疾病治疗需要摄取外源性氢气之间存在矛盾，有待进一步研究。此外，诱导胃肠道(如摄取阿卡波糖、果糖、姜黄、牛奶等)促进肠道微生物产氢，有助于治疗一些疾病。例如，Kajiya 等。[ 103 ] supplemented animals with hydrogen-producing bacteria to induce the gastrointestinal tract to produce more hydrogen, which had a therapeutic effect on concanavalin A-induced hepatitis. However, this effect completely disappeared when antibiotics were used to kill bacteria. However, since the hydrogen produced is limited to the digestive tract, the types of diseases that can be applied are limited and are affected by individual physiological conditions
  给动物补充产氢菌，诱导胃肠道产生更多氢气，对刀豆球蛋白 A 诱发的肝炎有治疗作用。但当使用抗生素杀灭细菌时，这种作用就完全消失了。但由于产生的氢气仅限于消化道，因此可应用的疾病类型有限，且受个体生理状况影响[ 86 ] . In the field of hydrogen treatment of tumors, these methods have important reference value.
  在氢气治疗肿瘤领域，这些方法具有重要的参考价值。

• 6 Summary 6 总结

• At present, with the clinical application of hydrogen in respiratory diseases such as the new coronavirus pneumonia, the safety and medical effects of hydrogen have been effectively verified. Clinical research on hydrogen in the treatment of other diseases is also being widely carried out. my country's hydrogen medicine is in a leading position internationally. With the development and standardization of related hydrogen inhalation equipment and the in-depth research on hydrogen in the treatment of different diseases, hydrogen anti-tumor research needs to continue to increase investment. First of all, the top priority of hydrogen anti-tumor research is to discover the exact mechanism of hydrogen anti-tumor. More basic and clinical experiments are needed to further confirm the role of hydrogen and clarify its mechanism of action at the molecular level. The antioxidant mechanism of hydrogen is only one aspect of the action of hydrogen, and there may be other modes of action. At present, the research on direct and indirect signaling pathways mediated by hydrogen molecules is still in the exploratory stage. Whether hydrogen is broad-spectrum anti-tumor or effective for specific tumors needs to be further determined. Secondly, the heterogeneity of tumor tissue puts forward new requirements for hydrogen administration methods, which is also a key factor limiting the efficacy of hydrogen. Therefore, it is also an important research direction to develop a suitable hydrogen delivery system based on the pathological characteristics of tumors, and at the same time, it is necessary to find out the dose-effect relationship of hydrogen as soon as possible. In conclusion, hydrogen has great potential in the field of cancer prevention and treatment and deserves further study.
  目前，随着氢气在新型冠状病毒肺炎等呼吸系统疾病的临床应用，氢气的安全性和医疗效果得到有效验证，氢气在其他疾病治疗中的临床研究也正在广泛开展。我国氢气医学在国际上处于领先地位，随着相关氢气吸入设备的研制和标准化，以及氢气在不同疾病治疗中的深入研究，氢气抗肿瘤研究还需继续加大投入。首先，氢气抗肿瘤研究的当务之急是发现氢气抗肿瘤的确切机制，需要更多的基础和临床实验来进一步证实氢气的作用，在分子水平上明确其作用机制。氢气的抗氧化机制只是氢气作用的一个方面，还可能存在其他的作用方式。目前对氢气分子介导的直接和间接信号通路的研究还处于探索阶段，氢气是否具有广谱抗肿瘤作用或对特定肿瘤有效还有待进一步确定。其次，肿瘤组织的异质性对氢气给药方式提出了新的要求，这也是限制氢气疗效的关键因素。因此，针对肿瘤的病理特点研制合适的氢气输送系统也是一个重要的研究方向，同时需要尽快弄清氢气的量效关系。总之，氢气在癌症防治领域具有巨大的潜力，值得进一步研究。

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