Caspase-1活化在胆红素诱导的大鼠海马神经元损伤中的作用  

Role of caspase-1 activation in bilirubin-induced injury in cultured primary rat hippocampal neurons

作　　者：韦倩 冯洁[1] 何春梅 华子瑜[1] WEI Qian;FENG Jie;HE Chunmei;HUA Ziyu(Department of Neonatology, Children＇s Hospital of Chongqing Medical University, Ministry of Education Key Laboratory of Child Development and Disorders, China International Science and Technology Cooperation Base of Child Development and Critical Disorders, Chongqing Key Laboratory of Translational Medical Research in Cognitive Development and Learning and Memory Disorders, Chongqing 400014, China)

机构地区：[1]重庆医科大学附属儿童医院新生儿科//儿童发育疾病研究教育部重点实验室//儿童发育重大疾病国家国际科技合作基地//认知发育与学习记忆障碍转化医学重庆市重点实验室,重庆400014

出　　处：《南方医科大学学报》2018年第5期567-571,共5页Journal of Southern Medical University

基　　金：国家自然科学基金(81200459);重庆市基础科学与前沿技术研究一般项目(CSTC2013jcyjA0020)~~

摘　　要：目的研究半胱氨酸天冬氨酸蛋白酶-1(caspase-1)活化及VX-765在胆红素诱导大鼠海马神经元损伤中的作用。方法原代培养大鼠海马神经元分为胆红素组、对照组、VX-765组;胆红素组给予胆红素(50μmol/L),对照组给予同体积药物溶剂二甲基亚砜(DMSO),VX-765组在胆红素干预前1 h给予VX-765(50μmol/L);Western blot检测NLRP3、活化caspase-1表达,改良MTT、乳酸脱氢酶(LDH)释放率及台盼蓝染色检测细胞相对存活率及死亡率,ELISA法检测原代培养上清IL-18水平。结果胆红素诱导原代培养大鼠海马神经元3、6 h,NLRP3、活化caspase-1蛋白表达明显高于对照组(P<0.05)。VX-765干预后,活化caspase-1表达与胆红素组相比明显降低(P<0.05)。分组干预细胞24 h,VX-765组的相对存活率为(84.020±2.311)%,明显高于胆红素组(P<0.05),低于对照组(P<0.05);VX-765组LDH释放率为(10.780±1.577)%,明显低于胆红素组(P<0.05),高于对照组(P<0.05);VX-765组台盼蓝染色阳性率为(5.580±1.234)%,明显低于胆红素组(P<0.05),高于对照组(P<0.05)。结论在原代培养的大鼠海马神经元中,caspase-1活化参与胆红素神经毒性的发生,VX-765抑制其活化发挥神经保护作用。Objective To investigate the role of caspase-1 activation in bilirubin-induced neuronal injury and the protective effect of VX-765 against bilirubin-induced neurotoxicity in cultured primary rat hippocampal neurons. Methods Cultured primary rat hippocampal neurons were exposed to DMSO（control group）, 50 μmol/L bilirubin, or 50 μmol/L bilirubin 1 h after50 μmol/L VX-765 treatment. The expressions of NLRP3 and caspase-1 in the neurons were detected by Western blotting, and the relative cell survival and death rates were assessed with a modified MTT assay, lactate dehydrogenase assay and Typan blue staining. Interleukin-18（IL-18） concentration in the culture supernatant was measured using enzyme-linked immunosorbent assay（ELISA）. Results In cultured primary rat hippocampal neurons, bilirubin exposure for 3 and 6 h caused significant increases in the expressions of NLRP3 and activated caspase-1 compared with those in the control group（P〈0.05）.Pretreatment of the cells with VX-765 obviously suppressed bilirubin-induced activation of caspase-1（P〈0.05）. The relative survival rate of the neurons was（84.02±2.31）% in VX-765 intervention group, significantly higher than that in bilirubin group（P〈0.05） but lower than that in the control group（P〈0.05）; LDH release rate in VX-765 intervention group was（10.78±1.58）%,significantly lower than that in bilirubin group（P〈0.05） but higher than that in the control group（P〈0.05）. The cell death rate in VX-765 intervention group was（5.58±1.23）%, significantly lower than that in bilirubin group（P〈0.05） but higher than that in the control group（P〈0.05）. Conclusion In cultured primary rat hippocampal neurons, caspase-1 activation plays a role in bilirubin-induced neurotoxicity, and VX-765 treatment provides protection against bilirubin-induced neuronal injury by inhibiting caspase-1 activation.

关 键 词：胆红素 神经毒性 海马神经元 CASPASE-1 VX-765 

分 类 号：R741[医药卫生—神经病学与精神病学]