南方医科大学学报

南方医科大学学报 ›› 2018, Vol. 38 ›› Issue (05): 567-.

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Caspase-1活化在胆红素诱导的大鼠海马神经元损伤中的作用

韦倩，冯洁，何春梅，华子瑜   

  • 出版日期:2018-05-20 发布日期:2018-05-20

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

  • Online:2018-05-20 Published:2018-05-20

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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（50 μmol/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（50 μmol/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。

Abstract: 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 after 50 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.