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目的探讨CYP1B1对高脂膳食诱导的成年小鼠脂肪代谢的作用。方法 CYP1B1基因敲除(KO)和野生型(WT)雄性成年C57/BL小鼠(6 w龄)各16只,给予低脂(LFD,30%)、高脂肪(HFD,60%)饲料共6 w。小鼠处死后取血清、附睾脂肪和肝脏组织检测相应的生化和分子生物学指标。结果 6 w高脂膳食后,KO小鼠能量摄入总量稍高于WT小鼠,但其体重增量和附睾脂肪组织重量均显著低于WT小鼠;WT小鼠脂肪细胞直径明显大于KO小鼠,且血糖、血清及肝脏组织中甘油三酯(TG)水平亦明显高于KO小鼠;肝脏组织RT-PCR结果显示,CYP1B1基因敲除后,启动脂肪形成的核因子及脂肪合成相关基因如CD36、SREBP1c、SCD1等表达下降,而调控脂肪氧化分解的基因如CPT-1α,UCP-2表达显著上升;蛋白印迹结果显示,CYP1B1基因敲除增强腺苷-磷酸激酶(AMPK)的磷酸化。结论 CYP1B1基因敲除对成年小鼠营养性肥胖的保护作用可能与AMPK磷酸化增强并调控肝脏中脂肪代谢相关基因的表达有关。
Objective To investigate the effect of CYP1B1 on fat metabolism induced by high-fat diet in adult mice. Methods Sixteen male C57 / BL mice (6 weeks old) with CYP1B1 knockout (KO) and wild type (WT) were given low fat diet (LFD 30%) and high fat diet 6 w. After sacrifice, the mice were sacrificed and serum, epididymal fat and liver tissues were tested for biochemical and molecular biological indicators. Results After 6 w high-fat diet, the total energy intake of KO mice was slightly higher than that of WT mice, but the body weight gain and the weight of epididymal adipose tissue were significantly lower than those of WT mice. The diameter of adipocytes in WT mice was significantly greater than KO The levels of triglyceride (TG) in blood glucose, serum and liver tissue of mice were also significantly higher than that of KO mice. The RT-PCR results of liver tissue showed that CYP1B1 gene knockdown activated the formation of fat-related nuclear factor and fat synthesis The expression of genes such as CD36, SREBP1c and SCD1 decreased while the expression of genes such as CPT-1α and UCP-2 that regulated lipoxygenase degradation increased significantly. Western blotting showed that CYP1B1 knockdown enhanced phosphorylation of adenosine-phosphate kinase (AMPK) The Conclusion The protective effect of CYP1B1 knockout on nutritional obesity in adult mice may be related to the enhancement of phosphorylation of AMPK and regulation of the gene expression of fat metabolism in liver.