Nrf2/NLRP3信号通路在丙泊酚后处理减轻氧糖剥夺-复糖复氧大鼠海马神经元损伤中的作用  

Role of Nrf2/NLRP3 signaling pathway in propofol postconditioning-induced reduction of hippocampal neuron injury in a rat model of oxygen-glucose deprivation and restoration

作　　者：朱敏[1,2] 于洪丽[2] 孙英[2] 李红霞[2] 任恒昌[2] 喻文立[2] Zhu Min;Yu Hongli;Sun Ying;Li Hongxia;Ren Hengchang;Yu Wenli(First Central Clinical School,Tianjin Medical University,Tianjin 300192,China;Department of Anesthesiology,Tianjin First Central Hospital,Tianjin 300192,China)

机构地区：[1]天津医科大学一中心临床学院,天津300192 [2]天津市第一中心医院麻醉科,天津300192

出　　处：《中华麻醉学杂志》2023年第8期977-980,共4页Chinese Journal of Anesthesiology

基　　金：天津市医学重点学科(专科)建设项目;国家自然科学基金面上项目(82092219);天津市第一中心医院院级春风项目(院2020CF04)。

摘　　要：目的:评价核因子E2相关因子2/核苷酸结合寡聚化结构域样受体蛋白3(Nrf2/NLRP3)信号通路在丙泊酚后处理减轻氧糖剥夺-复糖复氧大鼠海马神经元损伤中的作用。方法:原代培养孕16~18 d Wistar大鼠胎鼠海马神经元7 d,采用随机数字表法分为4组(n=42):对照组(C组)正常培养;氧糖剥夺-复糖复氧组(O组)氧糖剥夺1 h后复糖复氧;丙泊酚后处理组(P组)复糖复氧即刻加入丙泊酚(终浓度1.2μg/ml)孵育2 h,更换为正常培养液;Nrf2 siRNA(-)转染组(N组)原代神经元培养第3天行携带Nrf2基因敲除的慢病毒转染,24 h后更换为正常培养液,第7天进行模型制备及丙泊酚后处理。于继续培养24 h时收集细胞,采用流式细胞术检测神经元凋亡率,RT-PCR法检测Nrf2和NLRP3 mRNA表达,Western blot法检测Nrf2和NLRP3表达,ELISA法测定TNF-α、IL-6和IL-1β浓度;试剂盒法检测还原性谷胱甘肽(GSH)、SOD和过氧化氢酶(CAT)活性。结果:与C组比较,O组和P组神经元凋亡率升高,TNF-α、IL-6和IL-1β浓度升高,GSH、SOD和CAT水平降低,Nrf2、NLRP3及其mRNA表达上调,Nrf2细胞核/浆比例增加(P<0.05);与O组比较,P组神经元凋亡率降低,TNF-α、IL-6和IL-1β浓度降低,GSH、SOD和CAT水平升高,Nrf2及其mRNA表达上调,Nrf2细胞核/浆比例增加,NLRP3及其mRNA表达下调(P<0.05);与P组比较,N组神经元凋亡率升高,TNF-α、IL-6和IL-1β浓度升高,GSH、SOD和CAT水平降低,Nrf2及其mRNA表达下调,Nrf2细胞核/浆比例降低,NLRP3及其mRNA表达上调(P<0.05)。结论:Nrf2/NLRP3信号通路参与了丙泊酚后处理减轻氧糖剥夺-复糖复氧大鼠海马神经元损伤的过程。Objective To evaluate the role of nuclear factor erythroid 2-related factor 2(Nrf2)/nucleotide-binding oligomerization domain-like receptor family pyrin domain containing 3(NLRP3)signaling pathway in propofol postconditioning-induced reduction of hippocampal neuron injury in a rat model of oxygen-glucose deprivation and restoration(OGD/R).Methods The hippocampal neurons were isolated from fetal rats of Wistar rats at 16-18 days of gestation and primarily cultured for 7 days and then divided into 4 groups(n=42 each)using a random number table method:control group(group C),OGD/R group(group O),propofol post-conditioning group(group P)and Nrf2 siRNA(-)transfection group(group N).The cells were routinely cultured in group C.The cells were subjected to oxygen-glucose deprivation for 1 h followed by oxygen and glucose supply in group O.Propofol(final concentration 1.2μg/ml)was added immediately after oxygen and glucose supply,the cells were then cultured for 2 h,and the culture medium was replaced with the normal culture medium in group P.The primarily cultured neurons were transfected with Nrf2 gene knockout lentivirus on 3rd day of culture,24 h later the cells were then routinely cultured,and the model was prepared and propofol conditioning was performed on 7th day.Cells were collected at 24 h of incubation for determination of the cell apoptosis(by flow cytometry),expression of Nrf2 and NLRP3 mRNA and protein(using quantitative real-time polymerase chain reaction or Western blot),concentrations of tumor necrosis factor-alpha(TNF-α),interleukin-6(IL-6)and IL-1β,and activities of glutathione(GSH),superoxide dismutase(SOD)and catalase(CAT)(kit method).Results Compared with group C,the apoptosis rate of neurons was significantly increased,concentrations of TNF-α,IL-6 and IL-1βwere increased,the levels of GSH,SOD and CAT_(3)were decreased,the expression of Nrf2 and NLRP3 protein and mRNA was up-regulated,and the nuclear/plasma ratio of Nrf2 was increased in O and P groups(P<0.05).Compared with group O,the apoptosis

关 键 词：NF-E2相关因子2 NLR家族 热蛋白结构域包含蛋白3 二异丙酚 缺血后处理 海马 神经元 低氧 

分 类 号：R614[医药卫生—麻醉学]