载脂蛋白A5(ApoA5)经CIDEC负向调控人脂肪间充质干细胞(AMSC)成脂分化  

Effects of apolipoprotein A5(ApoA5) on inhibiting the progress of adipogenic differentiation of human AMSCs through CIDEC

作　　者：孔一 苏欣[2] 郑小燕[2] KONG Yi;SU Xin;ZHENG Xiao-yan(Department of Dermatology Department of Cardiovascular Medicine-,Central South University,Changsha 410012,Hunan Province,China;Department of Dermatology Department of Cardiovascular Medicine-,the Second Xiangya Hospital, Central South University,Changsha 410012,Hunan Province,China)

机构地区：[1]中南大学湘雅二医院皮肤与性病科,长沙410012 [2]中南大学湘雅二医院心血管内科,长沙410012

出　　处：《复旦学报（医学版）》2018年第6期775-780,共6页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金(81300205);中南大学研究生自主探索创新项目(2018zzts923)~~

摘　　要：目的探究载脂蛋白A5 (apolipoprotein A5,ApoA5)对人脂肪间充质干细胞(adipose mesenchymal stem cell,AMSC)成脂分化的影响及其机制。方法获取中南大学湘雅二医院腹部外科手术患者皮下脂肪组织,分离人AMSC,经成脂诱导剂诱导分化,以600、1 200 ng/mL ApoA5干预细胞,作为实验组;不加ApoA5干预的细胞作为对照组。定期收获细胞,观察ApoA5对细胞内脂滴油红O吸光度、三酰甘油(triglyceride,TG)含量和成脂标志物mRNA水平的影响。收获干预至14天的细胞,通过荧光抗体标记,利用激光共聚焦实验观察ApoA5与细胞死亡的DFF45样效应因子C (cell death-inducing DNA fragmentation factor 45-like effector C,CIDEC)是否存在共定位现象。定期收获细胞,分别应用real-time PCR和Western blot观察ApoA5对CIDEC、C/EBPβ的mRNA和蛋白质水平的影响。结果在同一促分化时间段内,ApoA5干预的细胞油红O吸光度、TG含量及aP2、FAS等成脂分化标志物mRNA表达水平下降。经过激光共聚焦分析发现,在AMSC成脂分化过程中,ApoA5与CIDEC存在共定位现象。在同一促分化时间段内,ApoA5可明显下调细胞内CIDEC、C/EBPβ的mRNA和蛋白质水平。结论在成脂分化过程中ApoA5与CIDEC共存,ApoA5通过下调CIDEC可抑制AMSCs成脂分化。Objective To verify whether apolipoprotein A5(ApoA5)could inhibit the adipogenic differentiation progress of human adipose mesenchymal stem cell(AMSC)and investigate the mechanism. Methods We isolated AMSCs by collagenase digestion from the adipocyte tissue of the patient underwent abdominal surgery.Then we used the adipogenic program to make AMSCs differentiate into mature adipocytes and incubated with 600 and 1 200 ng/mL ApoA5.The control group cultrued without ApoA5.We calculated the absorbance of oil Red O and the intracellular content of triglyceride(TG),and observed the gene expression levels of adipogenic differentiation markers.On the 14 th day after differentiation we used Alexa Fluor 488 fluorescent dye to label ApoA5 and then used the immunofluorescence to label sub-cellular organelles to observe whether ApoA5 and cell deathinducing DNA fragmentation factor 45-like effector C(CIDEC)have the co-localization phenomenon.We used real-time PCR and Western blot to detect the impact of ApoA5 on expression level of CIDEC and C/EBPβ. Results ApoA5 treatment could significantly decrease the amount of intracellular lipid droplets and TG concentration.During the same process of AMSCs differentiation,ApoA5 could dosedependently decrease the oil red O absorbance and the gene level of aP2 and FAS.Confocal microscopy analysis clearly showed that apoA5 co-localized with CIDEC in adipocytes.ApoA5 showed a concentration-dependently decreasing the mRNA and protein levels expression of CIDEC and C/EBPβ.Conclusions During the differentiation process of AMSCs,ApoA5 and CIDEC colocate.ApoA5 could inhibit the differentiation progress of AMSCs by down-regulating CIDEC.

关 键 词：载脂蛋白A5(ApoA5)$脂肪间充质干细胞(AMSC)细胞死亡的DFF45样效应因子C(CIDEC) 成脂分化 

分 类 号：Q26[生物学—细胞生物学]