大鼠海马β-肾上腺素受体和烟碱受体亚型在烟碱激活p44/42MAPK中的作用  

Roles of β-adrenergic receptor and subtypes of nicotinic receptor in phosphorylation of p44/42 MAPK induced by nicotine in rat hippocampus

作　　者：王勇[1] 朱小南[2] 余剑平[2] 黄晓卉[2] 陈汝筑[2] 

机构地区：[1]第一军医大学珠江医院药剂科,广州510282 [2]中山大学基础医学院药理学教研室,广东广州510080

出　　处：《中国药理学与毒理学杂志》2004年第2期93-97,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：国家卫生部科研基金资助项目 (98 1 0 86 )~~

摘　　要：目的　为探讨烟碱对海马脑片p4 4/ 4 2MAPK磷酸化的影响 ,以及 β 肾上腺素受体 (β AR)、烟碱受体亚型在其中的作用。方法　将 4 0 0 μm厚的海马脑片置于预先经 95 %O2 和 5 %CO2 饱和的 2 8℃人工脑脊液中预孵 90min后分别加入普萘洛尔、α 银环蛇毒素和美卡拉明 ,30min后再加入烟碱 ,使其终浓度分别为 1和 10 0 μmol·L- 1。于烟碱作用 90min时 ,运用免疫印迹法检测这 3个受体阻断剂对烟碱引起的p4 4/ 4 2MAPK磷酸化的抑制效果。结果　烟碱对p4 4/ 4 2MAPK磷酸化的作用与烟碱浓度有关 ,10 0 μmol·L- 1烟碱比 1μmol·L- 1烟碱作用强。美卡拉明单独作用可阻断 1μmol·L- 1烟碱和部分阻断10 0 μmol·L- 1烟碱引起的p4 2MAPK磷酸化 ;α 银环蛇毒素只能阻断 10 0 μmol·L- 1烟碱引起的p4 2 /4 4MAPK磷酸化 ;但同时使用这两种烟碱受体阻断剂时 ,两种浓度烟碱引起的p4 4/ 4 2MAPK磷酸化均可被完全阻断 ;普萘洛尔对 1μmol·L- 1烟碱引起的p4 4MAPK磷酸化和 10 0 μmol·L- 1烟碱引起的p4 2MAPK磷酸化仅有部分阻止作用。结论　在烟碱激活的p4 4/ 4 2MAPK中 ,不同烟碱受体亚型在不同烟碱浓度中起作用 ;β AR仅参与烟碱引起的p4 4/4 2MAPK部分磷酸化。AIM To investigate the roles of β adrenergic receptor(β AR) and subtypes of nicotinic receptor in phosphorylation of p44/42MAPK induced by nicotine. METHODS Hippocampal slices(400 μm thick) obtained from Sprague Dawley rat brain were preincubated in artificial cerebrospinal fluid(ACSF) previously saturated with 95% O 2 and 5% CO 2 at 28℃ for 90 min, and then 10 μmol·L -1 of α bungarotoxin(α BGT), mecamylamine and propranolol were added respectively, 30 min after the addition, 1 or 100 μmol·L -1 nicotine was added into ACSF respectively. After incubation with nicotine for 90 min, phosphorylation of p44/42MAPK of the samples were determined by Western blotting. RESULTS Diversity of phosphorylation of p44/42MAPK arose from the concentration of nicotine, the increase in phosphorylation of p42MAPK induced by 100 μmol·L -1 nicotine was more significant than that by 1 μmol·L -1 nicotine. Mecamylamine could completely block the activation of p42MAPK induced by 1 μmol·L -1 nicotine and partially block that by 100 μmol·L -1 nicotine. In contrast, α BGT only blocked the activation of p44/42MAPK induced by 100 μmol·L -1 nicotine. However, α BGT companied with mecamylamine could eliminate the activation of p44/42MAPK by both of 1 and 100 μmol·L -1 nicotine. The phosphorylation of p44MAPK by 1 μmol·L -1 nicotine and that of p42MAPK by 100 μmol·L -1 nicotine could be partially blocked by propranolol. CONCLUSION The subtypes of nicotinic receptor contribute to the effects of different concentration of nicotine on the phosphorylation of p44/42MAPK. Partial phosphorylation of p44/42MAPK induced by nicotine involves the activation of β AR.

关 键 词：海马 受体 烟碱 受体 肾上腺素 烟碱 有丝分裂素激活蛋白激酶类 

分 类 号：R96[医药卫生—药理学]