胆碱能受体和肾上腺素能受体磷酸化的调节作用机制研究

发布时间：2018-06-27 16:44

本文选题：胆碱能受体 + 肾上腺素能受体　；参考：《吉林大学》2008年硕士论文

【摘要】： 本实验以纯化的大鼠脑毒蕈碱乙酰胆碱m2受体(mAChR2)和β2肾上腺素能受体膜蛋白(β2-AR)为研究对象,观察氨甲酰胆碱、阿托品和肝素分别对m2受体磷酸化过程的影响以及特布他林和肝素对β2-AR膜蛋白磷酸化过程的影响,探讨激活剂或抑制剂影响的G蛋白偶联受体激酶2 (GRK2)介导的胆碱能m2受体和β2-AR磷酸化的调节机制。探讨兴奋剂是如何使m2受体和β2-AR受体活化,引发涉及受体磷酸化的级联反应,进而影响蛋白激酶催化受体的磷酸化。分析这些受体的调节过程中涉及一些特异过程如受体的脱敏,内化和下调的起始机制。通过体外实验结果显示氨甲酰胆碱明显增强m2受体的磷酸化,而阿托品或肝素(GRK2抑制剂)完全阻断了m2受体的磷酸化。m2受体的磷酸化是依赖激活剂如氨甲酰胆碱作用发生的,呈明显的剂量依赖关系;由GRK2介导的β2-AR的激活剂特布他林以及肝素对β2-AR的磷酸化的检测结果显示,特布他林增强肾上腺素能受体的磷酸化,而肝素可完全阻断β2-AR的磷酸化;氨甲酰胆碱不能影响肾上腺素能受体的磷酸化,特布他林对mAChR2磷酸化也无明显影响。这些结果证实氨甲酰胆碱对mAChR2,特布他林对β2-AR磷酸化的增强作用是选择性的作用结果。进一步采用HEL299细胞磷酸化实验,结果显示氨甲酰胆碱可以增强HEL299细胞m2受体的磷酸化,阿托品可以抑制HEL299细胞m2受体的磷酸化但不如体外实验作用明显,揭示体内m2受体的磷酸化过程可能受激动剂和抑制剂共同调节。体外和体内实验证实了氨甲酰胆碱、特布他林和阿托品等一些m2受体和β2-AR的激活剂及抑制剂对M2受体和β2-AR磷酸化的调节作用。提示激活剂增强m2受体和β2-AR的磷酸化可能是这些受体活化一个重要部分,由此开始加速受体敏感性下调,受体耐受性增强。
[Abstract]:In this experiment, the effects of carbachol, atropine and heparin on the phosphorylation of M2 receptor and the effect of terbutaline and heparin on the phosphorylation of beta 2-AR membrane protein were investigated with the purified acetylcholine acetylcholine M2 receptor (mAChR2) and beta 2 adrenergic receptor membrane protein (beta 2-AR). The regulation mechanism of G protein coupled receptor kinase 2 (GRK2) mediated cholinergic M2 receptor and beta 2-AR phosphorylation. It is discussed how stimulant activates the M2 receptor and beta 2-AR receptor, triggering cascade reaction involving receptor phosphorylation and further affecting the phosphorylation of protein kinase catalytic receptor. Some specific processes, such as the desensitization, internalization and downregulation of receptors, show that methylcholine significantly enhances the phosphorylation of M2 receptors, while atropine or heparin (GRK2 inhibitor) completely blocks the phosphorylation of the phosphorylated.M2 receptor of the M2 receptor, which is dependent on the action of activator such as carbachol. The dose dependence; the results of GRK2 mediated beta 2-AR activator terbutaline and heparin phosphorylation of beta 2-AR showed that terbutaline enhanced the phosphorylation of adrenergic receptors, while heparin could completely block the phosphorylation of beta 2-AR; carbachol did not affect the phosphorylation of adrenergic receptors, terbutaline against mAChR 2 phosphorylation has no obvious effect. These results confirm that carbacholine is selective to the enhanced effect of mAChR2 and terbutaline on the phosphorylation of beta 2-AR. Further use of HEL299 cell phosphorylation experiments shows that carbachol can enhance the phosphorylation of M2 receptor in HEL299 cells, and atropine can inhibit M2 receptor in HEL299 cells. The phosphorylation of the body is not as effective as that in vitro. It reveals that the phosphorylation of M2 receptors may be regulated by agonists and inhibitors.
In vitro and in vivo experiments have confirmed the regulatory effects of some M2 receptors and beta 2-AR activators and inhibitors on the phosphorylation of M2 receptors and beta 2-AR, which suggest that activators enhance the phosphorylation of M2 receptors and beta 2-AR, which may be a critical part of these receptors, which begin to accelerate the sensitivity of the receptor. Intolerance, enhanced receptor tolerance.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R341

【相似文献】