非酒精性脂肪肝模型小鼠肝中脂肪酸组成分析

 【目的】通过检测非酒精性脂肪性肝病（NAFLD）模型小鼠肝中脂肪酸的含量，分析肝脏中脂肪酸组成与NAFLD之间的关系。【方法】采用连续6周饲喂高脂饲料并结合每周皮下注射2次CCl4的方法诱导C57BL/6J小鼠的NAFLD。小鼠肝脏中脂肪酸经乙醚提取和BF3/甲醇溶液衍生化处理后，采用气相色谱-质谱 (GC-MS) 选择离子方式进行检测。样品中脂肪酸通过检索质谱数据库和脂肪酸标准品的保留时间进行定性分析，采用外标法进行定量分析。【结果】病理学检查表明成功诱导C57BL/6J小鼠的NAFLD。与对照组相比，试验组小鼠肝中C14:0、C16:0、C18:0和C20:3含量显著升高 (P＜0.01)，C15:0、C18:1、C18:2和C18:3 (n-3)含量显著降低 (P＜0.01)，饱和脂肪酸（SFA）含量显著增加(P＜0.01)，其余类别的脂肪酸均显著降低 (P＜0.01)。n-6 和 n-3 多不饱和脂肪酸（PUFA）的产物/前体比值显著升高(P＜0.01)，n-6/n-3 PUFA [C20:4/(C20:5+C22:6)]的比值显著降低 (P＜0.01)。【结论】NAFLD小鼠肝脏中SFA含量增加、不饱和脂肪酸含量减少以及 n-6 PUFA与 n-3 PUFA代谢紊乱不仅是NAFLD产生的效果，也是促进NAFLD发展的因素。

Analysis of Fatty Acids Composition in the Liver of Mice with Non-alcoholic Fatty Liver Disease

【Objective】 The aim of this study was to determine the liver fatty acid composition of an experimental mouse model of NAFLD induced by high-fat feed and CCl4 and to assess the association between the change of liver fatty acids content and NAFLD. 【Method】C57BL/6J mice were given high-fat feed for six consecutive weeks to develop experimental NAFLD. Meanwhile, these mice were given subcutaneous injections of a 40% CCl4-vegetable oil mixture twice per week. Fatty acids in mice liver were extracted with diethyl and derived by BF3/methanol, then analyzed by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring. Qualitative analysis of fatty acids in mice liver was carried out according to the retrieval mass spectrometry database and the retention time of fatty acid standard. Quantitative analysis was determined by external standard method. 【Result】Pathological examination showed that NAFLD of C57BL/6J mice was successfully induced. Compared with the control group, high-fat feed and CCl4 led to significant increases in C14:0, C16:0, C18:0 and C20:3 (P＜0.01), and decreases in C15:0, C18:1, C18:2 and C18:3 (n-3) (P＜0.01) in the mouse liver. The treatment also led to an increase in saturated fatty acids (SFA) and decreases in other types fatty acids. An increase in the ratio of product/precursor n-6 (C20:4/C18:2) and n-3 [(C20:5+C22:6)/C18:3 (n-3)] and a decrease in the ratio of n-6/n-3 [C20:4/(C20:5+C22:6)] were also observed.【Conclusion】An increase of SFA, a decrease of unsaturated fatty acids (UFA), and metabolic disturbance of n-6 polyunsaturated fatty acids (PUFA) and n-3 PUFA in the liver of mice with NAFLD induced by high-fat feed and CCl4 were not only the results of NAFLD, but also the stimulating factors for the development of NAFLD.