过氧化物酶体增殖物激活受体γ2在脂肪干细胞脂向分化过程中的表达  

Expression Profiling of Peroxisome Proliferation Activate Receptor γ2 and Phosphorylation During Adipogenic Differentiation of Adipose-derived Stem Cells

作　　者：孙钦策[1] 肖金刚[2,3] 杨晓娟[2] 田卫东[2,4] 

机构地区：[1]四川大学生命科学学院生物化学与分子生物学系,成都610064 [2]四川大学口腔疾病研究国家重点实验室,成都610041 [3]泸州医学院附属口腔医院口腔颌面外科,泸州646000 [4]四川大学华西口腔医院颅颌面外科,成都610041

出　　处：《四川大学学报（医学版）》2011年第1期10-14,共5页Journal of Sichuan University(Medical Sciences)

基　　金：教育部博士点基金(20070610064);国家自然科学基金(批准号30973348)资助

摘　　要：目的观察转录因子过氧化物酶体增殖物激活受体γ2(peroxisome proliferation activate receptorγ2,PPARγ2)及其磷酸化形式在脂肪干细胞脂向分化过程中的表达情况,为研究脂肪发育的分子机制提供新线索。方法组织块贴壁法培养脂肪干细胞,采用条件培养基(含有α-改良型、10%胎牛血清、10-6mol/L地塞米松磷酸钠、10-5mol/L胰岛素、0.2mmol/L吲哚美辛和0.5mmol/L3-异丁基-1-甲基黄嘌呤)对第3代细胞进行脂向分化诱导,RT-PCR、Western blot、免疫荧光技术检测在诱导的第0d、1d、3d、5d、7d、9dPPARγ2非磷酸化及其磷酸化形式的表达情况。结果随着细胞脂向分化时间的延长,PPARγ2非磷酸化及其磷酸化形式的基因和蛋白表达量都明显增加。结论 PPARγ2是脂肪分化的关键调节者,推测PPARγ2可能通过自身的磷酸化来促进脂肪干细胞的脂向分化。Objective To investigate PPARγ2 expression and phosphorylation involved in adipogenic differentiations of Adipose-derived Stem Cells（ASCs） with an aim to reveal the possible mechanisms of adipose development.Methods ASCs were obtained from the inguinal fat pads of GFP（green fluorescent protein） mice.The primary cells were cultured in histiocytic attachment cultivation.Adipogenic differentiation of the third generation of ASCs was induced with the conditional medium（α-MEM medium supplemented with 100 mL/L fetal bovine serum,10-6 mol/L dexamethasone,10-5 mol/L insulin,0.2 mmol/L indomethacin,and 0.5 mmol/L 3-isobutyl-1-methylxanthine）.The levels of PPARγ2 mRNA and the PPARγ2 expression and phosphorylation at day 0,1,3,5,7,and 9 of induction were detected using RT-PCR,fluorescence-immunocytochemistry and western blot,respectively.Results The expression of PPARγ2 and phosphorylation were significantly up-regulated during adipogenic differentiation of ASCs.Conclusion PPARγ2 is the key regulator of adipose development.The regulation of PPARγ2 phosphorylation may promote adipogenic differentiation of ASCs.

关 键 词：脂肪干细胞 过氧化物酶体增殖物激活受体γ2 磷酸化 脂肪分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]