CFTR激动剂forskolin对脑缺血再灌注诱导的神经元凋亡的影响  

Effects of CFTR agonist forskolin on neuronal apoptosis induced by cerebral ischemia reperfusion

作　　者：朱耀斌[1] 张雅娉[2] 李志强[1] 丁楠[1] 伊寒露 沈磊[1] 赵宇东[1] 叶赞凯 ZHU Yaobin;ZHANG Yaping;LI Zhiqiang;DING Nan;YI Hanlu;SHEN Lei;ZHAO Yudong;YE Zankai(Department of Pediatric Cardiac Surgery,Beijing Children's Hospital of Capital Medical University,National Center for Children's Health,Beijing100045,China;Department of Cardiac Surgery,Beijing Friendship Hospital of Capital Medical University,Beijing100037,China)

机构地区：[1]国家儿童医学中心首都医科大学附属北京儿童医院小儿心脏外科,北京100045 [2]首都医科大学附属北京友谊医院心脏外科,北京100037

出　　处：《中华实用诊断与治疗杂志》2020年第4期339-342,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：北京市卫生系统高层次卫生技术人才-学科骨干(2015-3-048);北京市卫生系统高层次卫生技术人才-学科骨干(2015-3-051);北京市卫生系统高层次卫生技术人才-学科骨干(2014-3-043);北京市自然科学基金(7184204);北京市自然科学基金(7182042)北京市医管局培育计划(PX2016046);北京市科委首都临床特色应用研究(Z171100001017048);北京市医院管理局儿科协同发展中心专项经费资助(XTYB201819)。

摘　　要：目的探讨囊性纤维化跨膜传导调节因子(cystic fibrosis transmembrane conductance regulator,CFTR)激动剂forskolin(FSK)对脑缺血再灌注(ischemia reperfusion,IR)诱导的神经元凋亡的影响。方法将48只小鼠随机分为对照组(假手术)、FSK组(假手术+腹腔注射FSK)、IR组(建立IR模型)、FSK+IR组(建立IR模型+腹腔注射FSK),每组12只。采用HE染色观察小鼠海马和皮质组织细胞形态学变化,采用TUNEL染色观察小鼠海马组织神经元凋亡情况,采用比色法测定小鼠神经元caspase-3、caspase-8、caspase-9活性。结果对照组和FSK组小鼠海马及皮质神经元染色均匀,胞质透明,细胞核大而圆,核仁清晰;IR组小鼠海马及皮质神经元结构破坏,细胞核浓染、固缩,呈神经元凋亡改变;FSK+IR组海马和皮质神经元细胞核浓染、固缩较IR组轻。生物素化-dUTP TUNEL染色中细胞核中出现棕黄色颗粒的神经元为凋亡神经元,荧光四甲基罗丹明-dUTP TUNEL染色细胞核中出现粉红色荧光的神经元为凋亡神经元;IR组和FSK+IR组小鼠海马神经元凋亡数量[(57.64±12.68)、(26.49±6.37)个/视野]高于对照组[(5.61±1.14)个/视野]和FSK组[(5.72±1.24)个/视野](P<0.05)。IR组和FSK+IR组小鼠海马神经元caspase-3、caspase-8、caspase-9活性(2.86±0.55、1.97±0.62、2.38±0.64,1.84±0.37、1.32±0.41、1.41±0.53)高于对照组(1.14±0.17、0.84±0.15、0.95±0.21)和FSK组(1.23±0.22、0.81±0.13、0.97±0.20)(P<0.05),FSK+IR组小鼠海马神经元凋亡数量和caspase-3、caspase-8、caspase-9活性低于IR组(P<0.05),对照组小鼠海马神经元凋亡数量及caspase-3、caspase-8、caspase-9活性与FSK组比较差异无统计学意义(P>0.05)。结论激活CFTR氯离子通道可降低IR小鼠脑组织神经元细胞凋亡,对IR小鼠脑组织神经元细胞具有保护作用。Objective To investigate the effect of forskolin(FSK),an agonist of cystic fibrosis transmembrane conductance regulator(CFTR),on neuronal apoptosis induced by cerebral ischemia reperfusion(IR).Methods Forty-eight mice were randomly divided into sham operation group,FSK group(sham operation+intraperitoneal injection of FSK),IR group(establishing IR model)and FSK+IR group(establishing IR model+intraperitoneal injection of FSK),with 12 mice in each group.HE staining was used to observe the morphological changes of cells in the hippocampus and cortex of mice.TUNEL staining was used to observe the neuronal apoptosis in the hippocampus.Colorimetry was used to determine the activities of caspase-3,-8 and-9 in the neurons.Results HE staining was uniform,the cytoplasm was transparent,the nucleus were large and round,and the nucleolus were clear in the hippocampus and cortex of sham operation group and FSK group;the neuronal structure was destroyed,the nucleus were densely stained and pyknotic,and the neuron was apoptotic in the hippocampus and cortex of IR group;the nucleus staining and pyknosis in the hippocampus and cortex of FSK+IR group were lighter than those of IR group.The apoptotic neurons were found in brown-yellow particles in biotinylated-dUTP TUNEL stain,and in pink fluorescent tetramethylrhodamine-dUTP TUNEL stain.The count of hippocampal apoptotic neurons was higher in IR group((57.64±12.68)/fields)and FSK+IR group((26.49±6.37)/fields)than that in sham operation group((5.61±1.14)/fields)and FSK group((5.72±1.24)/fields)(P<0.05).The activities of caspase-3,-8 and-9 were higher in IR group(2.86±0.55,1.97±0.62,2.38±0.64)and FSK+IR group(1.84±0.37,1.32±0.41,1.41±0.53)than those in sham operation group(1.14±0.17,0.84±0.15,0.95±0.21)and FSK group(1.23±0.22,0.81±0.13,0.97±0.20)(P<0.05).The count of hippocampal apoptotic neurons and the activities of caspase-3,-8 and-9 were lower in FSK+IR group than those in IR group(P<0.05),and showed no significant differences between sham operation group and FSK

关 键 词：缺血再灌注 囊性纤维化跨膜传导调节因子 FORSKOLIN 脑 神经元 凋亡 小鼠 

分 类 号：R965[医药卫生—药理学]