中间蛋白在地塞米松体外诱导大鼠胚胎垂体生长激素细胞分化过程中作用的实验研究
程玉1，吕文明1，项莹2，胡恩喜1，张广会1，王伟军1
基金项目：教育部高等学校博士学科点专项科研基金（编号：20112307120031） 作者简介：程玉（1971-），男，副教授，主要研究方向：神经外科学.

（1. 哈尔滨医科大学附属第一医院神经外科，哈尔滨，150001； 2. 哈尔滨医科大学附属第二医院内分泌代谢科，哈尔滨，150086）
摘要：目的: 探究Ras、ERK、Akt中间蛋白在地塞米松体外诱导大鼠胚胎垂体生长激素细胞分化过程中的作用。方法：本课题利用大鼠胚胎垂体细胞的无血清原代细胞培养技术，在地塞米松诱导生长激素细胞分化的过程中，加入中间蛋白Ras、ERK和Akt的抑制剂Manumycin、U0126、LY294002进行预处理。利用免疫荧光、western-blot、放射免疫分析和MTT等技术对Ras、ERK以及Akt蛋白在糖皮质激素体外诱导生长激素细胞分化中的作用进行研究。结果：地塞米松能够显著提高生长激素阳性细胞百分比和生长激素的含量(P<0.01); Manumycin、U0126、LY294002抑制剂单独或联合预处理后，地塞米松诱导的生长激素阳性细胞百分比显著降低(P<0.01)，生长激素的含量亦出现降低（P<0.05）。结论：Ras、ERK和Akt蛋白是介导地塞米松诱导生长激素细胞分化的关键蛋白，在地塞米松诱导生长激素细胞分化过程中Ras、ERK和Akt蛋白发挥重要作用。
关键词：基础医学；地塞米松；生长激素细胞；分化；Ras/ERK；Ras/Akt
中图分类号：R329.2+1 20
Intermediate protein on Dexamethasone inducing Growth Hormone Cells Differentiation of Fetal Rat in vitro
Cheng Yu1, Lv Wenming1, Xiang Ying2, Hu Enxi1, Zhang Guanghui1, Wang Weijun1
(1. Department of Neurosurgery of First Affiliated Hospital,Harbin Medical 25 University,Harbin,150001; 2. Department of Endocrinnology of Second Affiliated Hospital,Harbin Medical University,Harbin,150086)
Abstract: Objective To explore the function of Ras、ERK and Akt in dexamethasone induces the differentiation of fetal rat pituitary somatotroph cells in vitro.Methods: This topic uses embryo   pituitary cells in vitro serum-free primary cell culture technique. The pituitary was harvested rendered into a fetal pituitary cell suspension. In the experiment Ras ERK and the Akt inhibitors, Manumycin U0126 and/or LY294002,were added to the suspension for pretreatment and cell culture was carried out using a medium containing Dex. After those, immunofluorescence, Western-blot, radioimmunoassay and MTT Technology is used in this experiment.Results: Dex can increase the percentage of GH-positive cells and GH content (P<0.01).After pretreatment with Manumycin U0126 and LY294002, the percentage of Dex-induced GH-positive cells decreased significantly (P<0.01), and the GH content was also reduced (P<0.05). Conclusion: Ras ERK and Akt are the key proteins during the process Dex induce GH cell differentiation. Ras, ERK and Akt proteins play an important role in the dexamethasone-induced growth hormone cell differentiation.
Key words: Basic medical；dexamthasone; GH cell; differentiation;Ras/ERK;Ras/Akt