果蔬发酵液对小鼠酒精性肝损伤的保护作用及氧化应激机制  

THE PROTECTIVE EFFECT OF VEGETABLES AND FRUITS FERMENTATION LIQUID IN MICE WITH ALCOHOLIC LIVER DAMAGE AND ITS ANTIOXIDATIVE MECHANISM

作　　者：谷大为 陈志敏[1] 周明[1] 陆路[2] 谷瑞增[2] 蔡云清[1] 

机构地区：[1]南京医科大学公共卫生学院营养与食品卫生系,南京211166 [2]中国食品发酵工业研究院,北京100015

出　　处：《营养学报》2015年第4期366-371,共6页Acta Nutrimenta Sinica

基　　金：江苏高校优势学科建设工程资助项目(2011)

摘　　要：目的研究果蔬发酵液(vegetables and fruits ferment liquid,VFFL)对小鼠酒精性肝损伤的保护作用及其氧化应激机制。方法 50只雄性ICR小鼠随机分为正常对照组、乙醇模型组、果蔬发酵液组[10、30、60ml/(kg·d)],正常对照组和乙醇模型组蒸馏水灌胃,其余组用不同剂量果蔬发酵液灌胃,连续30d,最后一次灌胃16h后,乙醇模型组和各果蔬发酵液组均1次性50%乙醇12ml/kg·bw灌胃,6h后处死、取血和肝组织。分别采用HE染色观察肝组织,试剂盒测定血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)、碱性磷酸酶(AKP)含量及肝组织超氧化物歧化酶(SOD)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)、蛋白质羰基、活性氧(ROS)含量,TUNEL法测定肝细胞凋亡,蛋白免疫印迹法检测凋亡相关蛋白表达。结果与乙醇模型组比较,果蔬发酵液组的肝组织水肿和坏死明显减轻;血清中的ALT、AST、AKP水平和肝细胞凋亡明显低于乙醇模型组(P<0.05);肝组织中SOD、GSH、GSH-Px水平明显高于乙醇模型组(P<0.05),而ROS、MDA、蛋白质羰基含量明显低于乙醇模型组(P<0.05);果蔬发酵液组的VDAC(电压依赖性阴离子通道蛋白)、Bcl-2蛋白表达增高(P<0.05),细胞色素C、caspase-9、caspase-3、Bax蛋白表达下降(P<0.05)。结论果蔬发酵液具有较好的抗氧化作用,它可通过阻止ROS介导的线粒体信号通路,抑制肝细胞凋亡,从而保护小鼠酒精性肝损伤。ObjectiveTo investigate the protective effect of vegetables and fruits ferment liquid （VFFL） in mice with alcohol damage and its mechanism.Methods A total of 50 male ICR mice were randomly divided into 5 groups, including normal control group, model group, VFFL groups （10,30,60ml/（kg·d））.The VFFL groups were intragastrically administered with corresponding VFFL for 30 days, while the control group and model group was given distilled water.Then, 50% ethanol（12ml/kg·bw） was administered orally to the intervention groups and model groups 16 hour after the last administration.The mice in the normal control group were administered with an equal volume of distilled water.All the mice were sacrificed 6 hours later,and their serum and liver samples were collected.HE （Hematoxylin and Eosin） staining was performed for observing pathological changes of the liver tissues.Hepatic function was evaluated by measuring the serum levels of alanine transaminase （ALT）, aspartate transaminase （AST） and alkaline phosphatase （AKP）. The levels of superoxide dismutase （SOD）, glutathione （GSH） and glutathione peroxidase （GSH-Px） in the liver tissue were measured.We also analyzed the change of malondialdehyde（MDA）, carbonyls and reactive oxygen species（ROS）in the liver. Hepatocyte apoptosis was observed by TdT-mediated dUTP-biotin nick end labeling （TUNEL）.The levels of VDAC, cytochrome C, caspase-9, caspase-3, Bcl-2 and Bax proteins were detected by Western blotting.Results The results of HE staining showed that VFFL significantly improved the liver cell edema and necrosis. Meanwhile, treatments with different doses of VFFL could significantly decrease serum ALT, AST, AKP levels （P〈0.05）. The hepatocyte apoptosis rates in VFFL groups signifi-cantly decreased compared with that in model group（P〈0.01）.Moreover, VFFL significantly reduced the contents of carbonyls, ROS and MDA （P〈0.05）,increased GSH levels, SOD and GSH-Px activities （P〈0.05） in the liver tissue

关 键 词：果蔬发酵液 酒精肝损伤 氧化应激 细胞凋亡 线粒体通路 

分 类 号：R151[医药卫生—营养与食品卫生学]