Nano-bubble hydrogen water: An effective therapeutic agent against inflammation related disease caused by viral infection in zebrafish model

Chen Li; Yiran Cao; Fukuda Kohei; Haihong Hao; Guiqing Peng; Can Cheng; Jing Ye

doi:10.1016/j.virs.2022.01.023

April 2022

Chen Li, Yiran Cao, Fukuda Kohei, Haihong Hao, Guiqing Peng, Can Cheng and Jing Ye. Nano-bubble hydrogen water: An effective therapeutic agent against inflammation related disease caused by viral infection in zebrafish model[J]. Virologica Sinica, 2022, 37(2): 277-283. doi: 10.1016/j.virs.2022.01.023

Citation: Chen Li, Yiran Cao, Fukuda Kohei, Haihong Hao, Guiqing Peng, Can Cheng, Jing Ye. Nano-bubble hydrogen water: An effective therapeutic agent against inflammation related disease caused by viral infection in zebrafish model .VIROLOGICA SINICA, 2022, 37(2) : 277-283. http://dx.doi.org/10.1016/j.virs.2022.01.023

纳米氢气泡水:一种对病毒感染斑马鱼模型引起的炎症相关疾病的有效治疗制剂

李陈 ^a ,
曹逸然 ^b ,
Fukuda Kohei ^c ,
郝海红 ^d ,
彭贵青 ^e ,
程璨 ^b ,
叶静 ^e,,

a 华中农业大学农业微生物国家重点实验室, 水产学院, 湖北武汉 430070
b 武汉英中高级中学, 湖北武汉 430070
c 深圳纳诺博科技有限公司, 广东深圳 518000
d 华中农业大学国家兽药残留基准实验室, 湖北武汉 430070
e 华中农业大学农业微生物国家重点实验室, 动物医学院, 湖北武汉 430070

通讯作者： 叶静, yej@mail.hzau.edu.cn
收稿日期： 2021-06-09
录用日期： 2021-10-08

摘要

氢气的抗炎作用已广为人知，但氢气微溶于水，导致口服富氢水疗法效果不佳。本研究制备了纳米氢气泡水（nano-HW）（约0.7ppm），并以鲤春病毒血症病毒（SVCV）感染斑马鱼为模型研究其对病毒感染诱导炎症的治疗作用。将3月龄的斑马鱼分为nano-HW处理组和养殖用水处理组（对照组）。结果显示，经纳米氢气泡水处理后，SVCV感染斑马鱼的累积死亡率降低了40%，RT-qPCR结果显示SVCV复制也受到了显著抑制。H&E染色结果显示，经纳米氢气泡水处理后，SVCV感染引起的组织损伤明显减轻。此外，纳米氢气泡水处理后，SVCV感染引起的活性氧（ROS）积累显著减少。体内和体外实验证明，纳米氢气泡水处理组的促炎细胞因子IL-1β、IL-8和TNF-α的水平显著降低。综上所述，我们的数据首次证明纳米氢气泡水可以抑制斑马鱼病毒感染引起的炎症反应，为病毒引起的炎症相关疾病提供了一种新的治疗策略。
- 纳米氢气泡水
- , 鲤春病毒血症病毒
- , 斑马鱼
- , 炎症

Nano-bubble hydrogen water: An effective therapeutic agent against inflammation related disease caused by viral infection in zebrafish model

Chen Li ^a ,
Yiran Cao ^b ,
Fukuda Kohei ^c ,
Haihong Hao ^d ,
Guiqing Peng ^e ,
Can Cheng ^b ,
Jing Ye ^e,,

a State Key Laboratory of Agricultural Microbiology, College of Fisheries, Huazhong Agricultural University, Wuhan, 430070, China
b Wuhan Britain-China School, Wuhan, 430070, China
c Shenzhen Nano Buddy Technology Co., Ltd, Shenzhen, 518000, China
d MOA Laboratory for Risk Assessment of Quality and Safety of Livestock and Poultry Products, Huazhong Agricultural University, Wuhan, 430070, China
e State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, 430070, China

Corresponding author: Jing Ye, yej@mail.hzau.edu.cn
Received Date: 09 June 2021
Accepted Date: 08 October 2021

Abstract

Since the anti-inflammatory effect of hydrogen has been widely known, it was supposed that hydrogen could suppress tissue damage by inhibiting virus-related inflammatory reactions. However, hydrogen is slightly soluble in water, which leads to poor effect of oral hydrogen-rich water therapy. In this study, the nano-bubble hydrogen water (nano-HW) (about 0.7 ppm) was prepared and its therapeutic effect against viral infection was investigated by utilizing spring viraemia of carp virus (SVCV)-infected zebrafish as model. Three-month-old zebrafish were divided into nano-HW treatment-treated group and aquaculture water treated group (control group). The results revealed that the cumulative mortality rate of SVCV-infected zebrafish was reduced by 40% after treatment with nano-bubble hydrogen water, and qRT-PCR results showed that SVCV replication was significantly inhibited. Histopathological examination staining showed that SVCV infection caused tissue damage was greatly alleviated after treatment with nano-bubble hydrogen water. Futhermore, SVCV infection caused reactive oxygen species (ROS) accumulation was significantly reduced upon nano-HW treatment. The level of proinflammatory cytokines IL-1β, IL-8, and TNF-α was remarkably reduced in the nano-HW-treated group in vivo and in vitro. Taken together, our data demonstrated for the first time that nano-HW could inhibit the inflammatory response caused by viral infection in zebrafish, which suggests that nano-HW can be applied to antiviral research,and provides a novel therapeutic strategy for virus-caused inflammation related disease.
- Nano-bubble hydrogen water (nano-HW)
- , Spring viraemia of carp virus (SVCV)
- , Zebrafish
- , Inflammation

References
1. Ahne, W., Bjorklund, H.V., Essbauer, S., Fijan, N., Kurath, G., Winton, J.R., 2002. Spring viremia of carp (SVC). Dis. Aquat. Org. 52, 261–272.
2. Brown, H.K., Schiavone, K., Tazzyman, S., Heymann, D., Chico, T.J., 2017. Zebrafish xenograft models of cancer and metastasis for drug discovery. Expet Opin. Drug Discov. 12, 379–389.
3. Chen, J.B., Kong, X.F., Lv, Y.Y., Qin, S.C., Sun, X.J., Mu, F., Lu, T.Y., Xu, K.C., 2019. Real world survey" of hydrogen-controlled cancer: a follow-up report of 82 advanced cancer patients. Med. Gas Res. 9, 115–121.
4. Chen, J.B., Kong, X.F., Mu, F., Lu, T.Y., Lu, Y.Y., Xu, K.C., 2020. Hydrogen therapy can be used to control tumor progression and alleviate the adverse events of medications in patients with advanced non-small cell lung cancer. Med. Gas Res. 10, 75–80.
5. Dole, M., Wilson, F.R., Fife, W.P., 1975. Hyperbaric hydrogen therapy: a possible treatment for cancer. Science 190, 152–154.
6. Fu, Y., Ito, M., Fujita, Y., Ito, M., Ichihara, M., Masuda, A., Suzuki, Y., Maesawa, S., Kajita, Y., Hirayama, M., Ohsawa, I., Ohta, S., Ohno, K., 2009. Molecular hydrogen is protective against 6-hydroxydopamine-induced nigrostriatal degeneration in a rat model of Parkinson's disease. Neurosci. Lett. 453, 81–85.
7. Guan, W.J., Wei, C.H., Chen, A.L., Sun, X.C., Guo, G.Y., Zou, X., Shi, J.D., Lai, P.Z., Zheng, Z.G., Zhong, N.S., 2020. Hydrogen/oxygen mixed gas inhalation improves disease severity and dyspnea in patients with Coronavirus disease 2019 in a recent multicenter, open-label clinical trial. J. Thorac. Dis. 12, 3448–3452.
8. Hirayama, M., Ito, M., Minato, T., Yoritaka, A., LeBaron, T.W., Ohno, K., 2018. Inhalation of hydrogen gas elevates urinary 8-hydroxy-2'-deoxyguanine in Parkinson's disease. Med. Gas Res. 8, 144–149.
9. Huang, C.S., Kawamura, T., Lee, S., Tochigi, N., Shigemura, N., Buchholz, B.M., Kloke, J.D., Billiar, T.R., Toyoda, Y., Nakao, A., 2010. Hydrogen inhalation ameliorates ventilator-induced lung injury. Crit. Care 14. R234.
10. Iida, A., Nosaka, N., Yumoto, T., Knaup, E., Naito, H., Nishiyama, C., Yamakawa, Y., Tsukahara, K., Terado, M., Sato, K., Ugawa, T., Nakao, A., 2016. The clinical application of hydrogen as a medical treatment. Acta Med. Okayama 70, 331–337.
11. Ishibashi, T., Ichikawa, M., Sato, B., Shibata, S., Hara, Y., Naritomi, Y., Okazaki, K., Nakashima, Y., Iwamoto, Y., Koyanagi, S., Hara, H., Nagao, T., 2015. Improvement of psoriasis-associated arthritis and skin lesions by treatment with molecular hydrogen:a report of three cases. Mol. Med. Rep. 12, 2757–2764.
12. Iuchi, K., Imoto, A., Kamimura, N., Nishimaki, K., Ichimiya, H., Yokota, T., Ohta, S., 2016. Molecular hydrogen regulates gene expression by modifying the free radical chain reaction-dependent generation of oxidized phospholipid mediators. Sci. Rep. 6, 18971.
13. Jackson, K., Dressler, N., Ben-Shushan, R.S., Meerson, A., LeBaron, T.W., Tamir, S., 2018.Effects of alkaline-electrolyzed and hydrogen-rich water, in a high-fat-diet nonalcoholic fatty liver disease mouse model. World J. Gastroenterol. 24, 5095–5108.
14. Kajisa, T., Yamaguchi, T., Hu, A., Suetake, N., Kobayashi, H., 2017. Hydrogen water ameliorates the severity of atopic dermatitis-like lesions and decreases interleukin-1beta, interleukin-33, and mast cell infiltration in NC/Nga mice. Saudi Med. J. 38, 928–933.
15. Kato, S., Saitoh, Y., Miwa, N., 2020. Hydrogen-bubbled platinum-colloid suppresses human esophagus- or tongue-carcinoma cells with intracellular platinum-uptake and the diminished normal-cell mortality. Hum. Cell 33, 1294–1301.
16. Li, J., Dong, Y., Chen, H., Han, H., Yu, Y., Wang, G., Zeng, Y., Xie, K., 2012. Protective effects of hydrogen-rich saline in a rat model of permanent focal cerebral ischemia via reducing oxidative stress and inflammatory cytokines. Brain Res. 1486, 103–111.
17. Liu, W., Shan, L.P., Dong, X.S., Liu, X.W., Ma, T., Liu, Z., 2013. Combined early fluid resuscitation and hydrogen inhalation attenuates lung and intestine injury. World J.Gastroenterol. 19, 492–502.
18. Misk, E., Garver, K., Nagy, E., Isaac, S., Tubbs, L., Huber, P., Al-Hussinee, L., Lumsden, J.S., 2016. Pathogenesis of spring viremia of carp virus in emerald shiner Notropis atherinoides Rafinesque, fathead minnow Pimephales promelas Rafinesque and white sucker Catostomus commersonii (Lacepede). J. Fish. Dis. 39, 729–739.
19. Novoa, B., Figueras, A., 2012. Zebrafish: model for the study of inflammation and the innate immune response to infectious diseases. Adv. Exp. Med. Biol. 946, 253–275.
20. Ohsawa, I., Ishikawa, M., Takahashi, K., Watanabe, M., Nishimaki, K., Yamagata, K., Katsura, K., Katayama, Y., Asoh, S., Ohta, S., 2007. Hydrogen acts as a therapeutic antioxidant by selectively reducing cytotoxic oxygen radicals. Nat. Med. 13, 688–694.
21. Sim, M., Kim, C.S., Shon, W.J., Lee, Y.K., Choi, E.Y., Shin, D.M., 2020. Hydrogen-rich water reduces inflammatory responses and prevents apoptosis of peripheral blood cells in healthy adults: a randomized, double-blind, controlled trial. Sci. Rep. 10, 12130.
22. Wang, Y., Zhang, H., Lu, Y., Wang, F., Liu, L., Liu, J., Liu, X., 2017. Comparative transcriptome analysis of zebrafish (Danio rerio) brain and spleen infected with spring viremia of carp virus (SVCV). Fish Shellfish Immunol. 69, 35–45.
23. Watanabe, M., Kamimura, N., Iuchi, K., Nishimaki, K., Yokota, T., Ogawa, R., Ohta, S., 2017. Protective effect of hydrogen gas inhalation on muscular damage using a mouse hindlimb ischemia-reperfusion injury model. Plast. Reconstr. Surg. 140, 1195–1206.
24. Wu, Y., Yuan, M., Song, J., Chen, X., Yang, H., 2019. Hydrogen gas from inflammation treatment to cancer therapy. ACS Nano 13, 8505–8511.
25. Xiao, L., Miwa, N., 2021. Hydrogen nano-bubble water suppresses ROS generation, adipogenesis, and interleukin-6 secretion in hydrogen-peroxide- or PMA-stimulated adipocytes and three-dimensional subcutaneous adipose equivalents. Cells 10.
26. Xiong, Y., Liu, Y., Cao, L., Wang, D., Guo, M., Jiang, A., Guo, D., Hu, W., Yang, J., Tang, Z., Wu, H., Lin, Y., Zhang, M., Zhang, Q., Shi, M., Liu, Y., Zhou, Y., Lan, K., Chen, Y., 2020. Transcriptomic characteristics of bronchoalveolar lavage fluid and peripheral blood mononuclear cells in COVID-19 patients. Emerg. Microb. Infect. 9, 761–770.
27. Xu, F., Yu, S., Qin, M., Mao, Y., Jin, L., Che, N., Liu, S., Ge, R., 2018. Hydrogen-rich saline ameliorates allergic rhinitis by reversing the imbalance of Th1/Th2 and upregulation of CD4+CD25+Foxp3+Regulatory T cells, interleukin-10, and membranebound transforming growth factor-beta in Guinea pigs. Inflammation 41, 81–92.
28. Yang, M., Dong, Y., He, Q., Zhu, P., Zhuang, Q., Shen, J., Zhang, X., Zhao, M., 2020a. Hydrogen: a novel option in human disease treatment. Oxid. Med. Cell. Longev. 2020, 8384742.
29. Yang, Y., Liu, P.Y., Bao, W., Chen, S.J., Wu, F.S., Zhu, P.Y., 2020b. Hydrogen inhibits endometrial cancer growth via a ROS/NLRP3/caspase-1/GSDMD-mediated pyroptotic pathway. BMC Cancer 20, 28.
30. Yao, Y., Wang, L., Wang, X., 2020. Modeling of solid-tumor microenvironment in zebrafish (Danio rerio) larvae. Adv. Exp. Med. Biol. 1219, 413–428.
31. Zhao, Y.S., An, J.R., Yang, S., Guan, P., Yu, F.Y., Li, W., Li, J.R., Guo, Y., Sun, Z.M., Ji, E.S., 2019. Hydrogen and oxygen mixture to improve cardiac dysfunction and myocardial pathological changes induced by intermittent hypoxia in rats. Oxid. Med. Cell. Longev. 2019, 7415212.
32. Zhuang, K., Zuo, Y.C., Sherchan, P., Wang, J.K., Yan, X.X., Liu, F., 2019. Hydrogen inhalation attenuates oxidative stress related endothelial cells injury after subarachnoid hemorrhage in rats. Front. Neurosci. 13, 1441.
33. Zou, P.F., Nie, P., 2017. Zebrafish as a model for the study of host-virus interactions. Methods Mol. Biol. 1656, 57–78.
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10.1016j.virs.2022.01.023-ESM.docx