摘要
目的探讨过氧化物酶体增殖物激活受体γ(PPARγ)基因转染对骨髓间充质干细胞(MSC)向脂肪细胞早期分化的调控作用。方法原代培养新西兰大白兔MSC,应用脂质体转染法将pEGFP-N1-PPARγ表达载体转入MSC中,G418筛选,成脂诱导剂诱导向脂肪细胞分化,分成不诱导组(A组)、空转染诱导组(B组)及转染诱导组(C组),采用RT-PCR方法检测PPARγ、脂蛋白脂酶(LPL)mRNA表达,免疫细胞化学染色检测PPARγ、LPL蛋白表达,油红O染色法行脂肪细胞计数和定量。结果A组细胞内没有脂滴出现,C组成脂率和脂肪含量明显高于B组(P<0.05);A组PPARγ、LPL mRNA及蛋白均不表达,C组PPARγ、LPL mRNA及蛋白表达水平显著高于B组(P<0.01)。结论PPARγ基因转染可增强MSC向脂肪细胞早期分化的能力,缩短分化进程,提高分化效率。
Objective To explore the modulation of genetic transfection of peroxisome proliferator-activated receptor γ （PPARγ） on early differentiation of rabbit marrow stroma cells into adipocytes. Methods The MSCs of the New Zealand rabbit were cultured which have been put in the expression vector of pEGFP-N1-PPARγ, with the method of lipoplast, then alternated by G418. Adipocytic differentiation was induced by inducers and was divided three group： uninduced group （Group A） ,untransfected group（ Group B） and transfected group（ Group C）. RT-PCR was employed to detect the expression levels of PPARγ, and lipoprotein lipase （LPL） mRNA. Immuncellulochemica was used to observe the expression of PPARγ and LPL protein. The counting and quantitation of adipose cell were examined by cell morphology and Oil-Red O staining measurement. Results There was no adipocytes in Group A. The rate and the counting of adipocytes in Group C were markedly higher than those in Group B. There were no expression of PPARγ and LPL. The expression levels of PPARγ and LPL Group C were increased significantly compared with Group B （P 〈 0.01 ）. Conclusions PPAR5, genetic transfection can reinforce the ability that MSCs early differentiate into adipocytes, decrease the differentiated time and increase the differentiated efficiency.
出处
《山东医药》
CAS
北大核心
2008年第28期16-18,共3页
Shandong Medical Journal
基金
湖南省卫生厅科研基金项目资助(B2006-116)
