基于PPARγ节点双向调控探寻葛根芩连汤改善脂肪胰岛素抵抗的分子网络机制

项目名称： 基于PPARγ节点双向调控探寻葛根芩连汤改善脂肪胰岛素抵抗的分子网络机制

项目编号： No.81460621

项目类型： 地区科学基金项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 涂珺

作者单位： 江西中医药大学

项目金额： 47万元

中文摘要： 2型糖尿病发病机制主要是胰岛素抵抗（IR）。中医认为IR与过食肥甘厚腻导致脾胃虚损相关，滋生痰湿与内热所诱发。葛根芩莲汤复方改善IR治疗糖尿病，临床疗效确切但干预机制不明确。研究表明小檗碱、黄芩苷、葛根素和甘草黄酮等影响与胰岛素增敏密切相关的PPARγ基因表达。这提示葛根芩连汤可能作用于代谢调控节点PPARγ来改善脂肪IR。本课题拟利用体内外IR模型，采用ELISA、Western blot、EMSA、荧光定量PCR和数字PCR等多种手段，重点研究究PPARγ调控的下游靶基因与脂肪分泌因子的分子调控网络。引入转录组芯片技术研究葛根芩莲汤干预糖尿病大鼠脂肪转录表达谱差异构建转录调控网络。结合代谢组学研究结果，系统阐明葛根芩连汤基于PPARγ双向调控通路改善脂肪IR的分子网络机制，为葛根芩连汤治疗糖尿病提供理论参考和科学依据，并为清热燥湿中药复方治疗代谢性疾病的多靶点网络作用机制研究提供新思路

中文关键词： 2型糖尿病；胰岛素抵抗；信号转导；脂肪细胞因子；过氧化物酶体增殖物激活受体γ

英文摘要： Diabetes and obesity have become a major threat to human health. The vast majority is type 2 diabetes mellitus (T2DM). The pathogenesis of T2DM is mainly insulin resistance （IR）and β-cell defects. Ge Gen Qin Lian Decoction (GGQLD) is now widely and effectively used to improve IR effect in clinical treatments of traditional Chinese medicines (TCM). It was reported that these active components of GGQLD such as berberine，bailcain,puerarin and glycyrrhizic acid influence the PPARγ expression level. Since the IR effect is found in adipose tissue in the earliest stage of T2DM, high expression of PPARγ from adipose tissue may be the target protein of GGQLD intervention. It is worthwhile for us to determine if PPARγ mediates lipid and glucose metabolic pathways upon GGQLD intervention by molecular biology, cell biology, biochemical and metabolic methods. Firstly, we will examine if mRNA and protein levels of PPARγ are significantly changed in adipose tissue isolated from in vivo and in vitro models; secondly, we will investigate the influence of PPARγ on the expression level of downstream target genes aP2/FABP4, LDL, CD36/ FATP-1, FASN，GK，GLUT4 and adipocytokines such as TNF-α, IL-6, leptin, resistin, visfatin and ADPN by ELISA ,Western blot and fluorescence quantitative RT-PCR. Furthermore, miRNAs regulation based on PPARγ mediated metabolic pathway from adipocyte cell will be studied in this proposal. mRNA and miRNA expression array will be employed in IR Rat model to obtain the expression profiles based on PPARγ regulation. This study will not only provide a theoretical basis for GGQLD treatment of T2DM but also provide important theoretical and realistic significance for the corresponding development of TCM preparation.

英文关键词： T2DM;Insulin Resistance (IR);Signal transduction;Adipocytokine;PPARγ