皮层星形胶质细胞Toll样受体4在炎症和β淀粉样蛋白形成中的作用（英文）  

The role of Toll-like receptor 4 on inflammation and Aβ formation in cortex astrocytes

作　　者：龚长银 周爱玲[1] 茅家慧[1] 胡亚娥[1] 耿劲松[3] 

机构地区：[1]南通大学医学院病理生理学系,南通226001 [2]南京医科大学附属常州市第二人民医院病理科,常州213003 [3]南通大学医学院循证医学中心,南通226001

出　　处：《生理学报》2014年第6期631-638,共8页Acta Physiologica Sinica

基　　金：supported by the Applied Research and Technology Program of Nantong Municipality,Jiangsu Province,China(No.BK2013007);the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions(PAPD)(2011),Jiangsu Province,China

摘　　要：为了探讨星形胶质细胞在炎症和β淀粉样蛋白(amyloidβ-protein,Aβ)形成中的作用和可能的分子机制,本研究通过LPS刺激体外培养大鼠皮层星形胶质细胞,首先采用实时PCR和Western blot方法分别检测Toll样受体4(Toll-like receptor 4,TLR4)、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)、白细胞介素-1β(interleukin 1β,IL-1β)、β淀粉样前体蛋白(β-amyloid precursor protein,β-APP)和β-位点APP剪切酶1(β-site APP clearing enzyme 1,BACE1)m RNA及TLR4、NF-κB/P65蛋白水平,而后用免疫荧光法进一步证明NF-κB/P65的核易位,ELISA法测定培养上清中TNF-α、IL-1β和Aβ含量。结果显示,这些指标在LPS刺激后均不同程度地上调。然而,如果预先用TLR4抗体处理,与仅用LPS刺激组相比,LPS对NF-κB/P65核易位及培养上清中TNF-α、IL-1β和Aβ含量的刺激作用显著减弱或消失。结果表明,星形胶质细胞TLR4可能通过TLR4/NF-κB信号通路在炎症和Aβ的形成中发挥重要的作用。To investigate the role and possible molecular mechanism of astrocytes in inflammation and amyloid β-protein（Aβ） formation, in this research, by using LPS to stimulate cultured rat astrocytes in vitro with or without anti-Toll-like receptor 4（TLR4） antibody pretreatment, we first detected the TLR4, TNF-α, IL-1β, β-amyloid precursor protein（β-APP） and β-site APP clearing enzyme 1（BACE1） m RNA with real-time PCR, and TLR4, NF-κB/P65 protein in cultured astrocytes by Western blot, and then further probed the translocation of NF-κB/P65 using immunofluorescence and the contents of TNF-α, IL-1β and Aβ in culture supernatant through ELISA. We found that all of these indexes increased at different degrees after LPS-stimulation. However, if pretreatment with antiTLR4 antibody, such stimulating effects of LPS on the nuclear translocation of NF-κB/P65 and TNF-α, IL-1β, Aβ contents in astrocytic culture supernatant were reduced significantly or disappeared in comparison with the group with only LPS-administration. Our results suggest that TLR4 in astrocytes might play an important role in the inflammation and Aβ formation through the TLR4/NF-κB signaling pathway, thus providing new knowledge and understanding of the inflammatory hypothesis of AD pathogenesis.

关 键 词：阿尔茨海默病 星形胶质细胞 TOLL样受体4 炎症 Β淀粉样蛋白 

分 类 号：R363[医药卫生—病理学]