山茱萸环烯醚萜苷类提取物对3T3-L1脂肪细胞线粒体氧消耗及相关基因表达的影响  

Effects of Iridoid Glycoside Extract from Corni Fructus Extract on Mitochondrial Oxygen Consumption and Related Genes Expression in 3T3-L1 Mature Adipocytes

作　　者：赵丹丹[1] 李小可[1] 于娜[1] 穆倩倩 张毅[1] 方心[1] 莫芳芳[1] 高思华[1] 

机构地区：[1]北京中医药大学糖尿病研究中心,北京100029

出　　处：《中国实验方剂学杂志》2016年第11期122-126,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家国际科技合作项目(2011DFA30920);国家"重大新药创制"科技重大专项(2012ZX09103201-005)

摘　　要：目的:观察山茱萸环烯醚萜苷类提取物对小鼠胚胎成纤维细胞(3T3-L1)成熟脂肪细胞有氧呼吸氧消耗速率及相关基因表达的影响,分析可能分子机制。方法:以3T3-L1脂肪细胞为研究对象,Seahorse细胞外流量分析方法测定0.04,0.2,1 g·L^(-1)质量浓度山茱萸提取物干预2 d后对脂肪细胞氧消耗速率的影响,另设空白组。逆转录-聚合酶链式反应(RTPCR)法检测干预2 d后脂肪细胞过氧化物酶体增殖物激活受体-γ辅激活子-1α(PGC-1α),沉默信息调节因子2相关酶1(SIRT1),腺苷酸活化蛋白激酶(AMPK),线粒体脱偶连蛋白1抗体(UCP-1),PR结构域的蛋白16(PRDM16)和过氧化物酶体增殖物活化受体γ(PPARγ)mRNA表达水平。结果:与空白组比较,0.2,1 g·L^(-1)山茱萸提取物干预成熟脂肪细胞2 d后可增加ATP生成、基础呼吸和最大呼吸能力,1 g·L^(-1)提取物可使储备呼吸能力增强,0.04 g·L^(-1)提取物则使质子漏氧消耗速率明显升高;各质量浓度提取物干预2 d不同程度明显上调PGC-1α,AMPK,SIRT1 mRNA水平,明显下调PPARγmRNA表达,均具有明显统计学差异(P<0.05,P<0.01),对UCP-1,PRDM16基因表达没有明显影响。结论:山茱萸环烯醚萜总苷对成熟脂肪细胞有氧呼吸具有促进作用,增加ATP生成,细胞不具备米色化特征,其作用可能与AMPK,SIRT1,PGC-1α,PPARγ多个因子的调节相关。Objective： To observe the effects of iridoid glycoside extract from Corni Fructus（ ECF） on mitochondrial oxygen consumption and related genes expression in 3T3-L1 mature adipocytes, and analyze the possible molecular mechanism. Method： With 3T3-L1 adipocytes as the study objects,Seahorse XF Cell Mito Stress Test was used to detect the effect of ECF（ at the concentrations of 0. 04,0. 2,1 g·L^-1） for 2 d on the oxygen consumption rate in adipocytes,and blank group was additionally set up. Peroxisome proliferators-activated receptor-γ coactivator-1α（ PGC-1α）, silent information regulator 2 related enzyme 1（ SIRT1）, adenosine monophosphate activated protein kinase（ AMPK）,mitochondrial uncoupling protein 1 antibodies（ UCP-1）,PR domain proteins 16（ PRDM16） and peroxisome proliferators-activated receptor γ（ PPARγ） mRNA expressions were detected by reverse transcription-polymerase chain reaction（ RT-PCR）. Result： As compared with the blank group,2 days＇ intervention of 0. 2,1 g·L^-1ECF in mature 3T3-L1 adipocytes enhanced ATP production,basal respiration and maximal respiration; 1 g·L^-1ECF intervention enhanced the spare respiration capacity; 0. 04 g·L^-1ECF significantly increased the oxygen consumption rate of proton leak. ECF intervention for 2 days at various concentrations significantly up-regulated PGC-1α, AMPK and SIRT1 mRNA levels, and significantly downregulated PPARγ mRNA expression level,with statistically significant differences（ P〈0. 05,P〈0. 01）,but they showed no significant effects on UCP-1 and PRDM16 mRNA expression levels. Conclusion： In mature adipocytes,ECO could enhance the aerobic respiration and increase ATP production, and the adipocytes have no beiging characteristics,so these effects might be related to regulations of AMPK,SIRT1,PGC-1α and PPARγ levels.

关 键 词：山茱萸提取物 环烯醚萜苷 成熟脂肪细胞 氧消耗速率 质子漏 

分 类 号：R285.5[医药卫生—中药学]