3,3',4',5'-Tetrahydroxyflavone induces formation of large aggregates of amyloid β protein.
Amyloid β protein (Aβ) self-assembles into insoluble fibrils, and forms the senile plaques associated with Alzheimer's disease. 3,3',4',5'-Tetrahydroxyflavone, a synthetic analogue of the natural flavonoid fisetin, has been found to potently inhibit Aβ fibril formation. In the present study, we investigated how inhibition of Aβ fibril formation by this flavonoid affects Aβ conformation and neurotoxicity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of Aβ1-42 (20 µM) incubated with or without 3,3',4',5'-tetrahydroxyflavone demonstrated that 3,3',4',5'-tetrahydroxyflavone (100 µM) rapidly caused formation of atypical Aβ conformers, which appeared as a very broad, smear-like band in the high molecular weight region and were distinguishable from soluble Aβ oligomers or mature Aβ fibrils. Transmission electron microscopy (TEM) revealed that large spherical Aβ aggregates were preferentially formed in the presence of 3,3',4',5'-tetrahydroxyflavone. The SDS-resistant, smear-like band on SDS-PAGE and the large spherical aggregates in TEM both disappeared after heat treatment (100°C, 10 min). Furthermore, a neurotoxicity assay with cultured rat hippocampal neurons demonstrated that Aβ incubated with 3,3',4',5'-tetrahydroxyflavone was significantly less toxic than Aβ incubated without the flavonoid. These results suggest that the newly synthesized fisetin analogue 3,3',4',5'-tetrahydroxyflavone directly produces atypical, large Aβ aggregates and reduces Aβ toxicity.
Laboratory of Pharmacology, Faculty of Pharmacy and Research Institute of Pharmaceutical Sciences, Musashino University.
Hiroko
H
Ushikubo
Yui
Y
Tanimoto
Kazuho
K
Abe
Tomohiro
T
Asakawa
Toshiyuki
T
Kan
Tatsuhiro
T
Akaishi
D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D047309:Flavones / Q000494:pharmacology*; D005419:Flavonoids / Q000494:pharmacology; D044948:Flavonols; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D061251:Primary Cell Culture; D066329:Protein Aggregates / Q000187:drug effects*; D011487:Protein Conformation / Q000187:drug effects; D051381:Rats
C000595791:3,3',4',5'-tetrahydroxyflavone; D016229:Amyloid beta-Peptides; D047309:Flavones; D005419:Flavonoids; D044948:Flavonols; D066329:Protein Aggregates; C017875:fisetin