摘要
目的:本研究探究了姜黄素促进白色脂肪细胞棕色化的作用及其潜在机制。方法:抽提小鼠皮下原代脂肪细胞,并诱导其分化成熟。予姜黄素(10、20、40μmol/L)孵育48 h,油红O染色观察脂肪细胞脂滴形态;使用线粒体绿色荧光探针(Mito Tracker Green)检测白色脂肪细胞内线粒体数量;运用线粒体超氧化物红色荧光探针(Mito SOX Red)测定细胞线粒体内活性氧(reactive oxygen species,ROS)水平;应用ATP试剂盒检测细胞内ATP含量;Real-time PCR检测棕色化相关标志性基因的表达水平;最后利用β3肾上腺素能受体(β3 adrenergic receptor,β3-AR)拮抗剂(SR59230A)进行阻断实验。结果:姜黄素能够减小脂肪细胞脂滴大小,增加脂肪细胞线粒体数量(P<0.001)。同时,低剂量姜黄素(10、20μmol/L)可减少线粒体ROS产生(P<0.01),提高细胞内ATP含量(P<0.05)。姜黄素干预能上调棕色脂肪标志性基因[β3-AR基因、PPARγ共激活因子-1α(PPARγcoactivator-1α,PGC1α)、解偶联蛋白1(uncoupling protein 1,Ucp1)]的mRNA表达水平(P<0.05)。而阻断β3-AR则能抑制姜黄素促进白色脂肪细胞棕色化的作用。结论:姜黄素通过β3-AR促进白色脂肪细胞线粒体生物合成并改善其功能,诱导白色脂肪细胞棕色化。
Objective:The aim of this study was to investigate the role of curcumin in promoting browning of white adipocytes and its underlying mechanisms through in vitro experiments.Methods:The subcutaneous primary adipocytes were extracted from mice and induced to differentiate into mature.Intervention with different concentrations of curcumin(10,20,40μmol/L)for 48 h.Lipid droplet morphology was observed by oil red O staining.Intracellular mitochondria were detected with mitochondrial green fluorescent probe(Mito Tracker Green).Mitochondrial superoxide indicator(Mito SOX Red)was used to determine the level of mitochondrial reactive oxygen species(ROS).Intracellular ATP content was detected by the ATP assay kit.The expression of the browning-related marker gene was detected by real-time PCR.Finally,the mechanism was explored by usingβ3 adrenergic receptor(β3-AR)antagonist(SR59230 A).Results:In mouse subcutaneous primary adipocytes,curcumin markedly alleviates intracellular lipid accumulation.And more mitochondria were observed after curcumin treatment(P<0.001).Otherwise,curcumin of low dose(10,20μmol/L)also reduced the production of ROS(P<0.01),but increased the intracellular ATP content(P<0.05).In addition,curcumin can up-regulate the mRNA expression of the brown fat marker genes[β3-AR,uncoupling protein 1(UCP1),PPARγcoactivator-1α(PGC1α)](P<0.05).However,β3-AR antagonist can antagonize the function of curcumin.Conclusion:Curcumin has the potential to promote mitochondrial biogenesis,improve their function and evoking white adipocyte browning through regulatingβ3-AR-dependent signaling pathway.
作者
朱晓蕾
杜苏苏
严琴慧
周炜
李晓南
ZHU Xiaolei;DU Susu;YAN Qinhui;ZHOU Wei;LI Xiaonan(Department of Child Health Care,the Affiliated Children's Hospital of Nanjing Medical University,Nanjing 210008,China)
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2020年第6期796-802,共7页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81773421)
江苏省重点研发计划(社会发展)重点病种的规范化诊疗(BE2015607)
江苏省科教强卫工程区学创新团队(CTXAD2017035)。
