摘要
目的:探讨过氧化物酶体增殖物激活受体γ(PPARγ)基因表达在骨髓间充质干细胞(MSC)向脂肪细胞早期分化中的调控作用及对脂肪分化相关蛋白(ADRP)表达的影响.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,RT-PCR检测PPARγ,ADRPmR-NA表达,Western Blot检测ADRP蛋白表达,细胞形态学、油红O染色法行脂肪细胞计数和定量.结果:未分化的MSC中PPARγ,ADRP mRNA不表达,经pEGFP-N1-PPARγ转染后,调控成脂肪细胞方向分化的转录因子和早期标志PPARγ mRNA(0.87±0.08vs0.28±0.01,P<0.01),ADRP mRNA(0.52±0.03vs0.21±0.02,P<0.01)表达增强;MSC向成脂肪细胞方向分化,成脂率及脂质量提高,分别为(78.5±4.2)%vs(51.4±3.0)%和(0.46±0.05)vs(0.21±0.02),P<0.05;分化成脂的细胞ADRP蛋白表达呈阳性,转染组较空转染组表达增强.结论:PPARγ在MSC向脂肪细胞分化的早期中起重要的正向调节作用,并促进ADRP基因和蛋白的表达;PPARγ基因过表达可增强MSC向脂肪细胞分化的能力,缩短分化进程,提高分化效率,可能与增强ADRP的表达有关.
AIM： To study the regulatory role of peroxisome proliferator-activated receptor γ（PPARγ） in the early differentiation of marrow stromal cells （MSCs） into adipocytes and its effect on the expression of adipose differentiation related protein （ADRP）. METHODS ：Primary New Zealand rabbit MSCs were collected. The expression vector pEGFP-N1-PPARγ was constructed and transfected into MSCs by liposome. Transfected MSCs were screened by G418. Adipocytic differentiation was induced by inducers. RT-PCR was used to measure the expressions of PPART,ADRPmRNA. Protein expression was measured by Westem Blot. Adipocytes were calculated under microscope and lipid droplets in the cells were stained with Oil-Red O. RESULTS： The undifferentiated MSCs did not express PPARγ or ADRPmRNA. After transfected with pEGFP-N1-PPARγ, MSCs differentiated into adipocytes, and the expressions of PPART, ADRP mRNA were increased significantly. The adipogenic rates were （78.5 ± 4.2 ）% and the A values measured by spectrophotometer were 0.46 ±0.05 on the 12 th day in the transfected group;the difference was significant as compared with untransfected MSCs （P 〈 0.05）. There was expression of the ADRP protein in the adipocytes,which was signifcantly higher in the transfected group than in the untransfected group. CONCLUSION： The overexpression of PPARγ can reinforce the ability of MSCs to differentiateinto adipocytes, which may be related to the increased expression of ADRP.
出处
《第四军医大学学报》
北大核心
2008年第20期1825-1828,共4页
Journal of the Fourth Military Medical University
基金
湖南省卫生厅科研基金(B2006-116)
