Inhibition of beta-amyloid(1-40) Peptide Aggregation and Neurotoxicity by Citrate.
The accumulation of beta-amyloid (Abeta) aggregates is a characteristic of Alzheimer's disease (AD). Furthermore, these aggregates have neurotoxic effects on cells, and thus, molecules that inhibit Abeta aggregate formation could be valuable therapeutics for AD. It is well known that aggregation of Abeta depends on its hydrophobicity, and thus, in order to increase the hydrophilicity of Abeta, we considered using citrate, an anionic surfactant with three carboxylic acid groups. We hypothesized that citrate could reduce hydrophobicity and increase hydrophilicity of Abeta(1-40) molecules via hydrophilic/electrostatic interactions. We found that citrate significantly inhibited Abeta(1-40) aggregation and significantly protected SH-SY5Y cell line against Abeta(1-40) aggregates-induced neurotoxicity. In details, we examined the effects of citrate on Abeta(1-40) aggregation and on Abeta(1-40) aggregates-induced cytotoxicity, cell viability, and apoptosis. Th-T assays showed that citrate significantly inhibited Abeta(1-40) aggregation in a concentration-dependent manner (Th-T intensity: from 91.3% in 0.01 mM citrate to 82.1% in 1.0 mM citrate vs. 100.0% in Abeta(1-40) alone). In cytotoxicity and viability assays, citrate reduced the toxicity of Abeta(1-40) in a concentration-dependent manner, in which the cytotoxicity decreased from 107.5 to 102.3% as compared with Abeta(1-40) aggregates alone treated cells (127.3%) and the cell viability increased from 84.6 to 93.8% as compared with the Abeta(1-40) aggregates alone treated cells (65.3%). Furthermore, Hoechst 33342 staining showed that citrate (1.0 mM) suppressed Abeta(1-40) aggregates-induced apoptosis in the cells. This study suggests that citrate can inhibit Abeta(1-40) aggregation and protect neurons from the apoptotic effects of Abeta(1-40) aggregates. Accordingly, our findings suggest that citrate administration should be viewed as a novel neuroprotective strategy for AD.
Park
Yong Hoon
YH
Inam Neuroscience Research Center, Sanbon Medical Center, College of Medicine, Wonkwang University, Gunpo 435-040, Korea.
Kim
Young-Jin
YJ
Son
Il Hong
IH
Yang
Hyun Duk
HD