摘要
目的 探索鸢尾黄素对前脂肪细胞分化的影响及机制.方法 CCK8 法检测鸢尾黄素对 3T3-L1 细胞存活率的影响.细胞接触抑制 2d(分化第0 天),加入诱导剂和不同浓度鸢尾黄素(0、10、20、40、60 μmol/L)处理细胞,对照组不加诱导剂.分化第 9 天,油红O染色观察脂滴形成,生化试剂盒检测TG、NEFA水平,Western blot法检测PPARγ、C/EBPα、perilipin-1、ADFP、AMPKα、p-AMPKα 蛋白表达,RT-qPCR 法检测 Adiponectin、FABP4、FAS、Acly mRNA表达.结果 鸢尾黄素浓度≤60 μmol/L时不影响细胞存活率(P＞0.05).与诱导分化组比较,鸢尾黄素给药后,细胞内脂滴形成减少,TG、NEFA水平降低(P＜0.01);C/EBPα、PPARγ、perilipin-1、ADFP蛋白表达降低(P＜0.01),AMPKα、p-AMPKα 蛋白表达升高(P＜0.01);Adiponectin、FABP4、FAS、Acly mRNA 表达降低(P＜0.01).结论 鸢尾黄素具有抑制前脂肪细胞分化为成熟脂肪细胞的作用,其机制可能与C/EBPα、PPARγ、AMPKα和p-AMPKα的调控有关.
Abstract
AIM To investigate the effects of tectorigenin on preadipocyte differentiation and the possible mechanism.METHODS CCK8 method was used to detect the effects of tectorigenin on 3T3-L1 cell viability.After 2 days contact inhibition(Day 0 of differentiation),the cells were exposed to inducer and tectorigenin of different concentrations(0,10,20,40,60 μmol/L),in contrast to those of the control group with no inducer use.On the 9th day of differentiation,the cells had their lipid droplets observed by oil red O staining;their levels of TG and NEFA detected by biochemical kit;their protein expressions of PPARγ,C/EBPα,perilipin-1,ADFP,AMPKα and p-AMPKα detected by Western blot;and their mRNA expressions of Adiponectin,FABP4,FAS and Acly detected by RT-qPCR.RESULTS Tectorigenin concentration of 60 μmol/L or lower levels left no impact upon the cell viability(P＞0.05).Compared with the model group with induced differentiation,the groups intervened with tectorigenin administration displayed decreased formation of lipid droplets;lower levels of TG and NEFA(P＜0.01);decreased protein expressions of C/EBPα,PPAR γ,perilipin-1 and ADFP(P＜0.01);increased protein expressions of AMPKα and p-AMPKα(P＜0.01);and decreased mRNA expressions of Adiponectin,FABP4,FAS and Acly(P＜0.01).CONCLUSION Tectorigenin inhibits preadipocyte differentiation into mature adipocytes,which may be related to its efficacy in the regulation of C/EBPα,PPARγ,AMPKα and p-AMPKα.
基金项目
上海市医学重点专科项目(ZK2019B16)
上海市卫生健康委科研项目(20204Y0154)
上海市卫生健康委科研项目(202240309)
上海市普陀区临床特色专科项目(2020tszk01)
上海市普陀区卫生健康系统科技创新项目(ptkwws202003)
维普
万方数据