OMEGA-3 FATTY ACID--CCL4 INDUCED LIVER DISEASE

本提案的目的是通过以下方式描绘分子机制 富含ω-3脂肪酸（ω-3FAs）的饮食对人体有害， 肝脏 因为鱼油引起的肝损伤可能会加重 同时使用另一种肝毒素，如酒精，两种药物 将被用来确定纤维化是否被增强。 具体 目的：1）确定单独使用3FAs的基因调控水平， 并与乙醇组合，引发肝纤维化; 2） 分析富含w3 FAs和乙醇的饮食对 类花生酸 3)为了确定w3 FAs和乙醇的作用， 4）为了确定施用对纤维化细胞因子合成的影响， w3 FAs和乙醇诱导肝损伤的分子机制 与CCl 4诱导的纤维化机制不同; 5）描述 特定生物制剂的抗纤维化作用。 方法：大鼠 将被喂食富含w3 FAs或富含饱和脂肪+/-乙醇的饮食， 六个月 将评价肝脏的胶原蛋白和其他细胞外 通过光学和电子显微镜观察基质基因表达，HPLC分析， 北方印迹杂交研究和核连续测定。 花生酸 肝组织的分析将通过HPLC和RIA完成。 级联 炎症和纤维化细胞因子将通过生物学方法进行研究。 稳态mRNA水平和基因转录的测定 rates. 凝胶阻滞试验和DNA酶I足迹法将用于 评估可能存在于CCl 4之间的核结合蛋白的差异 - 诱导的纤维化和损伤。 生物制剂如PGE 2、γ-干扰素和中和 TGF β或TNF α的抗体将用于干预 纤维化过程 肝细胞、Ito细胞和 枯否细胞也将评估w3 FAs和乙醇对基质的影响。 形成、类花生酸代谢和细胞因子产生。 健康 相关性：假设w3 FAs可能在肝脏中发挥作用， 纤维化是具有重要治疗意义的新概念;如果 被证明是正确的，这一理论将提出严重的问题， 滥用w3 fas。

The objective of this proposal is to delineate the molecular mechanisms by which a diet rich in omega 3 fatty acids (w 3FAs) is injurious to the liver. Because the hepatic injury induced by fish oil may be exacerbated by the concomitant use of another hepatotoxin, such as alcohol, both agents will be employed to determine whether fibrogenesis is enhanced. Specific Aims: 1) to determine the level of gene regulation by which w 3FAs alone, and in combination with ethanol, initiate hepatic fibrogenesis; 2) to analyze the actions of a diet rich in w3FAs and ethanol on the formation of eicosanoids. 3) to ascertain the effects of w3FAs and ethanol administration on fibrogenic cytokine synthesis; 4) To determine whether the molecular mechanisms by which w3FAs and ethanol induce liver injury differ from the mechanisms of CCl4-induced fibrosis; and 5) to delineate the antifibrogenic effects of specific biological agents. Methods: Rats will be fed a diet rich in w3FAs or rich in saturated fats +/- ethanol for six months. Livers will be evaluated for collagen and other extracellular matrix gene expression by light and electron microscopy, HPLC analysis, Northern blot hybridization studies, and nuclear run-on assays. Eicosanoid analysis of liver tissue will be accomplished by HPLC and RIA. The cascade of inflammatory and fibrogenic cytokines will be investigated by biological assays and determination of steady state mRNA levels and gene transcription rates. The gel retardation assay and DNase I footprinting will be used to assess differences in nuclear-binding proteins that may exist between CCl4 - induced fibrosis and injury induced by w3FAs and ethanol administration. Biological agents such as PGE2, gamma-interferon, and neutralizing antibodies to TGF beta or TNF alpha will be used to intervene in the fibrogenic process. In vitro studies of hepatocytes, Ito cells, and Kupffer cells will also evaluate the effects of w3FAs and ethanol on matrix formation, eicosanoid metabolism, and cytokine production. Health relatedness: The hypothesis that w3FAs may play a role in hepatic fibrogenesis is a novel concept with important therapeutic implications; if shown to be true, this theory would raise serious questions about the indiscriminate use of w3FAs.