Anti inflammatory and anti oxidant properties of purified stilbene and crude extracts of Perlis vitis vinifera on cultured huvecs

Abstract

Air-dried and particle form of stems and leaves of Vitis vinifera cultivated in Perlis, Malaysia were extracted using cold, back, methanolic and ethanolic extraction methods. The ethyl acetate stems extract by back extraction (STBE EA) possessed the highest total phenolic content (916.27 ± 0.01) by Folin-Ciocalteu assay and radical scavenging activity at IC50 = 18 µg/mL by DPPH assay. The extracts was further investigated by thin layer chromatography resulted in detection of resveratrol at Rf = 0.425 in STBE EA using petroleum ether:ethyl acetate at ratio 2:3. Advanced screening method of phenolic profiling by LC-MS/MS has detected seven phenolic compounds which were identified through LC-MS/MS library and Mass Bank Software. The compounds are trans-resveratrol, trans-cinnamic acid, e-viniferin, salicylic and caffiec acid, catechin, and epigallocatechin. Purification of the STBE EA has resulted in the isolation of trans-stilbene which was identified as 3, 4’, 5- trihydroxystilbene (trans-resveratrol) by Nuclear Magnetic Resonance. The STBE EA and trans-resveratrol exhibited protective effects on HUVECs against oxidative cell damage by hydrogen peroxide (IC50 = 205 µM) with survival rate 17.69% (IC50 = 1 µg/mL) and 40.80% (IC50 = 0.01 µM) and induced nitric oxide 3.41 (IC50 = 0.001 µg/mL) and 3.96 mm (IC50 = 0.001 µM) respectively. STBE EA and trans-resveratrol possessed dual inhibition of COX-2 and 15-LOX 85.07 ± 1.20 (COX-2) and 60% with IC50 at 7.5 µM (15- LOX) respectively. Furthermore purified trans-resveratrol exhibited better inhibition at 96.87 ± 0.10% for COX-2 and 79.57% for 15-LOX. This study demonstrated that STBE EA extracts and trans-resveratrol isolated from Vitis vinifera stems posses potent anti-oxidant and anti-inflammatory properties.