[1]刘芷含,陈巍巍,岳婵娟,等. 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(05):293-297.
 LIU Zhihan,CHEN Weiwei,YUE Chanjuan,et al. The expression of Nod-like receptor protein 3 (NLRP3) in necroptosis of primary cortical neurons[J].Journal of Xuzhou Medical University,2017,37(05):293-297.
点击复制

 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()
分享到:

《徐州医科大学学报》[ISSN:2096-3882/CN:32-1875/R]

卷:
37
期数:
2017年05期
页码:
293-297
栏目:
出版日期:
2017-05-25

文章信息/Info

Title:
 The expression of Nod-like receptor protein 3 (NLRP3) in necroptosis of primary cortical neurons
作者:
 刘芷含1陈巍巍2岳婵娟3吴明凤1滕雪1陈锐1杨龙4杜波4程言博4*
 1.徐州医科大学研究生学院,江苏徐州221004;2.徐州市中心医院神经内科,江苏徐州221009;
3.徐州市中心医院检验科;4.徐州医科大学附属医院神经内科,江苏徐州221002
Author(s):
 LIU Zhihan1 CHEN Weiwei2 YUE Chanjuan3 WU Mingfeng1 TENG Xue1 CHEN Rui1 YANG Long4 DU Bo4 CHENG Yanbo4*
 1.Graduate School, Xuzhou Medical University, Xuzhou, Jiangsu 221004, China;
2.Department of Neurology, Xuzhou Central Hospital, Xuzhou, Jiangsu 221009;
3.Department of Clinical laboratory, Xuzhou Central Hospital; 4.Department of Neurology,
the Affiliated Hospital of Xuzhou Medical University, Xuzhou, Jiangsu 221004,China
关键词:
 Nod样受体蛋白3坏死样凋亡
Keywords:
 Nod-like receptor protein 3 necroptosis
分类号:
R743.33
文献标志码:
A
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.

参考文献/References:

 [1]Broughton BR, Reutens DC, Sobey CG. Apoptotic mechanisms after cerebral ischemia [J]. Stroke, 2009,40(5):e331-e339.
[2]Sims NR, Muyderman H. Mitochondria, oxidative metabolism and cell death in stroke [J]. Biochim Biophys Acta, 2010,1802(1):80-91.
[3]Yang F, Wang Z, Wei X, et al. NLRP3 deficiency ameliorates neurovascular damage in experimental ischemic stroke [J]. J Cereb Blood Flow Metab, 2014,34(4):660-667.
[4]Heneka MT, Kummer MP, Stutz A, et al. NLRP3 is activated in Alzheimer’s disease and contributes to pathology in APP/PS1 mice [J]. Nature, 2013,493(7434):674-678.
[5]柏华,赵学军,张启芳,等.钙离子强化NLRP3炎症小体引起的神经母细胞瘤细胞氧化应激研究[J]. 中华行为医学与脑科学杂志,2016,25(3):210-214.
[6]Dalmas E, Venteclef N, Caer C, et al. T cell-derived IL-22 amplifies IL-1β-driven inflammation in human adipose tissue: relevance to obesity and type 2 diabetes [J]. Diabetes, 2014,63(6):1966-1977.
[7]Mehta SL, Manhas N, Raghubir R. Molecular targets in cerebral ischemia for developing novel therapeutics [J]. Brain Res Rev, 2007,54(1):34-66.
[8]Mulay SR, Desai J, Kumar SV, et al. Cytotoxicity of crystals involves RIPK3-MLKL-mediated necroptosis [J]. Nat Commun, 2016,7:10274.
[9]Moujalled DM, Cook WD, Murphy JM, et al. Necroptosis induced by RIPK3 requires MLKL but not Drp1 [J]. Cell Death Dis, 2014,5:e1086.
[10]Rosenbaum DM, Degterev A, David J, et al. Necroptosis, a novel form of caspase-independent cell death, contributes to neuronal damage in a retinal ischemia-reperfusion injury model [J]. J Neurosci Res, 2010,88(7):1569-1576.
[11]Fann DY, Lee SY, Manzanero S, et al. Intravenous immunoglobulin suppresses NLRP1 and NLRP3 inflammasome-mediated neuronal death in ischemic stroke [J]. Cell Death Dis, 2013,4:e790.
[12]冯涛,丁红梅,耿德勤.去泛素化酶在皮质神经元缺糖缺氧再灌注损伤后坏死样凋亡中的表达[J].中华行为医学与脑科学杂志,2013,22(8):692-695.
[13]陈巍巍,张翠翠,程言博,等.受体相互作用蛋白3在大鼠皮质神经元坏死性凋亡信号通路中的作用[J].中华行为医学与脑科学杂志,2011,20(6):481-484.
[14]Bauernfeind F, Rieger A, Schildberg FA, et al. NLRP3 inflammasome activity is negatively controlled by miR-223 [J]. J Immunol, 2012,189(8):4175-4181.
[15]Zhou R, Yazdi AS, Menu P, et al. A role for mitochondria in NLRP3 inflammasome activation [J].Nature, 2011,469(7329):221-225.
[16]Duewell P, Kono H, Rayner KJ, et al. NLRP3 inflammasomes are required for atherogenesis and activated by cholesterol crystals [J]. Nature, 2010,464(7293):1357-1361.
[17]Mariathasan S, Weiss DS, Newton K, et al. Cryopyrin activates the inflammasome in response to toxins and ATP [J].Nature, 2006,440(7081):228-232.
[18]Rathinam VA, Vanaja SK, Fitzgerald KA. Regulation of inflammasome signaling [J].Nat Immunol, 2012,13(4):333-342.
收稿日期:2017-03-10修回日期:2017-05-10

相似文献/References:

[1]孙秀媛,李海玲,朱相华,等.rTMS及MECT在不同时间点对抑郁症疗效和BDNF的影响及相关性研究[J].徐州医科大学学报,2017,37(04):262.
[2]代春潇,宋远见,张云山,等. 大鼠MKK7真核表达质粒的构建及其在COS7细胞中转染效率[J].徐州医科大学学报,2017,37(04):211.
  [J].Journal of Xuzhou Medical University,2017,37(05):211.
[3]张端强,丁晓帆,杨丹丹,等. 加综合征患者复发影响因素研究[J].徐州医科大学学报,2017,37(04):216.
 [J].Journal of Xuzhou Medical University,2017,37(05):216.
[4]王立新,袁峰*,郭开今,等. 纳米羟基磷灰石/细菌纤维素对兔骨髓间充质干细胞黏附及成骨分化的影响[J].徐州医科大学学报,2017,37(04):221.
 [J].Journal of Xuzhou Medical University,2017,37(05):221.
[5]李海青,曹猛,江涛,等. PD-L1在结直肠癌中的表达及其临床意义[J].徐州医科大学学报,2017,37(04):227.
 [J].Journal of Xuzhou Medical University,2017,37(05):227.
[6]刘兵,唐明生,李苹,等. 用随机森林模型分析胃癌高危人群干预效果的影响因素*[J].徐州医科大学学报,2017,37(04):231.
 [J].Journal of Xuzhou Medical University,2017,37(05):231.
[7]周新宇?,牛坚,曹宽,等. Tim3在炎症反应调控及对肝细胞损伤保护机制研究[J].徐州医科大学学报,2017,37(04):236.
 [J].Journal of Xuzhou Medical University,2017,37(05):236.
[8]商书霞,宋光耀*,刘晓耕,等. 血清胆红素和γ-谷氨酰转肽酶与糖尿病患者非酒精性脂肪肝的相关性研究[J].徐州医科大学学报,2017,37(04):245.
 [J].Journal of Xuzhou Medical University,2017,37(05):245.
[9]李新文1,巩尊科2,王世雁2,等. RS在非痴呆型血管性认知障碍评定中的临床应用研究[J].徐州医科大学学报,2017,37(04):248.
 [J].Journal of Xuzhou Medical University,2017,37(05):248.
[10]樊叶,韩维维,何帮顺,等.替格瑞洛含药血清对人胃黏膜上皮细胞GES-1的损伤作用[J].徐州医科大学学报,2017,37(04):252.
 [J].Journal of Xuzhou Medical University,2017,37(05):252.

备注/Memo

备注/Memo:
 基金项目:国家自然科学基金(81501138);徐州市科技项目(KC15SH077)
*通信作者,E-mail:cyanbo@126.com

收稿日期:2017-03-10修回日期:2017-05-10
更新日期/Last Update: 2017-06-09