本文選題：骨髓間充質(zhì)干細胞　切入點：成脂分化　出處：《重慶醫(yī)科大學學報》2016年10期

【摘要】：目的:探討炎癥因子轉(zhuǎn)化生長因子(transforming growth factor-α,TNF-α)對SD大鼠骨髓間充質(zhì)干細胞(bone marrow mesenchymal stem cells,BMSCs)成脂分化的影響及相關(guān)的分子機制。方法:將生長良好的第3代(P3)SD大鼠BMSCs分為對照組(Con組)、成脂誘導組(Ad組)和10 ng/m L TNF-α干預成脂誘導組(Ad+TNF-α組),每組各接種5塊6孔板,培養(yǎng)14 d后,收集各組細胞的RNA和蛋白質(zhì)。于第14天時間點處進行油紅O染色和異丙醇萃取,酶標儀490 nm處檢測吸光度(absorbance,A)值,觀察各組成脂分化的程度。q RT-PCR檢測各組基因Wnt10b、前體脂肪細胞因子-1(preadipocyte factor-1,Pref-1)、CCAAT區(qū)增強子結(jié)合蛋白-α(CCAAT/enhancer-binding protein-α,C/EBP-α)、過氧化物酶體增殖物激活受體-γ(peroxisome proliferator-activated receptor-γ,PPAR-γ)、脂肪酸結(jié)合蛋白-4(fatty acid binding protein-4,FABP-4)的m RNA表達水平。Western blot檢測第14天3組基因β聯(lián)蛋白(β-catenin)、PPAR-γ、C/EBP-α、FABP-4的蛋白表達水平。結(jié)果:TNF-α可以明顯抑制SD大鼠BMSCs的成脂分化。在Ad+TNF-α組,油紅O染色明顯低于Ad組,異丙醇萃取后A值檢測也明顯低于Ad組(0.360±0.035 vs.0.770±0.025,P=0.000)。q RT-PCR結(jié)果顯示,在Ad+TNF-α組,Wnt10b、Pref-1的m RNA水平(17.050±2.706,4.135±0.280)明顯高于Con組(P=0.019,P=0.003)和Ad組(0.575±0.065,0.514±0.060)(P=0.020,P=0.006),而C/EBP-α、PPAR-γ、FABP-4的m RNA水平(0.200±0.012,0.768±0.030,0.883±0.048)明顯低于Ad組(2.965±0.455,2.330±0.211,3.847±0.171)(P=0.019,P=0.000,P=0.001)。Western blot結(jié)果顯示,誘導14 d后,Ad+TNF-α組的C/EBP-α、PPAR-γ、FABP-4蛋白表達水平(0.586±0.013,0.356±0.008,0.118±0.002)明顯低于Ad組(1.082±0.018,0.840±0.112,1.094±0.038)(均P=0.000)。在Ad+TNF-α組,磷酸化β-catenin(P-β-catenin)的蛋白水平(0.648±0.005)明顯低于Ad組(3.376±0.211)(P=0.000)。結(jié)論:TNF-α抑制SD大鼠BMSCs成脂過程分化相的作用可能與Wnt/β-catenin信號通路活性的增加和Pref-1的高表達有關(guān),由此BMSCs的成脂分化停留在了前體脂肪細胞階段。
[Abstract]:Aim: to investigate the effect of transforming growth factor- 偽 (TNF- 偽) on lipid differentiation of bone marrow mesenchymal stem cells in SD rats and its molecular mechanism.Methods: the third generation of P3SD rats with good growth were divided into two groups: control group (Con group), adipogenic induction group (Ad group) and 10 ng/m L TNF- 偽 intervention group (Ad TNF- 偽 group). Each group was inoculated with 5 6-well plates and cultured for 14 days.The RNA and protein of each group were collected.Oil red O staining and isopropanol extraction were performed at the 14th day, and absorbency absorbency (A) was detected at 490 nm.瑙傚療鍚勭粍鎴愯剛鍒嗗寲鐨勭▼搴，

本文編號：1703672