Activities of the ethanolic extract of Wisumpayayai remedy on lipopolysaccharide-stimulated mcrophages

Wisumpayayai is one of the household traditional remedy approved by the Ministry of Public Health of Thailand in The National Drug List as anti-flatulence and anti-dyspepsia medication. It is composed of 20 herbal plants including; Coriandrum sativum, Diospyros decandra, Myristica fragrans, Amomum krervanh, Syzygium aromaticum, Angelica dahurica, Atractylodes lancea, Conioselinum univitatum, Angelica sinensis, Artemisia pallens, Cinnamomum verum, Cinnamomum bejolghota, Terminalia arjuna, Terminalia chebula, Acorus calamus, Aristolochia sp., Tinospora crispa, Zingiber kerrii, Melastoma saugnineum, Piper retrofractum. Some of the plants in this remedy have been reported to exert anti-inflammatory activities and a variety of the pharmacological properties. No evidences have been found in the anti-inflammatory effect of Wisumpayayai remedy. This study aimed to investigate the effect of the ethanol extract of Wisumpayayai remedy on lipopolysaccharide-stimulated macrophages and to determine its total anti-oxidant activities by FRAP assay. The results demonstrated that the ethanol extracts of Wisumpayayai remedy significantly inhibited phagocytic activity of the activated macrophages by zymosan-NBT reduction assay in a concentration-dependent manner (12.5-100 µg/ml). It also significantly inhibited nitric oxide (NO) production either by pretreatment and cotreatment of the extract and without cytotoxic effect to the cells. Their IC50 values for NO inhibition were 40.77 and 37.39 µg/ml for pre-treatment and co-treatment respectively. The extract 50 and 100µg/m significantly decreased mRNA expression of cyclooxygenase 2 (COX-2) and inducible nitric oxide synthase (iNOS). These enzymes were responsible for PGE2 and NO production in the activated macrophages. Moreover, the extract 50 and 100 µg/ml were able to inhibit mRNA expression of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α as well as chemokines MCP-1, MIP-1α and IL-8. These enzymes and cytokines are inflammatory mediators produced during inflammatory process. The antioxidant assay showed high antioxidant activity of the extracts at 100 µg/ml. The results obtained from this study indicated that Wisumpayayai ethanol extract were able to inhibit activated macrophages functions. Therefore, this remedy may possess anti-inflammatory activity.