异氟醚和七氟醚对新生大鼠皮质凋亡以及Akt和Bcl-xl/Bad表达的不同影响  

Different Effects of Isoflurane and Sevoflurane on Apoptosis of Cortical Neuron and Expression of Akt and Bcl-xl/Bad in Neonatal Rats

作　　者：李玉娟[1] 柳垂亮[2] 王飞[1] 陈伟强[1] 张静[1] 彭书崚[1] 

机构地区：[1]中山大学孙逸仙纪念医院麻醉科,广东广州510120 [2]广东省中医院麻醉科,广东广州510120

出　　处：《中山大学学报（医学科学版）》2011年第3期291-296,共6页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家青年自然科学基金(30700787/C03030301);广东省自然科学基金(8151040701000062)

摘　　要：【目的】研究相同麻醉深度的异氟醚和七氟醚对新生大鼠皮质神经细胞凋亡以及Akt和Bcl-xl/Bad蛋白表达的影响。【方法】5窝出生后7d(P7)的新生大鼠,每窝取11只(总共55只),每窝随机分为异氟醚组(I组)4只,七氟醚组(S组)4只和对照组(C组)3只。各组分别吸入1.1%异氟醚或1.8%七氟醚或空气4h,在麻醉结束时每窝每组各取一只幼鼠经心脏穿刺抽血检测血气和血糖的变化(n=5);在麻醉结束后2h每窝每组各取1只幼鼠灌注取脑,免疫组织化学法检测顶叶皮质区(PC)Caspase-3表达(n=5);另外,C组在麻醉处理0h,I组和S组分别在麻醉2h,4h每窝各取一只幼鼠取新鲜脑皮质,Westernblot检测磷酸化Akt,以及Akt,Bcl-xl和Bad表达的变化(n=5)。组间资料的比较采用单因素方差分析。【结果】I组Caspase-3阳性细胞数为(27.12±5.22)个/mm2,较对照组[(3.15±0.59)个/mm2]增加751.11%(P<0.001),比S组[(9.05±1.76)个/mm2]增加199.67%(P<0.01);S组Caspase-3表达与对照组比较差异无统计学意义。I组磷酸化Akt表达在麻醉2h较对照组降低66.31%(P<0.01),4h呈回升趋势;S组在2h和4h与对照组无统计学差异。I组Bad表达在麻醉2h较对照组增加162.2%(P<0.05),S组Bad表达在2h和4h与对照组差异均无统计学意义。I组和S组在2h和4hBcl-xl表达与对照组差异均无统计学意义。I组Bcl-xl/Bad比值在麻醉2h较对照组降低42.85%(P<0.01),S组在2h和4h与对照组无统计学差异。【结论】0.5MAC异氟醚比七氟醚诱导更多新生大鼠大脑皮质神经细胞凋亡,异氟醚抑制Akt磷酸化并降低Bad表达,下调Bcl-xl/Bad比值可能是其诱导神经细胞凋亡的机制之一;七氟醚不会影响Akt磷酸化和Bcl-xl/Bad比值。[Objective] To investigate the effects of isoflurane and sevoflurane at the same depth of anesthesia on apoptosis of neuron in the parietal cortex and the expression of Akt and Bcl-xl/Bad proteins of cortex in neonatal rats.[Methods] Five litters of neonatal rats at postnatal day 7,eleven rats in each litter,（altogether fifty-five rats） were involved in this study.Rats in each litter were assigned randomly into three groups：4 rats in isoflurane group（I group）,4 rats in sevoflurane group（S group） and 3 rats in control group（C group）.The rats in I group,S group,or C group were separately exposed to 1.1% isoflurane or 1.8% sevoflurane（equivalent to 0.5 MAC for neonatal rats） or air for 4 h.One neonatal rat in each group each litter was punctured though heart at the end of anesthesia and blood was get to measure its pH and blood glucose（n = 5）.One neonatal rat in each litter was perfused at 2 h after isoflurane or sevoflurane exposure and their brain were embedded by paraffin.Caspase-3 positive expression in the parietal cortex of brain were detected by immunohistochemistry staing（n = 5）.Besides,the fresh cortex were dissected at 0 h in C group and at 2 h,4 h in I group and S group,phospho-Akt,Akt,Bcl-xl,and Bad protein expression were detected by Western blot（n = 5）.One wayANOV were used for data analysis among groups.[Result] The numbers of caspase-3 positive cells in the parietal cortex in I group were（27.12±5.22） per mm2,which increased by 751.11% when compared with C group [（3.15±0.59）per mm2,P0.001] and increased by 199.67% when compared with S group [（9.05±1.76） per mm2,P〈0.01],but there were no significant difference between C group and S group.The expression of phospho-Akt protein in the cortex decreased by 66.31%（P〈0.01） at 2 h and recovered at 4 h in I group when compared with C group,while there were no significant difference at 2 h and 4 h between S group and C group.The expression of Bad protein in the cortex increased by 162.2%（P〈0.05） at 2

关 键 词：异氟醚 七氟醚 脑 原癌基因蛋白质c-Akt蛋白 Bcl相关死亡蛋白 

分 类 号：R729[医药卫生—儿科]