甜菜碱对复合致病因素诱导肝硬化大鼠的作用及机制研究  

Protective effects and mechanisms of betaine on liver cirrhosis induced by multiple pathogenic factors in rats

作　　者：田小霞[1] 来丽娜[2] 李旭炯[3] 陈云霞[4] 郭晓姝[3] 

机构地区：[1]长治医学院病理生理学教研室 [2]长治医学院药理学教研室 [3]长治医学院生理学教研室 [4]长治医学院微生物学教研室

出　　处：《中国临床药理学与治疗学》2016年第4期372-377,383,共7页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：山西省高等学校大学生创新创业训练项目(2014308);长治医学院创新团队项目(cx201409)

摘　　要：目的:探讨甜菜碱对复合致病因素诱导肝硬化大鼠的保护作用及可能的机制。方法:72只雄性SD大鼠随机分为空白对照组,甜菜碱干预的对照组,肝硬化模型组和肝硬化模型甜菜碱干预组。各组动物分别于造模第4周末、6周末和8周末处死取材。Elisa法检测大鼠血浆中ALT、内毒素和肝组织匀浆中TNF-α、IL-6水平;肝组织切片行HE染色,NF-κB p65免疫组化染色;实时荧光定量PCR法和Western blot法分别检测肝组织中Toll样受体4(TLR4)mRNA、NF-κB mRNA和TLR4蛋白的表达。结果:与相应的空白对照组相比,肝硬化模型组:血浆中ALT、内毒素水平均随病程进展逐渐显著升高(P<0.05);肝组织中TNF-α、IL-6水平均显著升高(P<0.05),TLR4mRNA和蛋白表达均显著升高(P<0.05),NF-κB mRNA的表达显著升高(P<0.05),NF-κB p65核转位也显著增多(P<0.05),它们的变化趋势为6周水平最高,4周次之,8周最低。与相应的肝硬化模型组相比,肝硬化模型甜菜碱干预组:血浆中ALT、内毒素水平均显著降低(P<0.05);4周和6周组肝组织中TNF-α、IL-6水平均显著降低(P<0.05);肝组织中TLR4 mRNA和蛋白均显著降低(P<0.05),NF-κB mRNA的表达显著降低(P<0.05),NF-κB p65核转位显著降低(P<0.05)。结论:甜菜碱可能通过抑制TLR4/NF-κB信号途径,有效减轻肝脏炎症和纤维化程度,发挥保护肝脏的作用。AIM： To investigate the protective effects and mechanisms of betaine in the development of liver cirrhosis induced by multiple pathogenic factors in rats. METHODS： The seventy-two male SD rats were randomly divided into normal control group（ N）,normal control plus betaine-treated group（ NB）,liver cirrhosis group（ M）,liver cirrhosis plus betaine-treated group（ MB） at the 4th,6th and 8th weeks. Liver cirrhosis in rats was induced by multiple pathogenic factors. Betaine was given in rats of NB and MB groups（ 1g·kg^（-1）·d^（-1）,ig）. The levels of alanine transferase（ ALT）,endotoxin in the plasma and TNF-α,IL-6 in liver tissues were detected by Elisa. Histopathological changes of the liver were observed under microscope with the staining of Hematoxylin and Eosin（ HE）.The expression and localization of nuclear factorkappa B p65（ NF-κB p65） in liver tissues were measured by immunohistochemical staining. The expressions of Toll-like receptors 4（ TLR4） and NF-κB mRNA were detected by real-time fluorescence quantitative polymerase chain reaction（ real-time QPCR）,the expression of TLR4 protein in liver tissues was detected by Western blot. RESULTS：Compared with the corresponding normal control group,liver cirrhosis group： the levels of ALT,endotoxin in the plasma were significantly gradually increased（ P〈 0. 05）; the levels of TNF-α,IL-6 in liver tissues were significantly increased（ P 〈0. 05）,TLR4 mRNA and protein in liver tissues were significantly increased（ P〈 0. 05）,NF-κB mRNA in liver tissues was significantly increased（ P 0. 05）,the percentages of NF-κB p65 translocating to the nucleus significantly increased（ P〈 0. 05）,and they successively increased from the 4th to 6th week and decreased reversely at the 8th week. Compared with the corresponding liver cirrhosis group,liver cirrhosis plus betaine-treated group：the levels of ALT,endotoxin in the plasma were significantly decreased at each time point（ P 〈0. 05）.The level

关 键 词：甜菜碱 肝硬化 肝纤维化 内毒素 TOLL样受体 

分 类 号：R363[医药卫生—病理学] R965.2[医药卫生—基础医学]