文章摘要
卢晓星,张小强,苗歆雨,季倩倩,李 琪,陶一凡.原花青素对LPS/ATP诱导的NLRP3炎症小体激活及NF⁃κBp65磷酸化的抑制作用[J].南京医科大学学报,2020,(8):1111~1118
原花青素对LPS/ATP诱导的NLRP3炎症小体激活及NF⁃κBp65磷酸化的抑制作用
Inhibitory effects of proanthocyanidin on LPS/ATP induced activation of NLRP3 inflammasome and phosphorylation of NF⁃κBp65
投稿时间:2019-12-25  
DOI:10.7655/NYDXBNS20200806
中文关键词: 原花青素  炎症因子  NLRP3炎症小体  脂多糖  腺嘌呤核苷三磷酸
英文关键词: proanthocyanidin  inflammatory factors  NLRP3 inflammasome  lipopolysaccharide  adenine nucleoside triphosphate
基金项目:中央高校基本科研业务费专项资金;江苏省普通高校研究生科研创新计划(SJZZ16_0034)
作者单位
卢晓星 东南大学公共卫生学院环境医学工程教育部重点实验室江苏 南京 210009 
张小强 东南大学公共卫生学院环境医学工程教育部重点实验室江苏 南京 210009 
苗歆雨 东南大学公共卫生学院环境医学工程教育部重点实验室江苏 南京 210009 
季倩倩 东南大学公共卫生学院环境医学工程教育部重点实验室江苏 南京 210009 
李 琪 东南大学公共卫生学院环境医学工程教育部重点实验室江苏 南京 210009 
陶一凡 东南大学公共卫生学院环境医学工程教育部重点实验室江苏 南京 210009 
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中文摘要:
      目的:研究原花青素对脂多糖(lipopolysaccharide,LPS)与腺嘌呤核苷三磷酸(adenosine triphosphate,ATP)联合诱导的小鼠小胶质细胞BV2 NOD样受体蛋白3(nucleotide?binding oligomerization domain?like receptor protein 3,NLRP3)炎症小体激活及核因子(nuclear factor,NF)?κBp65磷酸化的抑制作用。方法:以1.0 μg/mL LPS、2.5 μmol/L ATP诱导BV2细胞炎症损伤模型,用不同浓度原花青素(0.1、1.0、2.5、5.0 μg/mL)干预细胞。采用噻唑蓝(thiazolyl blue tetrazolium bromide,MTT)法检测细胞存活率;比色法检测一氧化氮(nitric oxide,NO)释放水平;微量酶标法测乳酸脱氢酶(lactate dehydrogenase,LDH)活力;ELISA法检测白细胞介素(interleukin,IL)?1β、IL?18的分泌水平;Western blot法检测NLRP3、凋亡相关斑点样蛋白(apoptosis?associated speck?like protein containing a CARD,ASC)、半胱氨酸天冬氨酸蛋白酶(caspase)?1、pro?caspase?1、p?NF?κBp65、NF?κBp65蛋白表达水平。结果:不同浓度的原花青素对BV2细胞存活率的影响与对照组相比差异无统计学意义(P > 0.05)。与对照组相比,LPS/ATP诱导可增加BV2细胞NO、IL?1β、IL?18水平以及LDH活力(P < 0.05),增加NLRP3、ASC、pro?caspase?1、caspase?1、
英文摘要:
      Objective:To observe the inhibitory effects of proanthocyanidin on activation of nucleotide?binding oligomerization domain?like receptor protein 3(NLRP3)inflammasome and phosphorylation of nuclear factor(NF)?κBp65 induced by lipopolysaccharide(LPS)and adenine nucleoside triphosphate(ATP)in mouse microglia(BV2). Methods:BV2 cells were stimulated with 1.0 μg/mL LPS and 2.5 μmol/L ATP,and treated with different concentrations of proanthocyanidin(0.1,1.0,2.5,5.0 μg/mL). Cell viability was determined by thiazolyl blue tetrazolium bromide(MTT)assay. Nitric oxide(NO)release was detected by colorimetry. The activity of lactate dehydrogenase(LDH)was measured by microenzyme labeling method. The secretion of interleukin(IL)?1β and IL?18 were determined by ELISA. Expression of NLRP3,apoptosis?associated speck?like protein containing a CARD(ASC),caspase?1,pro?caspase?1,p?NF?κBp65,NF?κBp65 were detected by Western blot. Results:The effect of different concentrations of proanthocyanidin on the survival rate of BV2 cells was not statistically significant compared with the control group(P > 0.05). Compared with the control group,LPS/ATP increased the secretions of NO,IL?1β,IL?18 and LDH activity(P < 0.05),and the expressions of NLRP3,ASC,pro?caspase?1,caspase?1,p?NF?κBp65(P < 0.05). Compared with the LPS/ATP group,proanthocyanidin reduced the secretions of NO,IL?1β,IL?18 and LDH activity of BV2 cells(P < 0.05). In addition,proanthocyanidin(1.0,2.5,5.0 μg/mL) decreased the expressions of NLRP3,ASC,pro?caspase?1 and caspase?1(P < 0.05). Similarly,NF?κB inhibitor BAY11?7082(5.0 μmol/L)reduced NF?κBp65 phosphorylation and the expressions of NLRP3,ASC,pro?caspase?1,and caspase?1(P < 0.05). Conclusion:Proanthocyanidin can inhibit secretion of inflammatory factor and activation of NLRP3 inflammasome induced by LPS/ATP,which is closely related to the inhibition of phosphorylation of NF?κBp65 by proanthocyanidin in LPS/ATP induced status.
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