论文部分内容阅读
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(P<0.05),IL-1β水平增高(P<0.01),脑组织切片皮层区星型胶质细胞活化(P<0.05)。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(P<0.01),生活能力改善(P<0.05)。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(P<0.05);与胆红素组相比,VX-765干预可抑制caspase-1活化(P<0.05),提高细胞存活率(P<0.05),减少培养液上清IL-1β的释放(P<0.01)。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。
Objective To determine whether activation of caspase-1 is involved in the neurotoxicity of bilirubin. VX-765 inhibits the activation of caspase 1 and inhibits the neurotoxicity of bilirubin and plays a neuroprotective role. Methods The animal model of bilirubin encephalopathy was established. The expression of caspase-1 protein in brain tissue was detected by Western blot. The level of IL-1β was detected by ELISA. The expression of GFAP protein in brain tissue was detected by immunofluorescence staining. Observe the clinical manifestations of the nervous system in neonatal rats in each group, record the changes of newborn rats’ body weight, and evaluate the living ability. Primary cultured rat cortical astrocytes were divided into bilirubin group, VX-765 group, control group: cell viability was detected by modified MTT method, the expression of caspase-1 protein was detected by Western blot, Supernatant IL-1β levels. Results In the bilirubin encephalopathy animal model, the expression of activated caspase-1 in the bilirubin group was increased (P <0.05), the level of IL-1β was increased (P <0.01) Activation of astrocytes in the sectioned cortex (P <0.05). Compared with the bilirubin group, the expression of abnormal nervous system decreased (P <0.01) and the viability improved in the VX-765 group (P <0.05). Compared with control group, the expression of activated caspase-1 in primary cultured rat cortical astrocytes and bilirubin 6 h after intervention was significantly higher than that in control group (P <0.05). Compared with bilirubin group, VX-765 treatment inhibited (P <0.05), and increased the cell survival rate (P <0.05), and decreased the release of IL-1β in the culture supernatant (P <0.01). CONCLUSION: Bilirubin can induce an increase in the expression of activated caspase-1. Inhibition of caspase-1 activation by VX-765 can reduce the neurotoxicity of bilirubin, enhance the survival rate of primary cultured astrocytes and reduce the release of inflammatory cytokines.