爪蟾卵母细胞固醇调控通路的建立及姜黄素的作用  

Establishing the SREBP Pathway in the Oocytes Xenopus Laevis and Stuyding the Effects of the Cucurmin

作　　者：沃立科[1] 范春雷[2] 沃兴德[2] 

机构地区：[1]浙江中医药大学附属第一医院,杭州310006 [2]浙江中医药大学生命科学学院

出　　处：《浙江中医药大学学报》2007年第3期288-290,共3页Journal of Zhejiang Chinese Medical University

基　　金：国家自然科学基金资助项目(No:30070932)~~

摘　　要：[目的]在爪蟾卵母细胞中建立人类SCAP/SREBP固醇调控系统,利用此系统研究姜黄素对SCAP/SREBP调控通路的作用。[方法]构建带有绿色荧光报告蛋白基因的PLXRN-4SRE-fpa质粒,将构建质粒和其他几种质粒分别提取混和成实验组质粒和对照组质粒,通过微注射的方法分别共注射到Ⅴ、Ⅵ爪蟾卵母细胞核内,与不同浓度的姜黄素共孵育3天,使用荧光定量PCR检测荧光表达量。[结果]相同浓度的姜黄素对注射含有固醇调节元件质粒的卵母细胞荧光表达量要明显高于不含相应元件的质粒。[结论]构建的质粒pLXRN-4SRE-fPA能在爪蟾卵母细胞中有效地应答SCAP、SREBP调控因子;通过影响SCAP/SREBP调控通路调节SRE-1下游基因的表达可能是姜黄素调脂作用的机理之一;利用核内共注射的方法在爪蟾卵母细胞中建立的SCAP/SREBP调控系统可作为SCAP配体类药物快速筛选和作用机制研究的细胞模型。[Aim] The expressed system of sterol regulation by SCAP/SREBP Pathway was established in the oocyte of the Xenopus laevis and was applied to investigate the effect of cucurmin on the SREBP pathway. [Methods] The plasmid of PLXRN-4SRE-fpa was restructed and injected into the nucleus of Xenopus Oocytes in phase Ⅴ or Ⅵ. The oocyte was incubated with different concentrations of the curcumin. After three days the fluorescence intensity on the membrane of the Oocytes was measured by Fluoroanalyzer[Results] The fluoresecence intensity on the membrane of the oocyte increased with increasing of the concentration of the curcumin. The trail group which have all regulation elements of SCAP/SREBP pathway can express more fluorscence protein than the control group. [Conclusion] The experiment provides that the curcumin can upgrade the expression of low density lipoprotein receptor by SCAP/SREBP regulation pathway. The model can be applied for quick screening of the scap ligand drugs and inverstigating the mechanism of the drugs.

关 键 词：爪蟾卵母细胞 姜黄素 SCAP/SREBP固醇调控通路 

分 类 号：R285.5[医药卫生—中药学]