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. 2014 May;18(5):938-46.
doi: 10.1111/jcmm.12245. Epub 2014 Mar 12.

Protective Effect of Hydrogen-Rich Saline Against Radiation-Induced Immune Dysfunction

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Free PMC article

Protective Effect of Hydrogen-Rich Saline Against Radiation-Induced Immune Dysfunction

Sanhu Zhao et al. J Cell Mol Med. .
Free PMC article

Abstract

Recent studies showed that hydrogen can be used as an effective radioprotective agent through scavenging free radicals. This study was undertaken to evaluate the radioprotective effects of hydrogen on immune system in mice. H(2) was dissolved in physiological saline using an apparatus produced by our department. Spleen index and histological analysis were used to evaluate the splenic structural damage. Spleen superoxide dismutase, GSH, MDA were measured to appraise the antioxidant capacity and a DCF assay for the measurement of radical oxygen species. Cell apoptosis was evaluated by an Annexin V-FITC and propidium iodide staining method as well as the apoptotic proteins such as Bcl-2, Bax, caspase-3 and c-caspase-3. CD4+ and CD8+ T cells subtypes were detected by flow cytometry with FITC-labelled antimouse CD4 and PE antimouse CD8 staining. Real-time PCR was utilized to determine the CD4+ T cell subtypes and related cytokines. Our study demonstrated that pre-treatment with H(2) could increase the spleen index and attenuate the radiation damage on splenic structure. Radical oxygen species level was also reduced by H(2) treatment. H(2) also inhibited radiation-induced apoptosis in splenocytes and down-regulated pro-apoptotic proteins in living mice. Radiation-induced imbalance of T cells was attenuated by H(2). Finally, we found that H(2) could regulate the polarization of CD4+ T cells and the level of related cytokines. This study suggests H(2) as an effective radioprotective agent on immune system by scavenging reactive oxygen species.

Keywords: apoptosis; hydrogen; immune regulation; ionizing radiation; radioprotection.

Figures

Fig. 1
Fig. 1
Spleen index 14 days after irradiation. Spleen index = spleen weight/bodyweight × 100. The data were expressed as means ± SEM (n = 8), *P < 0.05.
Fig. 2
Fig. 2
Morphology of spleen after in vivo γ-radiation. (A) Spleens of mice with control morphology are comprised of both red and white pulps. (B) Spleens of mice 2 days after in vivo γ-radiation. (C) Spleens in mice pre-treated with H2 at 2 days after irradiation. (D) Spleens of mice 14 days after in vivo γ-radiation. (E) Spleens in mice pre-treated with H2 at 14 days after irradiation.
Fig. 3
Fig. 3
Effects of hydrogen to antioxidant indexes. Changes in (A) levels of spleen superoxide dismutase and (B) activities of GSH, and (C) concentration of MDA in normal, radiation and H2-pre-treated mice. (D) DCF-DA fluorescence images were obtained at 2, 4, 6 hours after 5 Gy radiation. (F) DCFH-DA fluorescence intensity was semiquantified from the cryosection of each independent experiment. The data were expressed as means ± SEM (n = 8), *P < 0.05.
Fig. 4
Fig. 4
Hydrogen attenuated irradiation-induced splenocytes apoptosis. (A and B) Irradiation-induced splenocytes apoptosis rate was quantified by Flow Cytometry at 24, 36, 48 hrs after exposure. (C) Bcl-2, Bax, caspase-3, c-caspase-3 protein expression was detected by western blot at 14 days after exposure. (D) Normalization of Bcl-2, Bax, caspase-3, c-caspase-3 expression to Tubulin. The data were expressed as means ± SEM (n = 8), *P < 0.05.
Fig. 5
Fig. 5
Effects of hydrogen on radiation-induced disorder in T cell. (A) The total number of splenocytes in normal, radiation, H2-pre-treated mice 14 days after exposure. (B and C) The absolute number and percentage of CD4 + and CD8 + T cells in normal, radiation, H2-pre-treated mice 14 days after exposure. (DG) T-bet, GATA-3, Foxp-3, RORγt mRNA expression was detected by real-time PCR and normalized to GAPDH. The data were expressed as means ± SEM (n = 8), *P < 0.05.
Fig. 6
Fig. 6
Effects of hydrogen on mRNA expression of Th-type cytokines and pro-inflammatory cytokines in vivo. (A and B) Effects of hydrogen on mRNA expression of Th1-type cytokines in vivo. (CE) Effects of hydrogen on mRNA expression of Th2-type cytokines in vivo. (F and G) Effects of hydrogen on mRNA expression of Th17-type cytokines in vivo. (H) Effects of hydrogen on mRNA expression of Treg-type cytokines in vivo. (IK) Effects of hydrogen on mRNA expression of pro-inflammatory cytokines in vivo. All those data were normalized to GAPDH. The data were expressed as means ± SEM (n = 8), *P < 0.05.

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References

    1. Oradovskaia IV, Pashchenkova IuG, Feoktistov VV, et al. The epidemiological analysis of monitoring of the immune status in liquidators of consequences of the Chernobyl accident for early identification of risk groups and diagnostics of oncological diseases. Report 2. Dependence of frequency and changes in the immune status on risk factors of radiation accident. Radiats Biol Radioecol. 2011;51:117–33. - PubMed
    1. Borja-Aburto VH, Bustamante-Montes P, Garcia-Sancho MC, et al. Ionizing radiation at low doses and cancer: epidemiological controversy. Rev Invest Clin. 1990;42:312–6. - PubMed
    1. Makidono R, Ouchida T, Makidono A, et al. Severe damage of CD4-2H4+ T subpopulation cells (naive T cells and suppressor/inducer) by radiation therapy, their recovery being promoted by a plant alkaloid. Nihon Igaku Hoshasen Gakkai Zasshi. 1992;52:223–8. - PubMed
    1. Dainiak N, Waselenko JK, Armitage JO, et al. The hematologist and radiation casualties. Hematology Am Soc Hematol Educ Program. 2003;1:473–96. - PubMed
    1. Hayashi T, Morishita Y, Kubo Y, et al. Long-term effects of radiation dose on inflammatory markers in atomic bomb survivors. Am J Med. 2005;118:83–6. - PubMed

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