楂曲平胃散抑制小鼠前脂肪细胞分化与防治肥胖机理的实验研究

【作者】 陈瑾；

【导师】 周训伦；

【作者基本信息】 成都中医药大学 ， 方剂学， 2006， 博士

【摘要】 目的：通过体外实验(细胞培养)研究楂曲平胃散抑制小鼠前脂肪细胞分化的作用及分子调控机制。通过体内实验(动物实验)研究楂曲平胃散预防小鼠肥胖和治疗肥胖小鼠的作用，并探讨其作用机制是否与其抑制前脂肪细胞分化的作用有关。 方法：(1)体外实验：加入楂曲平胃散处理原代培养小鼠前脂肪细胞，运用形态学方法观察细胞分化过程中的形态变化、运用油红0染色提取法检测细胞内的脂肪含量、运用酶组织化学提取法检测细胞内甘油磷酸脱氢酶(GPDH)的含量，同时运用反转录-聚合酶链式反应(RT-PCR)检测细胞分化相关基因C／EBP α、PPAR γ、ADD1／SREBP1、Pref-1 mRNA的表达。(2)体内实验：进行楂曲平胃散防治肥胖作用的研究，在实验过程中运用称量方法测定食量、体重和体脂重量，根据公式计算小鼠肥胖指数和脂肪系数；运用形态学方法观察脂肪细胞大小及肝、肾形态变化；运用生化方法测定血脂含量。 结果：(1)体外实验：楂曲平胃散作用于细胞后，从形态学角度细胞内脂肪颗粒出现的速度较慢且数量较少；细胞内脂肪含量和细胞分化标志酶—GPDH的含量均少于阴性对照组细胞：同时C／EBP α mRNA、PPAR γ mRNA、ADD1／SREBP1 mRNA的表达量减少，Pref-1 mRNA的表达量增多。4mg／ml和6 mg／ml剂量组效果明显。(2)体内实验：楂曲平胃散用药后，小鼠食量有一定减少，体重减轻、肥胖指数下降，体脂重量减轻、脂肪系数下降，血脂含量降低，脂肪细胞体积变小。高、中剂量组效果明显。同时，各组小鼠肝、肾形态学观察未见明显损伤性病变。更多还原

【Abstract】 Objective: To study the effect, mechanisms of Zhaqupingweisan solution on differentiation in primary mouse preadipocytes in vitro and prevention from to be obese , reducing weight in nutritional obesity mice in vivo.Methods: (1) We observed the morphological changes and post Zhaqupingweisan solution added into cultured primary mouse preadipocytes in vitro. We detected the quantity of intracytoplasmic lipids by oil red 0 staining and the quantity of GPDH in the cytoplasm of differentiated adipocytes. And the expression of C/EBPα 、 PPARγ、 ADD1/SREBP1 and Pref-1 mRNA relating to differentiation of adipocytes was detected by RT-PCR. (2) During the process of in vivo, we observed body weight, obese index, fat weight, weight ratio, adipocyte size and blood lipid.Results: (1) During the process of in vitro, the lipid droplet was less and smaller treated with Zhaqupingweisan solution as compared with the untreated control cells. The quantity of intracytoplasmic lipids and GPDH was less than the untreated control cells. RT-PCR showed that Zhaqupingweisan solution reduced the expression, of C/EBPα, PPARγ, ADD1/SREBP1 mRNA and increased the expression of Pref-1 mRNA. The groups of 4mg/ml and 6 mg/ml showed noticeable effects. (2) During the process of in vivo, body weight, obese index, fat weight, weight ratio, adipocyte size and blood lipid in the three Zhaqupingweisan solution groups were更多还原