泮托拉唑对二乙基亚硝胺联合四氯化碳诱导C57BL/6J小鼠肝癌模型肝细胞增殖的影响  

Pantoprazole inhibits cancer cell proliferation in mice with diethylnitrosamine and carbon tetrachloride-induced hepatoma

作　　者：黎元[1] 金海[1] 文国容[1] 庹必光[1] 

机构地区：[1]遵义医学院附属医院消化内科贵州省消化病研究所,贵州省遵义市563000

出　　处：《世界华人消化杂志》2015年第1期22-29,共8页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.81360311~~

摘　　要：目的:探讨泮托拉唑对二乙基亚硝胺(diethylnitrosamine,DEN)联合四氯化碳(carbon tetrachloride,CC14)诱导C57BL/6J小鼠肝癌模型肝癌细胞增殖的影响.方法:将78只C57小鼠采用DEN联合CCl_4共同诱导C57BL/6J小鼠肝癌,第9周将制模小鼠随机分为模型组、泮托拉唑低剂量组(40mg/kg)、高剂量组(80 mg/kg),并开始腹腔注射泮托拉唑,同时取12只正常小鼠为对照.第25周处死小鼠,取正常肝组织、肝癌组织和癌旁组织,肉眼观察拍片,并作HE染色光镜检查.采用生化法检查小鼠血清谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate aminotransferase,AST)、尿素(urea)、肌酐(creatinine,Cr)水平;采用Realtime PCR方法来检测正常肝、肝癌及癌旁组织甲胎蛋白(α-fetoprotein,AFP)、细胞增殖核抗原Ki67,增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)mRNA的表达,采用Western blot法检测Ki67、PCNA蛋白的表达.结果:小鼠死亡率为21%,成瘤率为100%.肝脏组织巨检:模型组与高低剂量组肝体积增大,表面有大小不等的结节,与模型组比较,高、低剂量组结节个数明显减少.HE染色显示:与模型组比较,高、低剂量组异型增生明显减少.血清生化检测显示:模型组、高、低剂量组血清ALT、AST水平明显高于正常组(P<0.05),高剂量组ALT、AST水平明显低于模型组(P<0.05).RT-PCR结果提示,与正常组相比较,模型组和高、低剂量组AFP、Ki67和PCNA mRNA表达明显增高(P<0.05,F=7.94;P<0.05,F=15.62;P<0.01,F=45.58).与模型组比较,高剂量组Ki67和PCNA mRNA表达降低(P<0.05,F=5.89;P<0.05,F=10.34).Western blot结果显示,与正常肝组织比较,模型组和高剂量组的肝癌及癌旁组织Ki67和PCNA蛋白表达明显增高(P<0.05,F=20.41;P<0.05,F=20.31).与模型组比较,高剂量组肝癌及癌旁组织Ki67和PCNA蛋白表达明显减少(P<0.05,F=16.47;P<0.05,F=12.38).结论:泮托拉唑DEN联合CCl_4诱导C57BL/6J小鼠肝癌模型肝细胞增殖有抑制作用.AIM： To investigate the effect of pantoprazole on the proliferation of cancer cells of mice with hepatoma induced by diethylnitrosamine （DEN） plus carbon tetrachloride. METHODS： Seventy-eight C57 mice were treated with DEN and carbon tetrachloride （CCl4） for 9 wk and then randomly divided into three groups： a model group, low- （40 mg/ kg） and high-dose （80 mg/kg） pantoprazole groups. Twelve normal C57 mice were used as controls. The two pantoprazole groups were administrated pantoprazole for 25 wk by intraperitoneal injection, and the mice were then sacrificed for collecting normal liver tissues, hepatoma tissues and hepatoma- adjacent tissues. All these tissues were prepared for pathological observation by HE staining. Blood levels of alanine transaminase （ALT）, aspartate aminotransferase （AST）, urea, and creatinine （Cr） were assayed. The mRNA levels of α-fetoprotein （AFP）, Ki67, and proliferating cell nuclear antigen （PCNA） were quantified by real-time PCR, and the protein levels of Ki67 and PCNA were tested by Western blot. RESULTS： Hepatoma was successfully induced in all mice treated with DEN and carbon tetrachloride, and 21% of the mice were dead. Gross examination of liver tissues showed that the livers in the model group were larger and had more heterogeneous nodules than those in the pantoprazole groups. Pathological analysis showed a lower incidence of dysplasia after administration of pantoprazole. Compared with normal controls, the levels of ALT and AST were increased in the model and pantoprazole groups, and these biological indexes in the pantoprazole groups were significantly lower than those in the model group （P 〈 0.05）. The mRNA levels of AFP, Ki67 and PCNA in the model group and pantoprazole groups were significantly higher than those in normal controls （P 〈 0.05, F = 7.94; P 〈 0.05, F = 15.62; P 〈 0.01, F = 45.58）, and pantoprazole decreased Ki67 and PCNA mRNA levels significantly compared with the model group （P 〈 0.05, F = 5.89

关 键 词：泮托拉唑 二乙基亚硝胺 肝癌 增殖 

分 类 号：R735.7[医药卫生—肿瘤] R-332[医药卫生—临床医学]