cAMP-PKA-CREB信号通路在缺氧预处理减轻异丙酚诱导发育期大鼠中枢神经毒性中的作用  

Role of cAMP-PKA-CREB signaling pathway in hypoxic preconditioning-induced reduction of propofol-induced central neurotoxicity in developing rats

作　　者：肖飞[1] 梁羽冰[2] 吕靖[1] 谢玉波[1] 利莉[1] Xiao Fei;Liang Yubing;Lyu Jing;Xie Yubo;Li Li(Department of Anesthesiology,First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China;Department of Anesthesiology,Affiliated Tumor Hospital of Guangxi Medical University,Nanning 530021,China)

机构地区：[1]广西医科大学第一附属医院麻醉科,南宁市530021 [2]广西医科大学附属肿瘤医院麻醉科,南宁市530021

出　　处：《中华麻醉学杂志》2018年第11期1288-1292,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金(81373498,81560500);广西自然科学基金(2017-GXNSFBA198108);广西医科大学青年基金(GXMUYSF201518)。

摘　　要：目的评价环磷腺苷-蛋白激酶A-环磷腺苷反应元件结合蛋白(cAMP-PKA-CREB)信号通路在缺氧预处理减轻异丙酚诱导发育期大鼠中枢神经毒性中的作用。方法SPF级雄性SD大鼠70只,7日龄,体重10~15g,采用随机数字表法将其分为7组(n=10):生理盐水组(N组)腹腔注射等容量生理盐水;异丙酚组(P组)腹腔注射异丙酚50mg/kg,待翻正反射恢复再后追加50mg/kg;缺氧预处理+异丙酚组(HP组)缺氧预处理(氧浓度8%维持10min,然后置于空气中10min,循环5次)结束后2h时腹腔注射异丙酚(方法同P组);缺氧预处理+异丙酚+PKA抑制剂H89组(HPH组)侧脑室注射H895μmol/5μl,30min后处理同HP组;异丙酚+PKA激动剂SP-CAMP组(PS组)侧脑室注射SP-CAMP20nmol/5μl,30min后腹腔注射异丙酚(方法同P组);生理盐水侧脑室注射组(NI组)和5%二甲基亚砜侧脑室注射组(DI组)分别侧脑室注射5μl生理盐水和5%二甲基亚砜。待翻正反射恢复后立即处死,取海马组织,行HE染色,采用免疫组化法测定海马PKAc和磷酸化CREB(p-CREB)阳性细胞数,采用Westernblot法测定海马活化caspase-3、Bcl-2、Bax、PKAc、CREB和p-CREB表达水平。结果与N组比较,P组海马活化caspase-3和Bax表达上调,Bcl-2、PKAc和p-CREB表达下调,PKAc和p-CREB阳性细胞百分比降低(P<0.05),海马细胞排列紊乱,细胞萎缩;与P组比较,HP组和PS组海马活化caspase-3表达下调,Bcl-2、PKAc和p-CREB表达上调,PKAc和p-CREB阳性细胞百分比升高,HP组Bax表达下调(P<0.05),海马细胞排列整齐、胞浆丰富、胞核可见;与HP组比较,HPH组海马活化caspase-3表达上调,Bcl-2、PKAc和p-CREB表达下调,PKAc和p-CREB阳性细胞百分比降低(P<0.05),细胞排列紊乱、细胞缩小、核固缩。结论缺氧预处理减轻异丙酚诱导发育期大鼠中枢神经毒性的机制可能与激活cAMP-PKA-CREB信号通路有关。Objective To evaluate the role of cyclic adenosine monophosphate-protein kinase A-cAMP response element-binding protein(cAMP-PKA-CREB)signaling pathway in hypoxic preconditioning-induced reduction of propofol-induced central neurotoxicity in the developing rats.Methods A total of 70 SPF male Sprague-Dawley rats,aged 7 days,weighing 10-15 g,were divided into 7 groups(n=10 each)using a random number table method:normal saline group(N group),propofol group(P group),hypoxic preconditioning plus propofol group(HP group),hypoxic preconditioning plus propofol plus PKA inhibitor H89 group(HPH group),propofol plus PKA agonist SP-CAMP group(PS group),normal saline injected via the lateral cerebral ventricle group(NI group),and 5% dimethyl sulfoxide(DMSO)injected via the lateral cerebral ventricle group(DI group).In P group,propofol 50 mg/kg was intraperitoneally injected,and an increment of propofol 50 mg/kg was given after recovery of righting reflex.The equal volume of normal saline was given instead in N group.In HP group,hypoxic preconditioning(rats were subjected to 5 cycles of 10-min hypoxia of 8% O2 and 10-min normoxia of 21% O2)was performed,and propofol was intraperitoneally injected at 2 h after the end of hypoxic preconditioning and the method was similar to those previously described in P group.In HPH group,H89 5 μmol/5 μl was injected via the lateral cerebral ventricle,and 30 min later the other treatment was similar to those previously described in HP group.In PS group,SP-CAMP 20 nmol/5 μl was injected via the lateral cerebral ventricle,and 30 min later propofol was injected using the method previously described in P group.In NI and DI groups,5 μl normal saline and 5% DMSO were injected via the lateral cerebral ventricle,respectively.Rats were immediately sacrificed after the righting reflex was recovered,brains were removed and hippocampi were isolated and cut into sections which were stained with haematoxylin and eosin for determination of PKAc and p-CREB positive cells(by immuno-histochemistry)and expr

关 键 词：缺氧 缺血预处理 环AMP 环AMP依赖性蛋白激酶类 CAMP反应元件结合蛋白质 二异丙酚 药物毒性 

分 类 号：R614[医药卫生—麻醉学]