刺芒柄花素通过促进解偶联蛋白1(UCP1)表达刺激棕色脂肪细胞产热  

Formononetin stimulates thermogenesis of brown adipocytes by promoting the expression of uncoupling protein 1

作　　者：陶金[1] 陶磊 黄文华[2] TAO Jin;TAO Lei;HUANG Wenhua(General Hospital of Ningxia Medical University,Yinchuan 750004;Ningxia Hui Autonomous Region People’s Hospital,Yinchuan 750001,China)

机构地区：[1]宁夏医科大学总医院,宁夏银川750004 [2]宁夏回族自治区人民医院,宁夏银川750001

出　　处：《细胞与分子免疫学杂志》2020年第3期242-247,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：宁夏医科大学校级课题(XM2018104)。

摘　　要：目的探讨刺芒柄花素(formononetin)通过解偶联蛋白1(UCP1)调控棕色脂肪细胞产热的机制。方法分离来源于C57BL/6J小鼠的脂肪组织的前体细胞,体外诱导为成熟的棕色脂肪细胞,利用实时定量PCR检测脂肪细胞标志基因脂肪酸结合蛋白4(FABP4)、脂连蛋白(Adiponectin)的表达确定其脂肪细胞的特性;利用实时定量PCR及Western blot法检测formononetin是否促进棕色脂肪细胞产热基因过氧化物酶体增殖物激活受体γ共刺激因子1α(PGC-1α)、碘甲腺原氨酸脱碘酶2(Dio2)、过氧化物酶体增殖剂激活受体γ(PPARγ)及解偶联蛋白1(UCP1)的表达;分离来自UCP1敲除小鼠的棕色脂肪前体细胞,并诱导分化至成熟的脂肪细胞并检测分化效率。上述细胞的对照为来自野生型C57BL/6J小鼠分化成熟脂肪细胞;利用XF24细胞能量代谢仪检测formononetin对野生型及UCP1敲除棕色脂肪细胞呼吸的影响。结果野生型和UCP1敲除的棕色脂肪前体细胞均能诱导分化至成熟脂肪细胞;Formononetin能促进野生型棕色脂肪细胞产热基因PGC-1α、Dio2、PPARγ及UCP1的表达,也能促进UCP1敲除棕色脂肪细胞中UCP1上游产热基因PGC-1α、Dio2的表达;UCP1的缺失抑制formononetin诱导棕色脂肪细胞呼吸的氧耗量。结论Formononetin通过促进UCP1的表达从而激活棕色脂肪细胞的产热及氧耗量。Objective To investigate the mechanism of formononetin regulating the heat production of brown adipocytes via decoupling protein 1(UCP1).Methods The brown preadipocytes was isolated from wild-type(WT)C57BL/6J mice and differentiated into mature fat cells in vitro.Moreover,the mRNA levels of fatty acid binding protein 4(FABP4)and adiponectin were detected by real-time quantitative PCR(RT-qPCR).To confirm formononetin could induce the expression of thermogenic genes,we first prepared WT mature brown adipocytes and treated them with DMSO and formononetin separately.The mRNA and protein levels of thermogenic genes,such as peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α),peroxisome proliferators-activated receptorγ(PPARγ),UCP1 and iodothyronine deiodinase 2(Dio2),were detected by RT-qPCR and Western blot analysis.To investigate the role of UCP1 in mediating differentiation of brown preadipocytes,Fabp4 and adiponectin mRNA levels were analyzed by RT-qPCR in WT and UCP1 mutation differentiated brown adipocytes.To determine cellular oxygen consumption,isolated WT and UCP1 mutation brown preadipocytes were plated in an XF24-well microplate and differentiated into mature brown adipocytes treated with formononetin or DMSO,followed by oxygen consumption rate(OCR)measurement using XF24 analyser.Results Both WT and UCP1 KO brown preadipocytes could be differentiated into adipocyte.The expression of thermogenic genes,including PGC-1α,Dio2,PPARγand UCP1,induced by formononetin was similar in UCP1 KO adipocytes and WT cells.But the ability of formononetin to increase cellular respiration was inhibited in Ucp1 KO cells.Conclusion Formononetin mediated stimulation of thermogenesis and oxygen consumption via UCP1 in brown fat cells.

关 键 词：刺芒柄花素(formononetin) 棕色脂肪细胞 解偶联蛋白1(UCP1) 产热 

分 类 号：R285.5[医药卫生—中药学] R392-33[医药卫生—中医学]