Bornfeldt K E, Arnqvist H J, Norstedt G
Department of Pharmacology, Faculty of Health Sciences, Linköping University, Sweden.
J Endocrinol. 1990 Jun;125(3):381-6. doi: 10.1677/joe.0.1250381.
The aim of this investigation was to study the regulation of insulin-like growth factor-I (IGF-I) gene expression in cultured rat aortic smooth muscle cells. Near-confluent cells were deprived of serum for 24 h and then exposed to IGF-I, insulin, serum, basic fibroblast growth factor (basic FGF), platelet-derived growth factor (PDGF-BB; consisting of B-chain homodimer) or GH for 24 h. Levels of IGF-I mRNA were measured by solution hybridization. The level of IGF-I mRNA was markedly decreased by 10% (v/v) newborn calf serum (78 +/- 4 (S.E.M.) % decrease), 1 nmol basic FGF/l (53 +/- 8%), and 1 nmol PDGF-BB/l (40 +/- 3%) when measured after 24 h. The effect of PDGF-BB was significant after 6 h and became more marked after 24 h. GH (1 nmol/l or 0.1 mumol/l) or insulin (1 nmol/l) had no effect after 24 h, whereas IGF-I (1 nmol/l) and insulin (10 mumol/l) increased IGF-I mRNA 64 +/- 20% and 46 +/- 14% respectively. The increase caused by IGF-I was demonstrated after 3 h, and was most marked after 24 h. Using Northern blot analysis of cultured aortic smooth muscle cells, IGF-I transcripts of 7.4, 1.7 and 1.1-0.8 kilobases were observed. Exposure of the cells to 10% serum, 1 nmol basic FGF/l or 1 nmol PDGF-BB/l for 48 h increased the cell number by 104 +/- 7%, 64 +/- 3% and 61 +/- 22% respectively, while IGF-I, insulin and GH had little effect.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究的目的是探讨培养的大鼠主动脉平滑肌细胞中胰岛素样生长因子-I(IGF-I)基因表达的调控。将接近汇合的细胞血清饥饿24小时,然后分别用IGF-I、胰岛素、血清、碱性成纤维细胞生长因子(碱性FGF)、血小板衍生生长因子(PDGF-BB;由B链同二聚体组成)或生长激素(GH)处理24小时。通过溶液杂交法检测IGF-I mRNA水平。24小时后检测发现,10%(v/v)新生小牛血清(降低78±4(标准误)%)、1 nmol/L碱性FGF(降低53±8%)和1 nmol/L PDGF-BB(降低40±3%)可使IGF-I mRNA水平显著降低。PDGF-BB在6小时后作用显著,24小时后作用更明显。24小时后,GH(1 nmol/L或0.1 μmol/L)或胰岛素(1 nmol/L)无作用,而IGF-I(1 nmol/L)和胰岛素(10 μmol/L)可分别使IGF-I mRNA增加64±20%和46±14%。IGF-I引起的增加在3小时后即可显现,24小时后最为明显。通过对培养的主动脉平滑肌细胞进行Northern印迹分析,观察到7.4、1.7和1.1 - 0.8千碱基的IGF-I转录本。细胞分别暴露于10%血清、1 nmol/L碱性FGF或1 nmol/L PDGF-BB 48小时后,细胞数量分别增加104±7%、64±3%和61±22%,而IGF-I、胰岛素和GH作用甚微。(摘要截选至250词)