Journal of Zhejiang University SCIENCE B

, Volume 14, Issue 5, pp 382–391 | Cite as

Protective effect of hydrogen-rich saline on ischemia/reperfusion injury in rat skin flap

  • Ling Zhao
  • You-bin Wang
  • Shi-rui Qin
  • Xue-mei Ma
  • Xue-jun Sun
  • Ming-lian Wang
  • Ru-gang Zhong



Skin damage induced by ischemia/reperfusion (I/R) is a multifactorial process that often occurs in plastic surgery. The mechanisms of I/R injury include hypoxia, inflammation, and oxidative damage. Hydrogen gas has been reported to alleviate cerebral I/R injury by acting as a free radical scavenger. Here, we assessed the protective effect of hydrogen-rich saline (HRS) on skin flap I/R injury.


Abdominal skin flaps of rats were elevated and ischemia was induced for 3 h; subsequently, HRS or physiological saline was administered intraperitoneally 10 min before reperfusion. On postoperative Day 5, flap survival, blood perfusion, the accumulation of reactive oxygen species (ROS), and levels of cytokines were evaluated. Histological examinations were performed to assess inflammatory cell infiltration.


Skin flap survival and blood flow perfusion were improved by HRS relative to the controls. The production of malondialdehyde (MDA), an indicator of lipid peroxidation, was markedly reduced. A multiplex cytokine assay revealed that HRS reduced the elevation in the levels of inflammatory cytokines, chemokines and growth factors, with the exception of RANTES (regulated on activation, normal T-cell expressed and secreted) growth factor. HRS treatment also reduced inflammatory cell infiltration induced by I/R injury.


Our findings suggest that HRS mitigates I/R injury by decreasing inflammation and, therefore, has the potential for application as a therapy for improving skin flap survival.

Key words

Hydrogen-rich saline Ischemia/reperfusion Skin flaps Oxidative damage Inflammation 

CLC number

Q74 R459.9 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Arato, E., Kurthy, M., Sinay, L., Kasza, G., Menyhei, G., Hardi, P., Masoud, S., Ripp, K., Szilagyi, K., Takacs, I., et al., 2010. Effect of vitamin E on reperfusion injuries during reconstructive vascular operations on lower limbs. Clin. Hemorheol. Microcirc., 44(2):125–136. [doi:10.3233/CH-2010-1260]PubMedGoogle Scholar
  2. Aydogan, H., Gurlek, A., Parlakpinar, H., Askar, I., Bay-Karabulut, A., Aydogan, N., Fariz, A., Acet, A., 2007. Beneficial effects of caffeic acid phenethyl ester (CAPE) on the ischaemia-reperfusion injury in rat skin flaps. J. Plast. Reconstr. Aesthet. Surg., 60(5):563–568. [doi:10.1016/j.bjps.2006.01.030]PubMedCrossRefGoogle Scholar
  3. Buchholz, B.M., Kaczorowski, D.J., Sugimoto, R., Yang, R., Wang, Y., Billiar, T.R., McCurry, K.R., Bauera, A.J., Nakao, A., 2008. Hydrogen inhalation ameliorates oxidative stress in transplantation induced intestinal graft injury. Am. J. Transplant., 8(10):2015–2024. [doi:10.1111/j.1600-6143.2008.02359.x]PubMedCrossRefGoogle Scholar
  4. Chen, H., Sun, Y.P., Hu, P.F., Liu, W.W., Xiang, H.G., Li, Y., Yan, R.L., Su, N., Ruan, C.P., Sun, X.J., et al., 2011. The effects of hydrogen-rich saline on the contractile and structural changes of intestine induced by ischemia-reperfusion in rats. J. Surg. Res., 167(2):316–322. [doi:10.1016/j.jss.2009.07.045]PubMedCrossRefGoogle Scholar
  5. Dacho, A., Lyutenski, S., Aust, G., Dietz, A., 2009. Ischemic preconditioning in a rat adipocutaneous flap model. HNO, 57(8):829–834 (in German). [doi:10.1007/s00106-009-1901-8]PubMedCrossRefGoogle Scholar
  6. Fukuda, K., Asoh, S., Ishikawa, M., Yamamoto, Y., Ohsawa, I., Ohta, S., 2007. Inhalation of hydrogen gas suppresses hepatic injury caused by ischemia/reperfusion through reducing oxidative stress. Biochem. Biophys. Res. Commun., 361(3):670–674. [doi:10.1016/j.bbrc.2007.07.088]PubMedCrossRefGoogle Scholar
  7. Gurlek, A., Celik, M., Parlakpinar, H., Aydogan, H., Bay-Karabulut, A., 2006. The protective effect of melatonin on ischemia-reperfusion injury in the groin (inferior epigastric) flap model in rats. J. Pineal Res., 40(4):312–317. [doi:10.1111/j.1600-079X.2006.00319.x]PubMedCrossRefGoogle Scholar
  8. Harder, Y., Amon, M., Laschke, M.W., Schramm, R., Rucker, M., Wettstein, R., Bastiaanse, J., Frick, A., Machens, H.G., Küntscher, M., et al., 2008. An old dream revitalised: preconditioning strategies to protect surgical flaps from critical ischemia and ischaemia-reperfusion injury. J. Plast. Reconstr. Aesthet. Surg., 61(5):503–511. [doi:10.1016/j.bjps.2007.11.032]PubMedCrossRefGoogle Scholar
  9. Hayashida, K., Sano, M., Ohsawa, I., Shinmura, K., Tamaki, K., Kimura, K., Endo, J., Katayama, T., Kawamura, A., Kohsaka, S., et al., 2008. Inhalation of hydrogen gas reduces infarct size in the rat model of myocardial ischemia-reperfusion injury. Biochem. Biophys. Res. Commun., 373(1):30–35. [doi:10.1016/j.bbrc.2008.05.165]PubMedCrossRefGoogle Scholar
  10. Henderson, P.W., Singh, S.P., Weinstein, A.L., Nagineni, V., Rafii, D.C., Kadouch, D., Krijgh, D.D., Spector, J.A., 2010. Therapeutic metabolic inhibition: hydrogen sulfide significantly mitigates skeletal muscle ischemia reperfusion injury in vitro and in vivo. Plast. Reconstr. Surg., 126(6):1890–1898. [doi:10.1097/PRS.0b013e3181f446bc]PubMedCrossRefGoogle Scholar
  11. Imai, S., Kozai, H., Matsuda, M., Hasegawa, G., Obayashi, H., Togawa, C., Yamamura, T., Watanabe, K., Miyatani, S., Yoshikawa, T., et al., 2008. Intervention with delivery of diabetic meals improves glycemic control in patients with type 2 diabetes mellitus. J. Clin. Biochem. Nutr., 42(1): 59–63. [doi:10.3164/jcbn.2008010]PubMedCrossRefGoogle Scholar
  12. Ito, M., Ibi, T., Sahashi, K., Ichihara, M., Ito, M., Ohno, K., 2011. Open-label trial and randomized, double-blind, placebo-controlled, crossover trial of hydrogen-enriched water for mitochondrial and inflammatory myopathies. Med. Gas Res., 1(1):24. [doi:10.1186/2045-9912-1-24]PubMedCrossRefGoogle Scholar
  13. Kajiya, M., Sato, K., Silva, M.J., Ouhara, K., Do, P.M., Shanmugam, K.T., Kawai, T., 2009. Hydrogen from intestinal bacteria is protective for concanavalin A-induced hepatitis. Biochem. Biophys. Res. Commun., 386(2): 316–321. [doi:10.1016/j.bbrc.2009.06.024]PubMedCrossRefGoogle Scholar
  14. Kang, K.M., Kang, Y.N., Choi, I.B., Gu, Y., Kawamura, T., Toyoda, Y., Nakao, A., 2011. Effects of drinking hydrogen-rich water on the quality of life of patients treated with radiotherapy for liver tumors. Med. Gas Res., 1(1):11. [doi:10.1186/2045-9912-1-11]PubMedCrossRefGoogle Scholar
  15. Karaaslan, O., Ulusoy, M.G., Kankaya, Y., Tiftikcioglu, Y.O., Kocer, U., Kankaya, D., Karaaslan, G.M., Tuncer, S., Berktas, M., 2010. Protective effect of grape seed extract against ischaemia/reperfusion injury in a rat epigastricflap model. J. Plast. Reconstr. Aesthet. Surg., 63(4): 705–710. [doi:10.1016/j.bjps.2009.01.018]PubMedCrossRefGoogle Scholar
  16. Küntscher, M.V., Schirmbeck, E.U., Menke, H., Klar, E., Gebhard, M.M., Germann, G., 2002a. Ischemic preconditioning by brief extremity ischemia before flap ischemia in a rat model. Plast. Reconstr. Surg., 109(7): 2398–2404. [doi:10.1097/00006534-200206000-00034]PubMedCrossRefGoogle Scholar
  17. Küntscher, M.V., Juran, S., Menke, H., Gebhard, M.M., Erdmann, D., Germann, G., 2002b. The role of pre-ischaemic application of the nitric oxide donor spermine/nitric oxide complex in enhancing flap survival in a rat model. Br. J. Plast. Surg., 55(5):430–433. [doi:10.1054/bjps.2002.3871]PubMedCrossRefGoogle Scholar
  18. Manson, P.N., Anthenelli, R.M., Im, M.J., Bulkley, G.B., Hoopes, J.E., 1983. The role of oxygen-free radicals in ischemic tissue injury in island skin flaps. Ann. Surg., 198(1):87–90. [doi:10.1097/00000658-198307000-00017]PubMedCrossRefGoogle Scholar
  19. Moon, J.G., Lim, H.C., Gye, M.R., Oh, J.S., Park, J.W., 2008. Postconditioning attenuates ischemia-reperfusion injury in rat skin flap. Microsurgery, 28(7):531–537. [doi:10.1002/micr.20530]PubMedCrossRefGoogle Scholar
  20. Nagata, K., Nakashima-Kamimura, N., Mikami, T., Ohsawa, I., Ohta, S., 2009. Consumption of molecular hydrogen prevents the stress-induced impairments in hippocampus-dependent learning tasks during chronic physical restraint in mice. Neuropsychopharmacology, 34(2):501–508. [doi:10.1038/npp.2008.95]PubMedCrossRefGoogle Scholar
  21. Nakao, A., Sugimoto, R., Billiar, T.R., McCurry, K.R., 2009. Therapeutic antioxidant medical gas. J. Clin. Biochem. Nutr., 44(1):1–13. [doi:10.3164/jcbn.08-193R]PubMedCrossRefGoogle Scholar
  22. Nakayama, M., Nakano, H., Hamada, H., Itami, N., Nakazawa, R., Ito, S., 2010. A novel bioactive haemodialysis system using dissolved dihydrogen (H2) produced by water electrolysis: a clinical trial. Nephrol. Dial. Transplant., 25(9):3026–3033. [doi:10.1093/ndt/gfq196]PubMedCrossRefGoogle Scholar
  23. 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(6):688–694. [doi:10.1038/nm1577]PubMedCrossRefGoogle Scholar
  24. Ono, H., Nishijima, Y., Adachi, N., Sakamoto, M., Kudo, Y., Nakazawa, J., Kaneko, K., Nakao, A., 2012. Hydrogen (H2) treatment for acute erythymatous skin diseases. A report of 4 patients with safety data and a non-controlled feasibility study with H2 concentration measurement on two volunteers. Med. Gas Res., 2(1):14. [doi:10.1186/2045-9912-2-14]PubMedCrossRefGoogle Scholar
  25. Siemionow, M., Arslan, E., 2004. Ischemia/reperfusion injury: a review in relation to free tissue transfers. Microsurgery, 24(6):468–475. [doi:10.1002/micr.20060]PubMedCrossRefGoogle Scholar
  26. Sun, H., Chen, L., Zhou, W., Hu, L., Li, L., Tu, Q., Chang, Y., Liu, Q., Sun, X., Wu, M., et al., 2011. The protective role of hydrogen-rich saline in experimental liver injury in mice. J. Hepatol., 54(3):471–480. [doi:10.1016/j.jhep. 2010.08.011]PubMedCrossRefGoogle Scholar
  27. van den Heuvel, M.G., Buurman, W.A., Bast, A., van der Hulst, R.R., 2009. Review: ischaemia-reperfusion injury in flap surgery. J. Plast. Reconstr. Aesthet. Surg., 62(6):721–726. [doi:10.1016/j.bjps.2009.01.060]PubMedCrossRefGoogle Scholar
  28. Xie, K., Yu, Y., Zhang, Z., Liu, W., Pei, Y., Xiong, L., Hou, L., Wang, G., 2010. Hydrogen gas improves survival rate and organ damage in zymosan-induced generalized inflammation model. Shock, 34(5):495–501. [doi:10.1097/SHK. 0b013e3181def9aa]PubMedCrossRefGoogle Scholar
  29. Yan, H., Zhang, F., Kochevar, A.J., Akdemir, O., Gao, W., Angel, M., 2010. The effect of postconditioning on the muscle flap survival after ischemia-reperfusion injury in rats. J. Invest. Surg., 23(5):249–256. [doi:10.3109/0894 1931003615529]PubMedCrossRefGoogle Scholar
  30. Zheng, X., Mao, Y., Cai, Y., Li, Y., Liu, W., Sun, P., Zhang, J.H., Sun, X., Yuan, H., 2009. Hydrogen-rich saline protects against intestinal ischemia/reperfusion injury in rats. Free Radic. Res., 43(5):478–484. [doi:10.1080/10715760 902870603]PubMedCrossRefGoogle Scholar

Copyright information

© Zhejiang University and Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  1. 1.College of Life Science and BioengineeringBeijing University of TechnologyBeijingChina
  2. 2.Peking Union Medical College HospitalBeijingChina
  3. 3.Department of Diving Medicine, Faculty of Naval Medicinethe Second Military Medical UniversityShanghaiChina

Personalised recommendations