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双环醇对四环素所致脂肪肝小鼠肝组织增殖指标的作用
作者:姚晓敏 李越 程小艳 胡彧 严琦 
单位:315100  浙江宁波 浙江医药高等专科学校药学院(姚晓敏、胡彧、严琦) 100050  北京 北京市理化分析测试中心(李越、程小艳) 
关键词:双环醇 四环素 脂肪肝 细胞增殖 
分类号:R575.5
出版年,卷(期):页码:2016,41(5):384-388
摘要:
[摘要] 目的 探讨双环醇对四环素所致药物性脂肪肝小鼠肝组织增殖指标的作用。方法 50ICR小鼠随机分为正常组、四环素模型6h组、双环醇给药6h组、四环素模型24h组、双环醇给药24h组,每组10只。给药组给予双环醇300mg/kg,每12h给药1次,共3次,其余各组同时间点给予等量赋型剂;模型组和给药组于末次给药后1h腹腔注射四环素200mg/kg,对照组给予相同pH值等体积生理盐水。各组动物分别于四环素给药后6h24h眼球取血,并取肝组织进行检测。采用实时荧光定量PCR法测定肝组织增殖细胞核抗原(PCNA)、周期素D1(Cyclin D1)c-myc基因表达情况Western blotting法测定肝组织Cyclin D1蛋白表达,免疫组化法测定肝组织PCNAc-myc蛋白的表达。结果 四环素注射后6h小鼠肝细胞PCNA基因表达为对照组的43%(P<0.01),同时蛋白表达显著减少,随着时间延长,至24h时基本恢复,而双环醇给药在脂肪肝早期(6h)即能明显抑制肝细胞PCNA基因和蛋白表达的下降(P<0.01)。四环素注射后24h模型组Cyclin D1表达量为对照组的59%(P<0.01)c-myc表达为对照组的5.7倍,双环醇给药在脂肪肝早期(6h)即可显著促进Cyclin D1c-myc基因表达(P<0.01),至24h时能明显促进Cyclin D1蛋白的表达(P<0.05)结论 双环醇可显著促进四环素所致药物性脂肪肝小鼠肝细胞的增殖,其机制可能与上调增殖基因和蛋白(PCNACyclin D1c-myc)的表达有关。

[Abstract]  Objective  To investigate the effect of bicyclol on liver tissue proliferation in mice with tetracycline-induced fatty liver. Methods  Fifty ICR mice were randomly assigned into 5 groups (10 each): normal group (control), tetracycline 6h group, bicyclol 6h group, tetracycline 24h group and bicyclol 24h group. Mice in the two bicyclol groups received bicyclol (300mg/ kg) by gavage for 3 times in 12h interval, meanwhile, those in the other groups received the equal amount of forming agents. Tetracycline (200mg/kg) was then injected intraperitoneally to the mice in bicyclol groups and tetracycline groups 1h after the last dose of bicyclol, and mice in control group received equal amount and pH of saline. The liver issues and blood samples from eyes were collected 6h and 24h after tetracycline injection for detection of the corresponding indicators. RT-PCR was performed to detect the gene expression of proliferation index (PCNA, cyclin D1 and c-myc), Western blotting was performed to detect the expression of Cyclin D1 in liver tissue, and immunohistochemistry was employed to detect the expression of PCNA and c-myc proteins. Results  The expression level of PCNA gene 6h after tetracycline injection in mice of tetracycline group was 43% of that in control group, meanwhile the expression of protein also reduced obviously. Bicyclol remarkably inhibited the decrease of PCNA gene and protein expressions at early time (6h) of fatty liver. In addition, the expression of cyclin D1 gene in tetracycline 24h group was 59%, and the expression of c-myc was 5.7 folds of that in control group. Bicyclol remarkably up-regulated the gene expressions of cyclin D1 and c-myc 6h after tetracycline injection, and up-regulated the protein expression of cyclin D1 24h after tetracycline injection. Conclusion Bicyclol can enhance hepatocyte proliferation of mice with tetracycline-induced fatty liver by up-regulation of PCNA, cyclin D1 and c-myc expression.

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