Identification of Eupatilin from Artemisia argyi as a Selective PPARα Agonist Using Affinity Selection Ultrafiltration LC-MS

Abstract

Peroxisome proliferator-activated receptors (PPARs) are key nuclear receptors and therapeutic targets for the treatment of metabolic diseases through the regulation of insulin resistance, diabetes, and dyslipidemia. Although a few drugs that target PPARs have been approved, more diverse and novel PPAR ligands are necessary to improve the safety and efficacy of available drugs. To expedite the search for new natural agonists of PPARs, we developed a screening assay based on ultrafiltration liquid chromatography-mass spectrometry (LC-MS) that is compatible with complex samples such as dietary foods or botanical extracts. The known PPARα and/or PPARγ ligands resveratrol and rosiglitazone were used as positive controls to validate the developed method. When applied to the screening of an Artemisia argyi extract, eupatilin was identified as a selective PPARα ligand. A PPAR competitive binding assay based on FRET detection also confirmed eupatilin as a selective PPARα agonist exhibiting a binding affinity of 1.18 μM (IC50). Furthermore, eupatilin activation of the transcriptional activity of PPARα was confirmed using a cell-based transactivation assay. Thus, ultrafiltration LC-MS is a suitable assay for the identification of PPAR ligands in complex matrixes such as extracts of dietary foods and botanicals.

Protective Effect of Phenolic Compounds Isolated from Mugwort (Artemisia argyi) against Contrast-Induced Apoptosis in Kidney Epithelium Cell Line LLC-PK1

“…Artemisia argyi , an aromatic herb belonging to the Asteraceae family, has been mainly used in traditional oriental medicine for treating various diseases, such as menstrual disorders, infertility, uterine bleeding, and inflammatory diseases [13,14,15,16,17]. In addition, the extract of A. argyi is known to reduce ethanol-induced gastrointestinal damage [18], carbon tetrachloride (CCl 4 )-induced hepatic damage [19], and cerulein-induced pancreatic damage [20] in rats.…”

Rapid screening and characterization of acetylcholinesterase inhibitors from Yinhuang oral liquid using ultrafiltration-liquid chromatography-electrospray ionization tandem mass spectrometry

“…According to the methods in previous studies,[ 14 15 16 ] 200 μL sample solution was added into a 24-well plate together with 100 μL acetylcholinesterase enzyme solution with the concentration of 10U/mL. After adding 200 μL PBS buffer, sample was incubated at 37°C for 30 min.…”

Exploration on the Interaction Ability of Antitumor Compound Bis-[2,6-difluoro-N-(hydroxyl-<κ>O)benzamidato-<κ>O]dibutylitin(IV) with Human Peroxisome Proliferator-Activated Receptor hPPARγ

“…Based on the ultraviolet absorption of the compound, the DBDF2,6T which were bound to hPPAR γ could be detected through analyzing the washed solution by liquid chromatography. This method was first used in such confirmatory experiment and marked by its simplicity, generality, and applicability [ 18 ].…”