CAJ | 학술논문

目的观察抵抗素结合多肽(RBP)对3T3-L1脂肪细胞分化、脂代谢及葡萄糖转运体4(GLUT-4)基因表达的影响.方法构建大鼠抵抗素真核表达载体并转染3T3-L1前体脂肪细胞,获得稳定表达抵抗素基因细胞株;采用台盼蓝排斥试验,确定理想的RBP干预浓度,于诱导细胞分化第0天加入培养液;采用油红O染色,观察脂肪细胞分化及脂质积聚情况;采用RT-PCR技术检测脂肪细胞分化标志基因及GluT-4基因表达变化;采用全自动生化仪比色法,检测脂肪细胞内TG和游离脂肪酸FFAs含量的变化.结果 (1) RBP浓度10-12mol/L时,脂肪细胞活细胞数比例较高,且细胞形态无明显改变.(2)RBP对正常脂肪细胞分化进程无明显影响,RBP虽未影响抵抗素稳定表达脂肪细胞内脂滴的出现时间,但细胞内脂滴的数目明显减少.(3)RBP对正常脂肪细胞分化标志基因及抵抗素稳定表达细胞分化早期标志基因Pref-1的表达无明显影响,但明显下调抵抗素稳定表达细胞分化中晚期标志基因C/EBPα和FAS的表达水平.(4)RBP对正常脂肪细胞内TG、FFAs含量无影响,但可显著降低抵抗素稳定表达脂肪细胞内的TG、FFAs含量.(5)RBP干预对正常脂肪细胞及抵抗素稳定表达脂肪细胞中GluT-4基因的表达水平均无显著影响.结论 RBP对正常3T3-L1脂肪细胞的分化、脂代谢、GluT-4基因表达均无明显影响,但能有效拮抗抵抗素基因,显著促进3T3-L1脂肪细胞分化及脂代谢.
목적 관찰저항소결합다태(RBP)대3T3-L1지방세포분화、지대사급포도당전운체4(GLUT-4)기인표체적영향.방법 구건대서저항소진핵표체재체병전염3T3-L1전체지방세포,획득은정표체저항소기인세포주;채용태반람배척시험,학정이상적RBP간예농도,우유도세포분화제0천가입배양액;채용유홍O염색,관찰지방세포분화급지질적취정황;채용RT-PCR기술검측지방세포분화표지기인급GluT-4기인표체변화;채용전자동생화의비색법,검측지방세포내TG화유리지방산FFAs함량적변화.결과 (1) RBP농도10-12mol/L시,지방세포활세포수비례교고,차세포형태무명현개변.(2)RBP대정상지방세포분화진정무명현영향,RBP수미영향저항소은정표체지방세포내지적적출현시간,단세포내지적적수목명현감소.(3)RBP대정상지방세포분화표지기인급저항소은정표체세포분화조기표지기인Pref-1적표체무명현영향,단명현하조저항소은정표체세포분화중만기표지기인C/EBPα화FAS적표체수평.(4)RBP대정상지방세포내TG、FFAs함량무영향,단가현저강저저항소은정표체지방세포내적TG、FFAs함량.(5)RBP간예대정상지방세포급저항소은정표체지방세포중GluT-4기인적표체수평균무현저영향.결론 RBP대정상3T3-L1지방세포적분화、지대사、GluT-4기인표체균무명현영향,단능유효길항저항소기인,현저촉진3T3-L1지방세포분화급지대사.
Objective To investigate the effect of RBP on 3T3-L1 adipocyte differentiation,lipid metabolism and glucose transporter 4(GLUT-4)gene expression.Methods We constructed an expression vector for rat resistin gene and transfected it into 3T3-L1 adipocytes.RBP was added to the medium of 3T3-L1 adipocytes or resistin-overexpressing adipocytes on day 0 of differentiation.Cell differentiation and lipid accumulation were determined by oil red O staining.The mRNA expressions of differentiation marker genes(pref-1,C/EBPα,FAS)and GLUT-4 gene were evaluated by RT-PCR.Triglyceride(TG)and free fatty acids(FFAs)in adipocytes were measured by colorimetric kit.Results(1)When 10-12mol/L RBP was applied,the percent of living cells was high and the shape was unchanged.(2)RBP had no effect on the differentiation of normal adipocytes,but significantly decreased the number of lipid droplets in resistin-overexpressing adipocytes without affecting the lipid droplets-presenting day.(3)C/EBPα and FAS expressions in resistin-overexpressing adipocytes were down-regulated after RBP was applied,without changing their expressions in normal adipocytes.(4)RBP had no effect on the cellular TG and FFAs levels in normal cells,whereas it can significantly decrease the levels in resistin-overexpressing adipocytes.(5)There was no difference in the expression of GLUT-4 gene between 3T3-L1 adipocytes and RBP-applied cells.Conclusions(1)RBP has no effect on the cell differentiation and lipid metabolism in normal 3T3-L1 adipocytes.(2)RBP can inhibit the cell differentiation and lipid metabolism of resistin-overexpressing 3T3-L1 cells.(3)RBP has no effect on the expression of GLUT-4 gene.