Beijing team builds AD12 cell line and screens 16 alkaloids; aconite extract eased symptoms in 5xFAD mice

A team from the Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences (Beijing), led by Haiyu Zhao and Xiaolu Wei, has described a stepwise strategy for finding anti-Alzheimer's compounds. The work was published in Science of Traditional Chinese Medicine; the authors report a new cell line, a screen of 16 alkaloids and a test in transgenic mice doi.orgScience of Traditional Chinese Medicinedoi.orgOpen source ↗. All results are preclinical: cell cultures and mice.

Cell model and alkaloid screen

The authors note that existing cell models for drug screening mostly rely on single-gene transfection and do not reproduce the complex pathology of the disease. Transgenic animals mimic it better but are slow, costly and variable between individuals.

The team used lentiviral transfection to create a U251MG cell line stably expressing the APP695/PS1-ΔE9 double transgene and named it AD12. The cells were stimulated with lipopolysaccharide (LPS), which induced neuroinflammation: levels of interleukin-6, amyloid-β 1-42 (Aβ 1-42) and PS1 rose. Metabolomic analysis showed that the main affected pathways were alanine, aspartate and glutamate metabolism and arginine biosynthesis.

Sixteen alkaloids derived from traditional Chinese medicine were screened in the LPS-AD12 model. Benzoylaconitine (BAC) significantly reduced Aβ 1-42 and interleukin-6. Molecular docking indicated strong binding of BAC to APP and PS1 proteins doi.orgScience of Traditional Chinese Medicinedoi.orgOpen source ↗.

Targeted metabolomics and tests in 5xFAD mice

Targeted metabolomics of amino acids and the tricarboxylic acid cycle in BAC-treated cells linked its activity mainly to anti-inflammatory effects, modulation of cholinergic signaling and regulation of energy metabolism, according to the authors.

Aconite extract was then given to 5xFAD transgenic mice. Behavioral tests and immunofluorescence of brain tissue (Aβ, glial fibrillary acidic protein and ionized calcium-binding adapter molecule 1) showed, the authors report, that the extract significantly alleviated Alzheimer's-related symptoms. Serine, acetylcholine, ornithine and gamma-aminobutyric acid showed consistent trends in both brain tissue and serum of treated animals.

The authors conclude that the AD12 model and LPS-based screen are a reliable system and propose a pipeline of cell model construction, evaluation of individual compounds and identification of effective component groups. They state that it may speed up the search for anti-Alzheimer's agents and serve as a framework for developing drugs from traditional Chinese medicine doi.orgScience of Traditional Chinese Medicinedoi.orgOpen source ↗.