富氢水对结直肠癌荷瘤小鼠肿瘤的生长抑制作用及其机制

Inhibitory effects of hydrogen-rich water against tumor growth in tumor-bearing mice with colorectal cancer and the molecular mechanisms

目的初步探讨富氢水(hydrogen-rich water,[H]H2O)对结直肠癌荷瘤小鼠肿瘤的抑制作用及其机制。方法小鼠右腋皮下接种人结直肠癌SW480细胞(4×106/只),建立移植瘤小鼠模型,造模的同时给予小鼠自由饮用富氢水(0.8mg/kg),每日早、中、晚3次更换新鲜富氢水。对照组给予普通水。荷瘤28d后处死小鼠,取肿瘤组织,通过检测肿瘤体积和质量判断肿瘤的生长情况。用免疫组化法检测肿瘤组织中Ki-67蛋白的表达,TUNEL法检测细胞凋亡情况,比色法检测谷胱甘肽(GSH)及氧化型谷胱甘肽(GSSG)水平,基因芯片和qRT-PCR方法筛选并验证差异表达基因。结果与对照组相比,富氢水干预后可抑制小鼠肿瘤的形成,减小肿瘤体积和质量(P<0.01,P<0.05),降低Ki-67蛋白的表达(P<0.05),促进细胞凋亡(P<0.05),增高GSSG/GSH比值(P<0.05);富氢水干预可调节γ-干扰素(IFN-γ)相关基因的表达。结论在结直肠癌中,富氢水可通过抑制细胞增殖、促进细胞凋亡、调节细胞内氧化还原环境及IFN-γ相关基因的表达而发挥抗肿瘤活性作用。

Objective To investigate the inhibitory effects of hydrogen-rich water（[H]H2O）against tumor growth in mice with colorectal cancer and the related molecular mechanisms.Methods Mice were vaccinated with SW480 cells suspension（4×106 cells/mice）to establish mouse model of colon cancer.The mice in [H]H2O intervention group were freely allowed to drink [H]H2O（0.8mg/kg）,which was refreshed three times daily,and the mice in control group were given light water.The tumor weight and volume were measured 28 dafter bearing tumor to assess tumor growth,the expression of Ki-67 protein was examined by immunohistochemical method,the apoptosis of tumor cells was observed by TUNEL,and the levels of oxidized glutathione（GSSG）and glutathione（GSH）were analyzed using colorimetric method.Micro array was used to screen the differentially expressed genes between control group and [H]H2O intervention group,and the levels of those genes were verified with qRT-PCR.Results Compared with the control group,[H]H2O intervention significantly inhibited the tumor growth,reduced tumor weight and volume（P〈0.01,P〈0.05）;moreover,it also reduced the expression of Ki-67protein（P〈0.05）and promoted tumor cell apoptosis（P〈0.05）.In addition,the ratio of GSSG/GSH was significantly increased in[H]H2O intervention group compared with that in the control group（P〈0.05）.[H]H2O intervention could also regulate the expression of interferonγ-related genes.Conclusion [H]H2O may exert its antitumor activity via inhibiting proliferation,inducing apoptosis of colon cancer cells,and regulating the intracellular redox state and the expression of interferonγ-related genes.