半胱氨酸天冬氨酸蛋白酶-1活化在胆红素神经毒性中的作用研究

发布时间：2018-03-20 13:47

本文选题：胆红素　切入点：神经毒性　出处：《第三军医大学学报》2017年18期 　论文类型：期刊论文

【摘要】：目的明确半胱氨酸天冬氨酸蛋白酶-1(caspase-1)活化是否参与胆红素神经毒性的发生机制;VX-765抑制caspase1活化是否能抑制胆红素神经毒性,发挥神经保护作用。方法建立胆红素脑病动物模型,Western blot检测脑组织caspase-1蛋白的表达,ELISA法检测炎症因子IL-1β的水平,免疫荧光染色检测脑组织中GFAP蛋白的表达;VX-765干预后动态观察各组新生鼠的神经系统临床表现,记录新生鼠体质量变化,评估生活能力。原代培养大鼠皮层星型胶质细胞分为胆红素组、VX-765组、对照组:改良MTT法检测细胞存活率,Western blot检测细胞caspase-1蛋白的表达,ELISA法检测培养液上清IL-1β的水平。结果胆红素脑病动物模型,建模后12 h,胆红素组较对照组,脑组织中活化型caspase-1表达增加(P0.05),IL-1β水平增高(P0.01),脑组织切片皮层区星型胶质细胞活化(P0.05)。与胆红素组相比,VX-765组新生鼠建模后异常神经系统表现减少(P0.01),生活能力改善(P0.05)。原代培养大鼠皮层星型胶质细胞,胆红素干预6h后,活化型caspase-1表达显著高于对照组(P0.05);与胆红素组相比,VX-765干预可抑制caspase-1活化(P0.05),提高细胞存活率(P0.05),减少培养液上清IL-1β的释放(P0.01)。结论胆红素可诱导活化型caspase-1表达增加;VX-765抑制caspase-1活化可减轻胆红素神经毒性,提高原代培养星型胶质细胞存活率,减少炎症因子释放。
[Abstract]:Objective to make clear the caspase -1 (caspase-1) activation mechanism is involved in bilirubin neurotoxicity; inhibit the activation of VX-765 inhibits caspase1 bilirubin neurotoxicity, play a neuroprotective effect. Methods to establish the animal model of bilirubin encephalopathy, caspase-1 protein expression of Western blot in brain tissue was detected, detection of inflammatory cytokine of IL-1 by ELISA, immunofluorescence staining to detect the expression of GFAP protein in brain tissue; clinical manifestations of nervous system and prognosis were dynamically observed in neonatal rat VX-765, records of body mass change in neonatal rats, evaluation of life ability. Primary cultured rat cortical type star glial cells into bilirubin group, VX-765 group, control group: the cell viability was measured by modified MTT method the expression of Caspase-1 protein was detected by Western, blot, ELISA level was detected in culture supernatant of IL-1 beta bilirubin encephalopathy animal results. The model, after modeling 12 h, bilirubin group than in the control group, brain activation increases the expression of Caspase-1 (P0.05), increased IL-1 levels (P0.01), cerebral cortex tissue sections of activated astrocytes (P0.05). Compared with the bilirubin group, VX-765 rats after modeling abnormal neurological manifestations reduced (P0.01), improve the ability of life (P0.05). Primary cultured rat cortical glial cell type star, bilirubin 6h, activated the expression of Caspase-1 was significantly higher than that of the control group (P0.05); compared with the bilirubin group, VX-765 intervention can inhibit the activation of Caspase-1 (P0.05), improve the survival rate of cells (P0.05), reduced the supernatant of IL-1 beta release (P0.01). Conclusion bilirubin can induce increased expression of type caspase-1; inhibit the activation of VX-765 can reduce the bilirubin neurotoxicity of Caspase-1, improve primary cultured astrocytes survival rate, reduce the release of inflammatory factors.

【作者单位】：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;
【基金】：国家自然科学基金青年科学基金(81200459) 重庆市基础科学与前沿技术研究一般项目(CSTC2013jcyj A0020)~~
【分类号】：R722.1

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