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    刘芷含, 陈巍巍, 岳婵娟, 吴明凤, 滕雪, 陈锐, 杨龙, 杜波, 程言博. Nod样受体蛋白3(NLRP3)在大鼠原代皮质神经元坏死样凋亡中的表达[J]. 徐州医科大学学报, 2017, 37(5): 293-297.
    引用本文: 刘芷含, 陈巍巍, 岳婵娟, 吴明凤, 滕雪, 陈锐, 杨龙, 杜波, 程言博. Nod样受体蛋白3(NLRP3)在大鼠原代皮质神经元坏死样凋亡中的表达[J]. 徐州医科大学学报, 2017, 37(5): 293-297.
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    LIU Zhihan, CHEN Weiwei, YUE Chanjuan, WU Mingfeng, TENG Xue, CHEN Rui, YANG Long, DU Bo, CHENG Yanbo. The expression of Nod-like receptor protein 3 (NLRP3) in necroptosis of primary cortical neurons[J]. Journal of Xuzhou Medical University, 2017, 37(5): 293-297.
    Citation: LIU Zhihan, CHEN Weiwei, YUE Chanjuan, WU Mingfeng, TENG Xue, CHEN Rui, YANG Long, DU Bo, CHENG Yanbo. The expression of Nod-like receptor protein 3 (NLRP3) in necroptosis of primary cortical neurons[J]. Journal of Xuzhou Medical University, 2017, 37(5): 293-297.
    shu

    Nod样受体蛋白3(NLRP3)在大鼠原代皮质神经元坏死样凋亡中的表达

    The expression of Nod-like receptor protein 3 (NLRP3) in necroptosis of primary cortical neurons

      摘要
    • 摘要: 目的探讨Nod样受体蛋白3 (NLRP3)在大鼠原代皮质神经元坏死样凋亡中的表达情况。方法SD大鼠原代皮质神经元细胞培养12天，Q-VD-Oph预处理30 min后缺糖缺氧刺激诱导坏死样凋亡。①将细胞分为正常对照组、缺糖缺氧组（缺糖缺氧2 h，复氧12 h）、实验组（缺糖缺氧2 h，复氧3、6、12、24 h，均用Q-VD-Oph处理）,Western blot检测NLRP3蛋白表达，生化分析仪检测乳酸脱氢酶（LDH）水平；②将细胞分为正常对照组、缺糖缺氧组（缺糖缺氧2 h，复氧12 h）、实验组（缺糖缺氧0.5、1、1.5、2、3 h，复氧12 h，均用Q-VD-Oph处理）,再次进行NLRP3及LDH测定；③细胞缺糖缺氧2 h、复氧12 h（用Q-VD-Oph处理）后免疫荧光观察NLRP3的表达。结果缺糖缺氧2 h后，随着复氧时间的延长，NLRP3在12 h表达量最高，24 h表达量较12 h下降（P<0.05），LDH在12 h的表达量最高。缺糖缺氧时间不同，各组均复氧12 h时，NLRP3在缺糖缺氧 2 h表达量最高，3 h蛋白表达量较2 h下降（P<0.05），LDH在2 h的表达量最高。在缺糖缺氧2 h、复氧12 h，细胞免疫荧光强度最高。结论NLRP3在原代皮质神经元细胞坏死样凋亡中有表达，且随着NLRP3表达量升高，细胞受损程度相应增高。

       

      Abstract: ObjectiveTo investigate the levels of Nod-like receptor protein 3 (NLRP3) in primary cortical neurons with necroptosis. MethodsPrimary cortical neurons of SD rats were cultured for 12 days, before pretreated with Q-VD-Oph for 30min followed by oxygen-glucose deprivation to induce necroptosis. ① The cells were divided into the following groups: a normal control group, an oxygen-glucose deprivation group (oxygen-glucose deprivation for 2 h followed by reperfusion for 12 h), and treatment groups (oxygen-glucose deprivation for 2 h followed by reperfusion for 3, 6, 12 and 24 h, and all of them were treated with Q-VD-Oph) Their levels of NLRP3 and lactate dehydrogenase (LDH) were examined. ② The cells were divided into the following groups: a control group, an oxygen-glucose deprivation group (oxygen-glucose deprivation for 2 h followed by reperfusion for 12 h), and treatment groups (oxygen-glucose deprivation for 0.5, 1, 1.5, 2, and 3 h followed by reperfusion for 12 h, and all of them were treated with Q-VD-Oph). Their levels of NLRP3 and LDH were examined. ③ The cells were subjected to oxygen-glucose deprivation for 2 h followed by reperfusion for 12 h (all of them were treated with Q-VD-Oph) before determination of their levels of NLRP3 by immunofluorescence. ResultsAfter oxygen-glucose deprivation for 2 h, the level of NLRP3 was increased as the time of reperfusion extended, which then reached the highest at 12 h and was remarkably decreased at 24 h (P<0.05). The level of LDH reached the highest at 12 h. After oxygen-glucose deprivation for different periods of time followed by reperfusion for 12 h, the level of NLRP3 reached the highest when oxygen-glucose deprivation for 2 h, which then was remarkably decreased at 3 h (P<0.05). The level of LDH reached the highest when oxygen-glucose deprivation for 2 h. The immunofluorescence intensity was the highest when the cells were oxygen-glucose deprived for 2 h followed by reperfusion for 12 h. ConclusionsNLRP3 is present in primary cortical neuron with necroptosis. The degree of cell injury is gradually increased ass the level of NLRP3 increases.

       

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