EGCG调控3T3-L1脂肪细胞向棕色脂肪分化的作用与机制研究  

Role and mechanism of EGCG in regulating the differentiation of 3T3-L1 adipocytes to brown fat

作　　者：郭铖洁 侯森 吕世文[3] GUO Chengjie;HOU Sen;LV Shriwen(Jinhuxr Maerral&Child Health Care Hospital Jinhua 321000,Zhejiang,China)

机构地区：[1]金华市妇幼保健院,浙江省金华321000 [2]济南护理职业学院 [3]金华市中心医院

出　　处：《现代实用医学》2024年第7期848-853,共6页Modern Practical Medicine

基　　金：金华市科技计划项目(2022-4-188)。

摘　　要：目的探讨表没食子儿茶素没食子酸酯(EGCG)调控3T3-L1脂肪细胞向棕色脂肪分化的作用与机制。方法培养3T3-L1脂肪细胞并诱导其分化,通过MTT法检测不同EGCG浓度对细胞增殖的影响,选取浓度为10、50、100μmol/L的EGCG处理3T3-L1脂肪细胞,空白对照组不做处理。采用AMPK抑制剂Compound C和AMPK激活剂AICAR分别处理3T3-L1脂肪细胞。油红O染色观察EGCG对3T3-L1细胞分化的影响;qRT-PCR法和Western blot法检测3T3-L1脂肪细胞分化及褐化相关因子及蛋白的表达水平;用Mito-tracker-Green探针检测线粒体荧光。结果随EGCG浓度的增加,3T3-L1细胞的脂滴减少,且呈一定浓度依赖性。加入AMPK激活剂AICAR可进一步减少脂滴,而加入AMPK抑制剂Compound C可逆转脂滴减少情况。随EGCG浓度的升高,PPARr、FASN mRNA及蛋白的表达水平均降低,UCP1、PGC-1α、Nrf1、Tfam、ATGL、HSL、AMPK mRNA及蛋白表达水平均升高(均P<0.05),加入AMPK激活剂AICAR可进一步加重此趋势,而加入AMPK抑制剂Compound C可逆转此趋势。荧光显微镜观察发现EGCG处理后荧光强度相对于对照组显著增强。结论EGCG通过调控3T3-L1脂肪细胞AMPKα/PGC-1α信号途径促进脂肪细胞棕色化和线粒体生物合成。Objective To investigate the role and mechanism of EGCG on the differentiation of 3T3-L1 adipocytes into brown adipocytes.Methods 3T3-L1 adipocytes were grown and divided,and the influence of cell proliferation was detected by MTT assay with different concentrations of EGCG.3T3-L1 adipocytes were treated with different concentrations of 10,50,and 100 μmol/L of EGCG,and no treatment was done in the blank control group.AMPK inhibitor Compound C and AMPK activator AICAR were used to treat 3T3-L1 adipocytes respectively.The influence of EGCG was observed on the polarizing effect of 3T3-L1 cells with oil red O staining;the expression levels of factors related to 3T3-L1 adipocyte differentiation and browning were detected by qRT-PCR;the expression levels of proteins related to 3T3-L1 adipocyte differentiation and browning were detected by western blotting analysis;and the mitochondrial fluorescence was detected by Mitochondrial Green probe.Results The lipid droplets of 3T3-L1 cells decreased with increasing EGCG levels in a concentration-dependent manner.Addition of the AMPK activator AICAR further reduced lipid droplets,whereas addition of the AMPK inhibitor Compound C reversed the lipid droplet reduction.With the increase of EGCG concentration,the expression levels of PPAR,FASNN mRNA and protein were decreased,and the expression levels of UCP1,PGC-1α,Nrf1,Tfam,ATGL,HSL,AMPK mRNA and protein were increased(all<0.05),and this trend was further aggravated by the addition of the AMPK activator,AICAR,which could be further aggravated by the addition of the AMPK inhibitor Compound C.Fluorescence microscopy observation revealed that the fluorescence intensity was significantly enhanced compared to the control group after EGCG treatment.Conclusions EGCG promotes adipocyte browning and mitochondrial biosynthesis by regulating the AMPKα/PGC-1α signaling pathway in 3T3-L1 adipocytes.

关 键 词：表没食子儿茶素没食子酸酯 3T3-L1脂肪细胞 分化 白色脂肪棕色化 

分 类 号：R589.2[医药卫生—内分泌]