DOI: 10.14457/tu.the.2017.1323
title: Antioxidant and anti-inflammatory activities of Thai traditional remedy called Ya-Hom Kae-Lom-Wing-Wien and its ingredients

description: Ya-Hom KAE-LOM-WING-WIEN remedy (KLWW) has been used for treatment of dizziness, fatigue, and sleepiness. It is registered to be a remedy on the Thai National List of Essential Medicines. A.D. 2013. Ya-Hom is nice-smelling herbal remedy and consists of various proportions of twenty-three medicinal ingredients. There are Glycyrrhiza glabra L. (root), Myristica fragrans Houtt. (heartwood), Syzygium aromaticum Merr. et Perry (flower-bud), Angelica sinensis (Oliv.) Diels (root), Ligusticum sinense Oliv. cv.Chuanxiong Hort (rhizome), Vetiveria zizanioides (L.) Nash ex Small (root), Nelumbo nucifera Gaertn. (pollen), Cinnamomum bejolghota (Buch.-Ham.) Sweet (bark), Cinnamomum loureirii Nees. (bark), Cinnamomum  verum J. Presl. (bark), Aquilaria crassna Pierre ex Lecomte. (wood), Euphorbia antiquorum L. (heartwood), Artemisia annua L. (aerial part), Terminalia chebula Retz. var chebula (gall), Alyxia reinwardtii Blume var. lucida Markgr. (bark), borneo camphor, Mimusops elengi L. (wood), Mesua ferrea Linn. (flower), Mimusops elengi L. (flower), Mammea siamensis Kosterm (flower), Ecdysanthera  rosea Hook. & Arn. (vine), sodium borate ,and Dracaena loureiri Gagnep. (heartwood). The aim of this study was to investigate the antioxidant and anti-inflammatory activities of KLWW and its ingredients against super oxide anion, malondialdehyde, and nitric oxide release for preventing and treating chronic diseases.The KLWW consists of four extractions. The ethanolic extracts were extracted by maceration in 95% and 70% ethanol. The aqueous extract was extracted by decoction. The hydrolyzed extract was obtained from aqueous extract, which was extracted similar to physiology of gastro-intestinal tract. The results of percentage of yield the 70% ethanolic extracts had the highest percentage of yield (%yield = 20.86). The ingredients were extracted by maceration in 95% ethanol.The raw materials of plant ingredients were controlled quality following Thai Herbal Pharmacopoeia (THP) standard. All of plant ingredients were investigated quality control methods except borneo camphor and sodium borate. However, KLWW following THP criteria showed loss on drying value of 6.40±0.20%, total ash value of 6.77±0.48%, and acid insoluble ash value of 0.91±0.11%.For antioxidant activity, the extracts of KLWW and its ingredients were investigated by nitroblue tetrazolium (NBT) dye reduction assay in HL-60 cell lines. The 95% ethanolic extract of Mammea siamensis Kosterm showed the highest inhibition by IC50 value of 13.51±2.20 µg/ml, followed by the 95% ethanolic extract of Ecdysanthera rosea. Hook. & Arn. and Terminalia chebula Retz. var chebula with IC50 values of 25.68±2.26 and 32.02±2.36 µg/ml, respectively.Antioxidant activity, the extracts of KLWW and its ingredients were also investigated by thiobarbituric acid reactive substances (TBARS) assay. The hydrolyzed extract of KLWW showed stronger inhibition of TBARS formation by MDA value of 7.72±0.91 nM than propyl gallate (positive control) with MDA value of 10.42±0.66 nM. The 95% ethanolic extract of Mimusops elengi L. showed the highest inhibition of TBARS formation of ingredients with MDA value of (-1.64)±3.22 nM.Anti-inflammatory activity, the extracts of KLWW and its ingredients were investigated by nitric oxide (NO) inhibitory assay in RAW 264.7 cell lines. The 95% extract of KLWW exhibited NO production with IC50 value of 19.53±1.95 µg/ml and the highest inhibition NO of ingredients was the 95% ethanolic extract of Mammea siamensis Kosterm showing IC50 value of 17.13±0.84 µg/ml.Gas chromatography-mass spectrometry (GC-MS) (Thermo Focus GC + polaris Q + auto injector) with column thermo TG-5silms, KLWW of the hydrolyzed, 70% ethanolic, and aqueous extracts showed eugenol as a main compound (42.02, 34.89, and 28.80 % area, respectively) and the 95% ethanolic extract showed that borneol as a main compound (33.31 % area). These results are new knowledge about specific fingerprint profiles and quantification of characteristic compounds in KLWW.For stability study, the 95% ethanolic extract of KLWW was tested by anti-inflammatory activity using NO inhibitory assay. The 95% ethanolic extract of KLWW was stable at least two months because there was significant difference between control and the IC50 values of extract with p-value &lt; 0.05. It lost anti-inflammatory activity within two months but after that it was stable until day 180. However, it has less anti-inflammatory activity than day 0 one time.In conclusion, the hydrolyzed extract of KAE-LOM-WING-WIEN remedy had antioxidant activity by TBARS assay, and the 95% extract of KLWW had anti-inflammatory activity by NO inhibitory assay. All results supported the use of KLWW for reducing free radicals and inflammation. Moreover, KLWW has a strong protective role against oxidative stress and reducing risk of progress in chronic diseases. Therefore, its ethanolic and aqueous extracts should be continuously studied in vitro and in vivo biological activities and using eugenol and borneol as markers to develop further.