Anti-oxidative assays as markers for anti-inflammatory activity of flavonoids.
The complexity of in vitro anti-inflammatory assays, the cost and time consumed, and the necessary skills can be a hurdle to apply to promising compounds in a high throughput setting. In this study, several antioxidative assays i.e. DPPH, ABTS, ORAC and xanthine oxidase (XO) were used to examine the antioxidative activity of three sub groups of flavonoids: (i) flavonol: quercetin, myricetin, (ii) flavanone: eriodictyol, naringenin (iii) flavone: luteolin, apigenin. A range of flavonoid concentrations was tested for their antioxidative activities and were found to be dose-dependent. However, the flavonoid concentrations over 50ppm were found to be toxic to the THP-1 monocytes. Therefore, 10, 20 and 50ppm of flavonoid concentrations were tested for their anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated THP-1 monocytes. Expression of inflammatory genes, IL-1β, IL-6, IL-8, IL-10 and TNF-α was found to be sequentially decreased when flavonoid concentration increased. Principle component analysis (PCA) was used to investigate the relationship between the data sets of antioxidative assays and the expression of inflammatory genes. The results showed that DPPH, ABTS and ORAC assays have an opposite correlation with the reduction of inflammatory genes. Pearson correlation exhibited a relationship between the ABTS assay and the expression of three out of five analyzed genes; IL-1β, IL-6 and IL-8. Our findings indicate that ABTS assay can potentially be an assay marker for anti-inflammatory activity of flavonoids.
Department of Food Science and Technology, Faculty of Agro-Industry, Kasetsart University, 50 Ngamwongwan Rd., Ladyoaw, Chatuchuck, Bangkok 10900, Thailand. Electronic address: wasaporn.c@ku.ac.th.
Wasaporn
W
Chanput
Department of Food Science and Technology, Faculty of Agro-Industry, Kasetsart University, 50 Ngamwongwan Rd., Ladyoaw, Chatuchuck, Bangkok 10900, Thailand.
Narumol
N
Krueyos
Department of Product Development, Faculty of Agro-Industry, Kasetsart University, 50 Ngamwongwan Rd., Ladyoaw, Chatuchuck, Bangkok 10900, Thailand.
Pitiporn
P
Ritthiruangdej
D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology; D052160:Benzothiazoles / Q000737:chemistry; D001713:Biphenyl Compounds / Q000737:chemistry; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000235:genetics; D005419:Flavonoids / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D008070:Lipopolysaccharides; D010851:Picrates / Q000737:chemistry; D013451:Sulfonic Acids / Q000737:chemistry; D014969:Xanthine Oxidase / Q000737:chemistry
D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D052160:Benzothiazoles; D001713:Biphenyl Compounds; D016207:Cytokines; D005419:Flavonoids; D008070:Lipopolysaccharides; D010851:Picrates; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; C004931:1,1-diphenyl-2-picrylhydrazyl; D014969:Xanthine Oxidase
Anti-inflammation; Anti-oxidative assay; Correlation; Flavonoid