Medicinal value of asiaticoside for Alzheimer's disease as assessed using single-molecule-detection fluorescence correlation spectroscopy, laser-scanning microscopy, transmission electron microscopy, and in silico docking

BMC Complement Altern Med. 2015 Apr 14;15:118. doi: 10.1186/s12906-015-0620-9.


Background: Identifying agents that inhibit amyloid beta peptide (Aβ) aggregation is the ultimate goal for slowing Alzheimer's disease (AD) progression. This study investigated whether the glycoside asiaticoside inhibits Aβ1-42 fibrillation in vitro.

Methods: Fluorescence correlation spectroscopy (FCS), evaluating the Brownian diffusion times of moving particles in a small confocal volume at the single-molecule level, was used. If asiaticoside inhibits early Aβ1-42 fibrillation steps, more Aβs would remain free and rapidly diffuse in the confocal volume. In contrast, "weaker or no inhibition" permits a greater number of Aβs to polymerize into oligomers, leading to fibers and gives rise to slow diffusion times in the solution. Trace amounts of 5-carboxytetramethylrhodamine (TAMRA)-labeled Aβ1-42 in the presence of excess unlabeled Aβ1-42 (10 μM) was used as a fluorescent probe. Steady-state and kinetic-Thioflavin T (ThT) fluorospectroscopy, laser-scanning fluorescence microscopy (LSM), and transmission electron microscopy (TEM) were also used to monitor fibrillation. Binding of asiaticoside with Aβ1-42 at the atomic level was computationally examined using the Molegro Virtual Docker and PatchDock.

Results: With 1 h of incubation time for aggregation, FCS data analysis revealed that the diffusion time of TAMRA-Aβ1-42 was 208 ± 4 μs, which decreased to 164 ± 8.0 μs in the presence of asiaticoside, clearly indicating that asiaticoside inhibited the early stages Aβ1-42 of fibrillation, leaving more free Aβs in the solution and permitting their rapid diffusion in the confocal volume. The inhibitory effects were also evidenced by reduced fiber formation as assessed by steady-state and kinetic ThT fluorospectroscopy, LSM, and TEM. Asiaticoside elongated the lag phase of Aβ1-42 fibrillation, indicating the formation of smaller amyloid species were impaired in the presence of asiaticoside. Molecular docking revealed that asiaticoside binds with amyloid intra- and inter-molecular amino acid residues, which are responsible for β-sheet formation and longitudinal extension of fibrils.

Conclusion: Finally, asiaticoside prevents amyloidogenesis that precedes neurodegeneration in patients with Alzheimer's disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / metabolism*
  • Alzheimer Disease / prevention & control
  • Amyloid / metabolism*
  • Amyloid beta-Peptides / metabolism*
  • Centella / chemistry*
  • Fluorescence
  • Humans
  • Microscopy / methods
  • Molecular Docking Simulation / methods
  • Peptide Fragments / metabolism*
  • Phytotherapy*
  • Plant Extracts / pharmacology*
  • Plant Extracts / therapeutic use
  • Rhodamines / metabolism
  • Spectrum Analysis / methods
  • Triterpenes / pharmacology*
  • Triterpenes / therapeutic use


  • 5-carboxytetramethylrhodamine
  • Amyloid
  • Amyloid beta-Peptides
  • Peptide Fragments
  • Plant Extracts
  • Rhodamines
  • Triterpenes
  • amyloid beta-protein (1-42)
  • asiaticoside