IL-1β/JNK通路在七氟烷诱发大鼠离体海马神经元程序性坏死中的作用  

Role of IL-1β/JNK pathway in sevoflurane-induced necroptosis of rat hippocampal neurons in vitro

作　　者：张琦 李亚南[2] 梁诗瑶 尹春平[2] 赵娟[4] 王秋筠[2] Zhang Qi;Li Yanan;Liang Shiyao;Yin Chunping;Zhao Juan;Wang Qiujun(Department of Anesthesiology,Children's Hospital of Hebei Province,Shijiazhuang 050030,China;Department of Anesthesiology,Third Hospital of Hebei Medical University,Shijiazhuang 050051,China;College of Veterinary and Agricultural Sciences,University of Melbourne,Melbourne VIC3010,Australia;Experimental Centre for Teaching of Hebei Medical University,Shijiazhuang 050011,China)

机构地区：[1]河北省儿童医院麻醉科,石家庄050030 [2]河北医科大学第三医院麻醉科,石家庄050051 [3]墨尔本大学兽医和农业科学学院,墨尔本VIC3010,澳大利亚 [4]河北医科大学教学实验中心,石家庄050011

出　　处：《中华麻醉学杂志》2021年第12期1463-1466,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金面上项目(81771134);河北省自然科学基金面上项目(H201820-6305)。

摘　　要：目的评价IL-1β/c-Jun氨基末端激酶(JNK)通路在七氟烷诱发大鼠离体海马神经元程序性坏死中的作用。方法原代培养SD大鼠胎鼠的海马神经元,分别接种于96孔板(1×10^(4)个/ml,200μl/孔)和6孔板(1×10^(6)个/ml,2 ml/孔)中,培养7 d时,采用随机数字表法分为3组(n=20):对照组(C组)、七氟烷组(S组)和IL-1受体拮抗剂组(I组)。C组常规培养,I组加入IL-1受体拮抗剂IL-1ra 1μg/μl孵育30 min后,将S组和I组置于含2%七氟烷的培养箱中,37℃培养5 h。收集神经元,倒置显微镜下观察神经元形态,采用流式细胞术检测神经元程序性坏死率,MTT法检测神经元活力,采用Western blot法检测IL-1β、IL-1受体Ⅰ型(IL-1RI)、IL-1受体辅助蛋白(IL-1RAcP)、磷酸化JNK(p-JNK)、受体相互作用蛋白1(RIP1)、RIP3和磷酸化人混合系列蛋白激酶样结构域(p-MLKL)的表达水平。结果与C组比较,S组神经元活力降低,程序性坏死率升高,IL-1β、IL-1RI、IL-1RAcP、p-JNK、RIP1、RIP3和p-MLKL表达上调(P<0.05);与S组比较,I组神经元活力升高,程序性坏死率降低,IL-1β、IL-1RI、IL-1RAcP、p-JNK、RIP1、RIP3和p-MLKL表达下调(P<0.05)。C组神经元形态未见明显异常,S组神经元胞体皱缩,突起断裂及突起间网络稀疏;I组神经元胞体圆润,形态接近正常。结论七氟烷诱发大鼠离体海马神经元程序性坏死的机制与激活IL-1β/JNK通路有关。Objective To evaluate the role of interleukin 1β(IL-1β)/c-Jun N-terminal kinase(JNK)pathway in sevoflurane-induced necroptosis of rat hippocampal neurons in vitro.Methods Primarily cultured hippocampal neurons of Sprague-Dawley rat fetuses were inoculated into 96-well plates(cell density:1×10^(4)cells/ml,200μl/hole)and 6-well plates(cell density:1×10^(6)cells/ml,2 ml/hole).The cells were divided into 3 groups(n=20 each)using a random number table method after being cultured for 7 days:control group(group C),sevoflurane group(group S)and IL-1 receptor antagonist group(group I).Group C received routine culture,IL-1 receptor antagonist IL-1ra 1μg/μl was added,and the cells were incubated for 30 min in group I,and in addition the cells were placed in the incubator containing 2%sevoflurane and cultured for 5 h at 37℃in S and I groups.The cells were collected for microscopic examination of the morphology of neurons(with an inverted microscope)and for determination of the cell necroptosis rate(by flow cytometry),cell viability(by MTT method),and expression of IL-1β,interleukin-1 receptor type I(IL-1RI),interleukin-1 receptor accessory protein(IL-1RAcP),phosphorylated JNK(p-JNK),receptor-interacting protein 1(RIP1),RIP3 and phosphorylated mixed lineage kinase-like(p-MLKL)(by Western blot).Results Compared with group C,the cell viability was significantly decreased,the necroptosis rate was increased,and the expression of IL-1β,IL-1RI,IL-1RAcP,p-JNK,RIP1,RIP3 and p-MLKL was up-regulated in group S(P<0.05).Compared with group S,the cell viability was significantly increased,the necroptosis rate was decreased,and the expression of IL-1β,IL-1RI,IL-1RAcP,p-JNK,RIP1,RIP3 and p-MLKL was down-regulated in group I(P<0.05).There was no obvious abnormality in the morphology of neurons in group C.The cell body of neurons was shrunk,the processes were broken,and the network between processes was sparse in group S.The cell body was round,and the morphology was close to normal in group I.Conclusion The mechanism by which sev

关 键 词：麻醉药 吸入 海马 神经元 坏死 白细胞介素1Β JNK丝裂原活化蛋白激酶类 

分 类 号：R614[医药卫生—麻醉学]