持续乙醇摄入通过抑制NRF2通路促进HMGB1及其下游炎性因子表达  

Continuous alcohol intake promotes expression of HMGB1 and its downstream inflammatory factors by inhibiting NRF2 pathway

作　　者：李昕曈 孙光涛[1] 王梦 戚询中[1] 黄作义[1] 黄昕艳[2] 朱晓峰 LI Xintong;SUN Guangtao;WANG Meng;QI Xunzhong;HUANG Zuoyi;HUANG Xinyan;ZHU Xiaofeng(Department of Neurology,First Affiliated Hospital of Jiamusi University,Jiamusi,Heilongjiang 154002,China;Second Affiliated Hospital of Jiamusi University,Jiamusi,Heilongjiang 154002,China;Medical Research Center of Mudanjiang Medical College,Mudanjiang,Heilongjiang 157000,China)

机构地区：[1]佳木斯大学附属第一医院神经内科,黑龙江佳木斯154002 [2]佳木斯大学附属第二医院神经内科,黑龙江佳木斯154002 [3]牡丹江医学院医药研究中心,黑龙江牡丹江157000

出　　处：《重庆医学》2021年第14期2352-2356,共5页Chongqing medicine

基　　金：国家重点研发计划项目(2018YFC1314404);国家自然科学基金面上项目(81871041);黑龙江省教育厅基本科研业务费人才培养项目(2019-KYYWF-1357)。

摘　　要：目的分析核因子E2相关因子2(NRF2)在持续乙醇摄入调节小鼠前额叶高迁移率族蛋白B1(HMGB1)通路及炎性反应中的作用。方法腹腔注射乙醇构建持续乙醇摄入小鼠模型;并用乙醇干预原代神经元构建乙醇干预细胞模型;NRF2通路特异激动剂特丁基对苯二酚(tBHQ)对模型进行干预,选用实时荧光定量聚合酶链反应检测前额叶及神经元目的基因mRNA水平;选用Western blot检测蛋白表达变化。结果与对照组相比,乙醇干预组小鼠前额叶炎性因子肿瘤坏死因子-α(TNF-α)(P=0.015)、白细胞介素(IL)-1β(P=0.017)及HMGB1通路相关基因HMGB1(P=0.004)、TLR3(P=0.030)、TLR4(P=0.004)mRNA相对表达水平升高;NRF2(P=0.014、0.014)及HO-1(P=0.030、0.012)转录及蛋白表达水平均降低;与单纯乙醇摄入小鼠相比较,tBHQ干预后能够降低IL-1β(P=0.003)、HMGB1(P=0.000)、TLR4(P=0.015)mRNA水平及HMGB1(P=0.043)蛋白表达水平;在神经元实验中,与单纯乙醇干预组相比,tBHQ干预后神经元HMGB1(P=0.001、0.004)转录及蛋白水平明显下降。结论持续乙醇摄入通过抑制小鼠前额叶NRF2表达,进而激活HMGB1/TLR4通路,促进其下游IL-1β表达。Objective To analyze the role of NRF2 in the regulation of HMGB1 pathway and inflammatory response in the prefrontal lobe of mice by continuous alcohol intake.Methods The intraperitoneal injection of ethanol was used to establish the continuous alcohol intake model of mice;the primary neurons were intervened by ethanol to construct the ethanol interventional cellular model;the NRF2 pathway specific agonist conducted the intervention on the model.The mRNA level of target genes in the prefrontal lobe and neuron was detected by using the real time qRT-PCR.The change of protein expression was detected by Western blot.Results Compared with the control group,the relative expression levels of TNFα(P=0.015),IL-1β(P=0.017)and HMGB1 pathway related gene HMGB1(P=0.004),TLR3(P=0.030)and TLR4(P=0.004)mRNA in the mice prefrontal lobe of the ethanol intervention group were increased;the transcription and protein expression of NRF2(P=0.014,0.014)and HO-1(P=0.030,0.012)were decreased;compared with the simple ethanol intake mice,the tBHQ intervention could reduce the mRNA levels of IL-1β(P=0.003),HMGB1(P=0.000)and TLR4(P=0.015),and protein levels of HMGB1(P=0.043);in the neuron experiment,compared with the simple ethanol intervention group,the neuron HMGB1(P=0.001,0.004)transcription and protein levels after tBHQ intervention were significantly decreased.Conclusion Continuous ethanol intake can activate the HMGB1/TLR4 pathway,and promote its downstream IL-1βexpression by inhibiting the NRF2 expression in the prefrontal lobe of mice.

关 键 词：乙醇 前额叶 NRF2 HMGB1 炎性反应 

分 类 号：R749.62[医药卫生—神经病学与精神病学]