Protective effect of hydrogen-rich saline against radiation-induced immune dysfunction. | 放射線による免疫低下に対する水素生理食塩水の予防効果

J Cell Mol Med. 2014 Mar 12. doi: 10.1111/jcmm.12245.

Protective effect of hydrogen-rich saline against radiation-induced immune dysfunction.

Zhejiang Provincial Center for Disease Control and Prevention


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. H2 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 H2 could increase the spleen index and attenuate the radiation damage on splenic structure. Radical oxygen species level was also reduced by H2 treatment. H2 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 H2 . Finally, we found that H2 could regulate the polarization of CD4+ T cells and the level of related cytokines. This study suggests H2 as an effective radioprotective agent on immune system by scavenging reactive oxygen species.

J Cell Mol Med. 2014 Mar 12. doi: 10.1111/jcmm.12245.





研究室自作の装置を用いて、生理食塩水に水素は溶かして水素生理食塩水を作りました。なお、脾臓の損傷を評価するために、Spleen index(脾腫の判定)と組織学的解析を行いました。また、抗酸化能を評価するために脾臓のSOD、GSH、MDAを測定し、活性酸素種量を測定するために、DCFアッセイを行いました。

さらに、アポトーシスを引き起こした細胞の測定にAnnexin V-FITC、PI染色、アポトーシス関与タンパク質(Bcl-2、Bax、カスパーゼ-3とc-カスパーゼ-3)を用いました。

CD4陽性T細胞とCD 8陽性T細胞は、FITC/CD4抗体とPE/CD8抗体を用い、フローメトリーで測定しました。リアルタイムPCR法はCD4陽性T細胞のサプタイプとそれに関与するサイトカインを測定するために用いました。

私たちの研究により、水素を事前に投与することでSpleen indexが改善し、放射線による脾臓の損傷を抑えることがわかりました。また、水素を投与マウスでは活性酸素種の量も減少しました。さらに、水素は放射線によっておこる脾臓細胞のアポトーシスを抑え、生体マウス内でのアポトーシスを引きおこすタンパク質を減少させました。