SYNTHETIC ALZHEIMER AMYLOID BETA-A4 PEPTIDES ENHANCE PRODUCTION OF COMPLEMENT C3 COMPONENT BY CULTURED MICROGLIAL CELLS
SYNTHETIC ALZHEIMER AMYLOID BETA-A4 PEPTIDES ENHANCE PRODUCTION OF COMPLEMENT C3 COMPONENT BY CULTURED MICROGLIAL CELLS
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DOI：
10.1016/0006-8993(93)91698-r
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发表时间：
1993-01-22
期刊：
影响因子：
2.9
通讯作者：
ISHII, T
中科院分区：
文献类型：
--
作者：
HAGA, S;IKEDA, K;ISHII, T
从新生小鼠制备的小胶质原代培养物显示出补体第三组分（C3）的产生和释放。新合成的[S-35]蛋氨酸标记C3用抗C3抗体免疫沉淀纯化。通过SDS-PAGE和荧光成像，从细胞裂解液中检测到C3为195 kDa条带，从培养上清中检测到C3为两个主要条带，对应于C3 α链（125 kDa）和β链（75 kDa），与已知的C3特征一致。内毒素脂多糖（LPS）促进了C3的生物合成。此外，C3的合成增加了5-10倍，以响应与淀粉样蛋白β /A4蛋白相对应的各种合成肽，淀粉样蛋白是阿尔茨海默病（AD）中细胞外淀粉样蛋白沉积的主要成分。在β /A4肽浓度从10马克杯/毫升到50马克杯/毫升范围内，C3合成的增加被证明是剂量依赖性的。这些结果表明，先前在淀粉样蛋白沉积物中发现的补体成分可能部分来源于与AD脑老年斑优先相关的反应性小胶质细胞。
Primary microglial cultures prepared from newborn mice showed the Production and release of the third component of complement (C3). Newly synthesized [S-35]methionine-labelled C3 was purified by immunoprecipitation using anti-C3-antibody. C3 was detected by SDS-PAGE and fluoroaraphy of the immunoprecipitated protein from cell lysates as a 195 kDa band, and from the supernatants of cultures as two major bands corresponding to the C3 alpha-chain (125 kDa) and beta-chain (75 kDa), consistent with known C3 characteristics. Increased biosynthesis of C3 was elicited by endotoxin lipopolysaccharide (LPS). Further, the synthesis of C3 was increased 5-10-fold in response to various synthetic peptides corresponding to the amyloid beta/A4 protein, which is the main constituent of extracellular amyloid deposits in Alzheimer's disease (AD). The increased synthesis of C3 was shown to be dose dependent at concentrations of beta/A4 peptide ranging from 10 mug/ml to 50 mug/ml. These results suggest that complement components found previously in amyloid deposits may be partly derived from reactive microglia preferentially associated with senile plaques in AD brain.