摘要
目的研究α_1 抗糜蛋白酶(ACT)与类淀粉样蛋白Aβ_(1-42)在体外形成复合物的性质,以及该复合物对神经母细胞瘤细胞(Kelly)表达核转录因子过氧化物酶体增殖子激活受体γ(PPARγ)和核因子κB(NFκB)的影响。方法Aβ_(1-42)与ACT溶液以10∶1mol/L混合,用琼脂糖凝胶电泳检测2h和24h反应产物Aβ_(1-42)/ACT(2h)、Aβ_(1-42)/ACT(24h),电泳迁移率检测法(EMSA)检测Aβ_(1-42)/ACT复合物对Kelly细胞表达PPARγ和NFκB的影响。结果Aβ_(1-42)与ACT分子之间可以结合形成复合物,而且复合物的功能随反应时间的不同而不同与对照组细胞相比,Aβ_(1-42)/ACT(2h)使Kelly细胞对PPARγ和NFκB的表达分别增加158% (P<0 05)和77% (P<0 05),而Aβ_(1-42)/ACT(24h)和Aβ_(1-42)、ACT单个分子则无类似作用。结论PPARγ和NFκB是调节人体炎性反应过程的重要细胞核转录因子,与阿尔茨海默病(AD)的病理改变相关。ACT通过与Aβ_(1-42)结合影响两种细胞因子的表达,可能是AD中神经系统炎性反应平衡失调的病理通路之一,提示对AD的发病有影响。
Objective To study the interaction between α_1-antichymotrypsin (ACT) and amyloid peptide Aβ_~1-42 in vitro, and the effects of their complex on expression of transcription factors PPARγ and NFκB in human neuroblastoma (Kelly) cells. Methods Aβ_~1-42 and ACT were mixed at a 10∶1 molar ratio at room temperature, the complex formed in 2 hours and 24 hours were expressed as Aβ_~1-42 /ACT (2 hours), Aβ_~1-42 /ACT (24 hours), respectively. The complexes were studied by using agrose electrophoresis, and their effects on PPARγ and NFκB expressions were assessed by electrophoresis mobility shift assay (EMSA). Results At conditions used, ACT interacted with Aβ_~1-42 in vitro, the complexes formed and functioned differently according to their interaction period. Compared with unstimulated control cells, Aβ_~1-42 /ACT (2 hours) strongly increased PPARγ and NFκB expressions by Kelly cells by 158% (P<0.05) and 77% (P<0.05), respectively, while Aβ_~1-42 /ACT (24 hours), or Aβ_~1-42 , ACT alone had no similar effects. Conclusions PPARγ and NFκB should be two transcription factors actively involved in the inflammatory process in human bodies. They have also been found increased in Alzheimer’s brains, suggesting they should be a factor of pathology in this disease. The interaction between ACT and Aβ_~1-42 had an effect on the expression of PPARγ and NFκB, which might be responsible for the imbalanced inflammation in the central nervous system in Alzheimer’s disease (AD). The relative status and quantity of ACT and Aβ1-42 in the amyloid deposits might have an impact on the pathogenesis of AD.
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2005年第2期82-86,共5页
Chinese Journal of Neurology
关键词
Α1-抗糜蛋白酶
类淀粉样蛋白Aβ1-42
细胞表达
核转录因子
阿尔茨海默病
Alzheimer disease
alpha-antichymotrypsin
Amyloid beta protein
Peptide fragments
Receptors,cytoplasmic and nuclear
Transcription factors
NF-kappa B
