黄连解毒汤对Aβ25-35诱导的HT22细胞NF-κB活化及炎症因子水平的影响  

Effect of Huanglian Jiedutang on Aβ25-35-induced Activation of NF-κB and Level of Inflammatory Factor in HT22 Cells

作　　者：张茹兰 黄秀芳 陶彦谷 李建军[1] 黄启辉[1] ZHANG Ru-lan;HUANG Xiu-fang;TAO Yan-gu;LI Jian-jun;HUANG Qi-hui(Sun Yat-Sen memory hospital of Sun Yat-sen University, Guangzhou 510300, China;Guangzhou University of Chinese Medicine, Guangzhou 510080, China)

机构地区：[1]中山大学孙逸仙纪念医院,广州510300 [2]广州中医药大学,广州510080

出　　处：《中国实验方剂学杂志》2018年第13期159-165,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：广东省中医药局科研项目(20171065)

摘　　要：目的：探讨黄连解毒汤对β-淀粉样蛋白25-35（β-amyloid protein 25-35,Aβ（25-35））诱导的HT22细胞核转录因子-κB（NF-κB）活化及炎症因子白细胞介素-1β（IL-1β）,IL-6,肿瘤坏死因子-α（TNF-α）的影响。方法：培养HT22细胞,HT22细胞分为空白组、模型组、多奈哌齐组（0.000 9 g·kg-1）和黄连解毒汤低、高剂量组（2.7,8.1 g·kg-1）,空白组、模型组加入空白血清,各药物组分别加入含药血清1 h后,模型组和各药物组细胞分别加入Aβ（25-35）（40μmol·L-1）,24 h后运用蛋白免疫印迹法（Western blot）检测p-NF-κB p65表达水平;免疫荧光法检测分析各组NF-κB p65核移位情况;用酶联免疫吸附测定（ELISA）检测IL-1β,IL-6,TNF-α蛋白含量及实时荧光定量聚合酶链式反应（Real-time PCR）检测IL-1β,IL-6,TNF-αmRNA的表达情况。结果：与空白组比较,模型组的IL-1β,IL-6,TNF-α蛋白分泌及其mRNA表达增多,NF-κB活化水平增高（P〈0.05）;与模型组比较,黄连解毒汤低、高剂量组及多奈哌齐组的炎性因子及mRNA表达量均减低（P〈0.05）,并且能抑制NF-κB活化。结论：黄连解毒汤可降低Aβ（25-35）诱导HT22细胞炎症反应,从而减轻Aβ（25-35）神经毒性作用,作用机制可能与抑制NF-κB活化有关。Objective： To investigate the effect of Huanglian Jiedutang on the activation of nuclear transcription factor κB（ NF-κB） and the level of inflammatory factors including interleukin-1β（ IL-1β）,IL-6 and tumor necrosis factor-α（ TNF-α） of HT22 cells induced by β-amyloid protein 25-35（ Aβ（25-35））. Method： The HT22 cells were cultured and divided into five groups： blank group,model group,donepezil-treated group（ 0. 000 9 g·kg-1） and low and high doses Huanglian Jiedutang-treated groups（ 2. 7,8. 1 g·kg-1）. The HT22 cells in blank group and model group were treated with blank serum and various treatment groups were treated with corresponding drug-serum and cultured for 1 h,following the incubation with 40 μmol·L-1 Aβ（25-35） for 24 h in model group and treatment groups. Then the protein expression level of p-NF-κB p65 was detected by Western blot.Nuclear translocation of NF-κB p65 was determined by immunofluorescence assay. The protein expression and mRNA expression levels of IL-1β,IL-6 and TNF-α were respectively detected by enzyme-linked immunosorbent assay（ ELISA） and real time fluorescence quantification polymerase chain reaction（ Reat-time PCR）. Result： As compared with the blank group,the protein and mRNA expression levels of IL-1 β,IL-6 and TNF-α and the activation level of NF-κB were increased in the model group（ P 〈0. 05）. As compared with the model group,low and high dose Huanglian Jiedutang and donepezil could decrease the inflammatory factors and mRNA expression levels（ P 〈0. 05） and inhibit NF-κB activation. Conclusion： Huanglian Jiedutang may reduce the Aβ（25-35）-induced HT22 cells inflammatory response and relieve Aβ（25-35） neurotoxicity,which may be related to inhibiting the activation of NF-κB.

关 键 词：阿尔茨海默病 Β淀粉样蛋白 黄连解毒汤 核转录因子-κB(NF-κB) 炎症因子 

分 类 号：R22[医药卫生—中医基础理论] R24[医药卫生—中医学]