首都医科大学学报 ›› 2007, Vol. 28 ›› Issue (3): 304-306.

• 专题报道 • 上一篇    下一篇

瘦素对大鼠肝细胞瘦素受体蛋白、基因表达及酪氨酸磷酸化的影响

郭启煜1, 高妍2, 丛琳3, 邵宁生4   

  1. 1. 海军总医院内分泌科;2. 北京大学第一医院内分泌科;3. 北京大学第三医院运动医学研究所;4. 军事医学科学院基础医学研究所
  • 收稿日期:2007-04-10 修回日期:1900-01-01 出版日期:2007-06-24 发布日期:2007-06-24

Effects of Leptin on Gene and Protein Expression and Tyrosine Phosphorylation of Leptin Receptor in Rat hepatocyte

Guo Qiyu1, Gao Yan2, Cong Lin3, Shao Ningsheng4   

  1. 1. Department of Endocrinology, Navy General Hospital;2. Department of Endocrinology, First Affiliated Hospitalof Beijing University;3. Institute of Sports Medicine, Third Affiliated Hospital of Beijing University;4. Instituteof Basic Medical Sciences, Academy of Military Medical Sciences
  • Received:2007-04-10 Revised:1900-01-01 Online:2007-06-24 Published:2007-06-24

摘要:

目的 观察瘦素对大鼠肝细胞瘦素受体蛋白、基因表达及酪氨酸磷酸化的影响。方法 分离、培养新生Sprague-Dawley大鼠肝脏细胞,与瘦素共同孵育12 h、24 h及36 h,提取蛋白后用Western blot法检测瘦素受体的蛋白水平的变化,用RT-PCR法检测大鼠肝细胞瘦素受体的mRNA水平,用免疫共沉淀法检测瘦素受体的酪氨酸磷酸化程度。结果 大鼠肝细胞的瘦素受体的蛋白水平在与瘦素共同孵育12 h、24 h和36 h后与对照组相比差异无统计学意义(P>0.05)。大鼠肝细胞瘦素受体的mRNA表达在孵育12 h后与对照组相比差异无统计学意义(P>0.05),孵育24 h及36 h后,与对照组及孵育12 h时相比,mRNA水平明显升高,差异有统计学意义(P<0.05)。经瘦素作用12 h后,对大鼠肝脏细胞瘦素受体酪氨酸磷酸化无显著影响(P>0.05),但孵育24 h及36 h后,大鼠肝脏细胞瘦素受体的酪氨酸磷酸化程度与对照组及瘦素作用12 h后相比均显著升高,差异有统计学意义(P<0.05)。结论 瘦素可促进大鼠肝细胞瘦素受体的mRNA表达并增强其酪氨酸磷酸化程度。

关键词: 瘦素, 瘦素受体, 信号转导, 酪氨酸磷酸化

Abstract:

Objective To investigate effects of leptin on gene and protein expression and tyrosine phosphorylation of leptin receptor in rat hepatocytes.Methods hepatocytes were isolated from new-born Sprague-Dawley rats,after incubation with leptin(5 nmol/L) for 12,24 and 36 h,Western bolt was used to assess the protein level of leptin-receptor.The mRNA expression level of leptin-receptor was assayed by RT-PCR.The tyrosine phosphorylation of leptin-receptor weas assayed by immuno-precipitation.Results After incubation with leptin(5 nmol/L) for 12,24 and 48 h,there was no difference in the protein contents of leptin receptors between leptin(5 nmol/L) treated and control hepatocytes(P>0.05).The mRNA content of leptin receptor in rat hepatocytes was similar to that of the control after incubation with leptin(5 nmol/L) for 12 h(P>0.05),but significantly decreased after 24 or 36 h(24 h: leptin(5 nmol/L) 0.37±0.05 vs control 0.33±0.03,P<0.05;;36 h: leptin(5 nmol/L) 0.36±0.04 vs control 0.32±0.01,P<0.05).The tyrosine phosphorylation of leptin-receptor was remarkably reduced after incubation with leptin(5 nmol/L) for 24 or 36 h(24 h: leptin(5 nmol/L) 0.30±0.05 vs control 0.27±0.05,P<0.05;36 h: leptin(5 nmol/L) 0.32±0.05 vs control 0.26±0.07,P<0.05).Conclusion Leptin(5 nmol/L) may up-regulate leptin receptor gene expression and its tyrosine phosphorylation in rat hepatocytes and may contribute to promoting the signal transduction of leptin itself.

Key words: leptin, leptin receptor, signal transduction, tyrosine phosphorylation

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