前列腺素E_2调控MN9D细胞孤儿核受体Nurr1表达的研究  

Regulation of prostaglandin E_2 on orphan nuclear receptor Nurr1 expression in MN9D cells

作　　者：赵咏梅[1] 张海燕[2] 许秋岩[1] 吕风月[1] 

机构地区：[1]首都医科大学宣武医院神经变性病教育部重点实验室,北京100053 [2]首都医科大学细胞生物学系,北京100069

出　　处：《中国药理学通报》2009年第1期26-29,共4页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No90709011);北京市留学人员科技活动择优资助项目(2007年度)

摘　　要：目的观察前列腺素E2（prostaglandinE2，PGE2）对多巴胺（dopamine，DA）合成细胞系MN9D细胞内源性孤儿核受体Nurrl表达的作用，并研究Nurrl表达增高对酪氨酸羟化酶（tyrosinehydroxylase，TH）表达的影响，探讨Nurrl调控DA能神经元发育的机制。方法用100μg·L-1的PGE2作用于MN9D细胞2h至6h，观察细胞形态变化，并用免疫细胞化学染色及Westernblot方法检测PGE2作用前后MN9D细胞内源性Nurrl及TH表达的变化。结果①100μg·L-1的PGE2作用2、4及6h，MN9D细胞形态没有明显变化。②加入PGE：2、4及6h，MN9D细胞Nurrl抗体免疫荧光染色强度比未加PGE2作用的正常对照组细胞明显增强。MN9D细胞自身表达TH呈不均一性，PGE：作用2～6h，MN9D细胞中TH阳性染色细胞百分比与正常对照组细胞接近。③Westernblot结果显示，PGE：作用6h，MN9D细胞Nurrl蛋白表达比未加PGE，的正常对照组细胞明显增加（P〈0．05），而此时TH蛋白表达与正常对照组细胞接近。结论PGE，可在短时间内迅速明显激活MN9D细胞内源性Nurrl表达，但对TH表达没有影响。TH表达的激活或许还需要除Nurrl以外其它环境或因子的共同参与。Aim To investigate whether prostaglandin E2 （ PGE2 ） can increase endogenous Nurrl expression in a dopamine-synthesizing cell line （MN9D） with immature characteristics and to study whether tyrosine hydroxylase （TH） expression is up-regulated in re- sponse to Nurrl-overexpression, in order to investigate the role of Nurrl during dopaninergie neurons development. Methods MNgD cells were treated with 100 μg·L-1 PGE2 for 2 h to 6 h. The morphology changes of MN9D cells were observed under phase-contrast mi- croscope. The expression of Nurrl and TH in MN9D cells was analyzed by using immunohistochemical staining and Western blot analysis. Results （1） The morphology of MN9D cells did not change significantly following PGE2 treatment for 2 h, 4 h and 6h compared with that of MN9D cells left untreated. （2） Nurrl-positive staining in MN9D cells treated with PGE2 for 2 h,4 h and 6 h was much stronger than that of untreated cells while the percent of TH-positive MN9D cells after PGE2 treatment for 2 h, 4 h and 6 h was similar to that of untreated. （3） The expression of Nurrl protein in MN9D cells treatment With PGE2 for 6 h was signifi- cantly higher than that of untreated （ P 〈 0.05 ）, while the expression of TH protein in MN9D cells did not change significantly following Nurrl up-regulation induced by PGE2. Conclusion PGE2 up-regulates Nurrl protein expression in MN9D cells within short time （2-6 h） but TH expression doesn＇t elevate significantly. Up-regulation of Nurrl expression alone may be insufficient for activating TH expression in MN9D cells, but rather requires specific cellular （or neuronal） environments or cofactors.

关 键 词：前列腺素E2 NURR1 多巴胺能神经元 酪氨酸羟化酶 

分 类 号：R392.2[医药卫生—免疫学] R345.47[医药卫生—基础医学]