基于基因芯片技术的五味子素B诱导高甘油三酯血症小鼠模型作用机制研究＊

目的 探讨五味子素B（Schisandrin B,Sch B）诱导高脂血症小鼠模型的基因（肝脏）表达谱的变化及可能的机制。方法 ICR小鼠,雄性,分为7组,每组10只：正常饮食（normal diet,ND）组（2组）、ND/Sch B（0.5,2 g·kg^-1）组、高脂/糖饮食（high fat/fructose diet,HFFD）组、HFFD/Sch B（0.5,2 g·kg^-1）组。小鼠用Sch B（橄榄油配制）灌胃,48 h后用生化法检测血清和肝脏甘油三酯（TG）和总胆固醇（TC）水平,血清高密度脂蛋白胆固醇（HDL）和低密度脂蛋白胆固醇（LDL）水平;ELISA法检测血清极低密度脂蛋白胆固醇（VLDL）、载脂蛋白A1（Apo A1）、Apo E、Apo B48、Apo B100、Apo CⅡ和Apo CⅢ水平;HE染色观察肝脏病理学变化;油红O染色观察肝脏脂质沉积;通过基因芯片技术分析各组小鼠肝脏中全基因表达谱的变化及所涉及的通路。结果 ND/Sch B组小鼠血清/肝脏TG（升高266%/352.57%）和TC水平显著升高（升高36.54%/25.70%）,血清HDL和LDL水平显著升高（分别升高29.96%和30.68%）,但VLDL、Apo E、Apo B100、Apo CⅡ和Apo CⅢ显著降低（分别降低15.98%、28.90%、20.76%、20.53%和17.82%）;肝脏脂质沉积并出现病理学损伤;与HFFD组比,HFFD/Sch B组血清TG水平、肝脏TG和TC水平升高,血清HDL、LDL、VLDL、Apo E、Apo B100和Apo CⅡ水平显著降低;ND/Sch B组与ND组比肝脏有1016个基因差异表达,其中上调基因722个,下调基因294个,HFFD/Sch B组与HFFD组比肝脏有1162个基因差异表达,其中上调基因671个,下调基因491个。而HFFD组与ND组比肝脏有2070个基因差异表达,其中上调基因1289个,下调基因781个。Pathway分析结果显示Sch B诱导的高脂血症与11条通路的改变有关。结论 Sch B诱导高脂血症的机制与肝脏调节脂代谢多条通路、载脂蛋白及脂蛋白代谢密切相关。

Mechanism Investigation on the Hypertriglyceridemia Mouse Model-induced by Schisandrin B Based on Gene Chip Technology

Objective To investigate the changes of gene expression profile involved in Schisandrin B (Sch B)-induced hyperlipidemia and the underlying mechanism(s) of hyperlipidemia in mice.Methods Male ICR mice were randomly divided into various groups: (1) mice fed with normal diet (ND); (2) mice fed with ND and orally treated with Sch B at a bolus dose of 0.5 or 2 g·kg-1 (ND/Sch B); (3) mice fed with high fat/fructose diet (HFFD, fat, 10%; fructose, 10%; w/w); (4) mice fed with HFFD and orally treated with Sch B at a bolus dose of 0.5 or 2 g·kg-1 (HFFD/Sch B). Forty-eight hours after the Sch B treatment, blood samples were obtained from the orbital vein. Serum levels of triglyceride (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were analyzed by biochemical methods. Levels of very-low-density lipoprotein (VLDL), apolipoprotein A1 (Apo A1), Apo E, Apo B48, Apo B100, Apo CII, and Apo CIII were determined by ELISA methods. Liver tissues were stained by HE and histological changes were examined under the microscope. Lipid deposition in liver tissue was viewed after staining with oil red O. Gene-sequencing technology was applied to study the alterations of gene expression profile and the relevant signaling pathways.Results Serum and liver levels of TG and TC, as well as HDL and LDL levels, were significantly increased, but VLDL, Apo E, Apo B100, Apo CII, and Apo CIII levels showed significant decreases in ND/Sch B group when compared with those in ND group. Lipid deposition and pathological damages in the liver were also observed. HFFD/Sch B group showed significant increases in serum/liver TG and liver TC levels, but serum levels of HDL, LDL, VLDL, Apo E, Apo B100, and Apo CII were markedly decreased when compared with the HFFD group. When compared with the ND group, mice in the ND/Sch B group showed differential expression of 1016 genes, including 722 up-regulated genes and 294 down-regulated genes, in the liver. HFFP/Sch B group exhibited differential expression of 1162 genes, including 671 up-regulated genes and 491 down-regulated genes, when compared with the HFFD group. As compared to the ND group, HFFD-fed mice showed differential expression of 2070 genes, including 1289 up-regulated genes and 781 down-regulated genes. The results of pathway analysis indicated that the changes of 11 pathways were related to hyperlipidemia induced by Sch B.Conclusions The mechanism of Sch B-induced hyperlipidemia is closely related to the regulation of multiple pathways of lipid metabolism, apolipoprotein, and lipoprotein metabolism in the liver.