CAJ | 학술논문

目的探讨G蛋白偶联受体48(GRP48)在3T3-L1前体脂肪细胞诱导分化过程中的作用.方法体外培养3T3-L1前体脂肪细胞诱导分化为成熟脂肪细胞,在分化不同时段(第0～14天),采用Real-time PCR技术检测脂肪细胞中GPR48、过氧化物酶体增殖体激活受体γ2(PPAR2)和CCAAT增强子结合蛋白α(C/EBPα)基因信使核糖核酸(mRNA)的表达水平.利用SPSS 11.0统计软件对实验数据进行统计学分析.结果 GPR48基因在3T3-L1前体脂肪细胞诱导分化第2天和第3天表达显著上调,与诱导前期相比,与诱导前期相比差异均有统计学意义(t=4.12,P=0.015;t=6.21,P=0.003),分化第6～14天与分化前表达无差异.PPARγ2表达在诱导分化后明显上调,分化第6天达高峰,第10～ 14天持续处于较高水平并趋于稳定,与诱导前期相比各时段间表达水平差异均有统计学意义(t值为4.17 ～ 22.65,P均＜0.01).C/EBPα表达在诱导分化后明显上调,分化后第3天达高峰,第6～10天持续保持在较高水平,与诱导前期相比各时段表达水平差异均有统计学意义(t值为4.38 ～ 13.87,P均＜0.01),第14天趋于下调,与分化前比较无差异.GPR48基因表达高峰早于PPARγ2和C/EBPα.结论在3T3-L1脂肪细胞分化过程中PPARγ2和C/EBPα表达变化与脂肪细胞分化、脂质积聚过程相一致.GPR48基因表达高峰早于PPARγ2和C/EBPα,可能参与了脂肪细胞分化的早期过程.
목적 탐토G단백우련수체48(GRP48)재3T3-L1전체지방세포유도분화과정중적작용.방법 체외배양3T3-L1전체지방세포유도분화위성숙지방세포,재분화불동시단(제0～14천),채용Real-time PCR기술검측지방세포중GPR48、과양화물매체증식체격활수체γ2(PPAR2)화CCAAT증강자결합단백α(C/EBPα)기인신사핵당핵산(mRNA)적표체수평.이용SPSS 11.0통계연건대실험수거진행통계학분석.결과 GPR48기인재3T3-L1전체지방세포유도분화제2천화제3천표체현저상조,여유도전기상비,여유도전기상비차이균유통계학의의(t=4.12,P=0.015;t=6.21,P=0.003),분화제6～14천여분화전표체무차이.PPARγ2표체재유도분화후명현상조,분화제6천체고봉,제10～ 14천지속처우교고수평병추우은정,여유도전기상비각시단간표체수평차이균유통계학의의(t치위4.17 ～ 22.65,P균＜0.01).C/EBPα표체재유도분화후명현상조,분화후제3천체고봉,제6～10천지속보지재교고수평,여유도전기상비각시단표체수평차이균유통계학의의(t치위4.38 ～ 13.87,P균＜0.01),제14천추우하조,여분화전비교무차이.GPR48기인표체고봉조우PPARγ2화C/EBPα.결론 재3T3-L1지방세포분화과정중PPARγ2화C/EBPα표체변화여지방세포분화、지질적취과정상일치.GPR48기인표체고봉조우PPARγ2화C/EBPα,가능삼여료지방세포분화적조기과정.
Objective In order to investigate the role of GPR48 in 3T3-L1 preadipocytes differentiation,we assayed expression of G protein coupled receptor 48(GPR48),peroxisome proliferator-activated receptorγ2 (PPARγ2) and CCAAT Enhancer binding protein CEBP α(C/EBPα) genea during differentiation of 3T3-L1 preadipocytes.Methods 3T3-L1 cells were cultured,and induced to differentiate using 0.5 mmol/L 3-isobuty1-1-methyxanthine (IBMX),10tg/ml insulin,and 1 μmol/L dexamethasone.Real-time PCR was used to evaluate gene expression during 10 days' differentiation period.SPSS11.0 software was used for statistic analysis.Results The expression level of GPR48 reached peak at 2 days after induction of differentiation,significantly higher than that before induction (t =4.12,P =0.015;t =6.21,P =0.003).But the expression declined then and retuned to base level atter day 6.The expression level of PPARγ2 reached peak at day 3 after induction of differentiation,significantly higher than that before induction.The expression remained high thereafter(t =4.17 ～ 22.65,P ＜ 0.01).The expression level of C/EBPα reached peak at day 3 after induction,significantly higher than that before induction (t =4.12,P =0.015; t =6.21,P =0.003).The expression remained high thereafter(t =4.38 ～ 13.87,P ＜ 0.01).The expression peak of GPR48 was earlier than those of PPARγ2 and C/EBPα.Conclusion Expressions of GPR48,PPARγ2,and C/EBPαmRNA were upregulated after induction of differentiation.Both PPARγ2 and C/EBPα are involved in 3T3-L1 preadipocytes differentiation.GPR48 may play a role at the earlier time points after induction of differentiation.