摘要
[^(35)S]-甲硫氨酸掺入法测定细胞Aβ分泌速率,观察知母活性成分ZMS和黄芪活性成分AST和HT,及两药合用对Aβ生成速率的影响。用[35S]-甲硫氨酸标记转染APP695基因的细胞,所获细胞上清以Aβ22-35单克隆抗体进行免疫共沉淀,结合Western印迹法,以配对对照(加载体)的Aβ条带灰度值为1,计算各用药组灰度的相对强度。本方法能稳定地测到细胞在24—72h内分泌的Aβ量,同一受试样品在不同批实验中的变异(CV)在17%—32%之间。在培养液中中药活性成分的浓度为1×10?5mol/L时,受试物质中AST及ZMS+AST合用对Aβ的分泌速率有明显抑制作用(P<0.01)。另两种受试物则无作用。黄芪活性成分AST有抑制Aβ分泌的作用,ZMS与AST两药合用作用更为显著。
To observe the effect of certain active components from traditional Chinese medicinal herbs on Aβ secretion rates with L-[^35S]-Methionine, β-amyloid peptide （Aβ in SK-N-SH cell lines stably transfected with APP695 was metabolically labeled with L-[^35S]-Methionine. The supematant from culture medium was immunoprecipitated with monoclonal antibody against Aβ22-35, Western blot was carded out, and the gray density of Aβ band in the autoradiograph was measured by an image analysis system. The active components from certain traditional Chinese medicinal herbs （ZMS from Zhimu and AST and HT from Huangqi） were added to the culture medium at a final concentration of 10^-5 mol/L. An Aβ band in the autoradiograph was clearly viewed in the culture medium after 24 h incorporation of [^35S]-Methionine which represent the secretion rate of Aβ by the cells. One of the 3 tested components （AST） could significantly reduce the AI3 secretion rate while the other two showed no effect. The preliminary result showed that certain active component from traditional Chinese medicines could decrease the Aβ secretion rate but other active components could not. Combined use of the AST and ZMS was more effective than single AST.
出处
《核技术》
EI
CAS
CSCD
北大核心
2006年第10期773-776,共4页
Nuclear Techniques
基金
国家自然科学基金项目(30070926)资助
