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EFFECT OF INTERLEUKIN-1β ON GROWTH HORMONE GENE EXPREssION AND ITs POssIBLE MOLECULAR MECHANIsM IN RAT mtt/s sOMATOTROPH CELLs

引用

Chinese Medical sciences Journal 2008年第4期23卷 193-201页

作者： Feng-ying Gong Jie-ying Deng Yi-fan shi Department of Endocrinology Key Laboratory of Endocrinology of Ministry of Health Peking Union Medical College Hospital Chinese Academy of Medical Sciences ＆ Peking Union Medical College Beijing 100730

Objective To elucidate the effect of interleukin-1β (IL-1β) on human growth hormone (hGH) gene expression in a rat somatotropic pituitary cell line mtt/s. Methods stably transfected mtt/s cells were firstly established by transfecting 484-Luc1 plasmid which contained hGH gene promoter -484 to +30 bp and luciferase reporter gene. The effect of IL-1β on hGH gene expression was determined by assaying the luciferase activities. RT-PCR method was also used to determine whether IL-1 recepor mRNA was expressed in mtt/s cells. Results The 103 U/mL IL-1β stimulated secretion and synthesis of GH, and promoted the 5’-promoter activity of GH gene in stably transfected mtt/sGL cells with the action of 1.38 times above the control. Among inhibitors of signaling transduction pathways, mitogen-activated protein kinase kinase (MAPKK/MEK) inhibitor PD98059 (40 μmol/L) and p38 mitogen-activated protein kinase (MAPK) inhibitor sB203580 (5 μmol/L) completely blocked the stimulatory effect of IL-1β, and phosphatidylinositol-3-kinase (PI3-K) inhibitor LY294002 partly abolished the effect of IL-1β. Western blot analysis further confirmed the activation of phosphorylated MEK and p38 MAPK in mtt/sGL cells. Neither over-expression of Pit-1 nor inhibition of Pit-1 expression affected induction of hGH promoter activity by IL-1β. A series of deletion constructs of hGH promoter were created to identify the DNA sequence that mediated the effect of IL-1β, and results showed that the stimulatory effect of IL-1β was abolished following deletion of the -196 to -132 bp fragment. Conclusions IL-1β promotes GH secretion and synthesis in rat mtt/s somatotroph cells. The stimulatory effect of IL-1β on hGH gene promoter appears to require the activation of MEK, p38 MAPK, PI3-K, and a fragment of promoter sequence that spans the –196 to –132 bp of the gene, but it may be unlinked with Pit-1 protein.

关键词：白介素生长激素因子促进剂 mtt/s 细胞蛋白质

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