Wound healing activity of 3-epimasticadienolic acid from Pistachio hulls

Background: Wound healing is a dynamic procedure and normal biological response to the injury which happens through different overlapping phases including inflammation, proliferation and extracellular matrix remodeling phase (maturation). Many inflammatory markers such as, pro-inflammatory cytokines, growth factors, extracellular matrix, and various type cells regulate wound healing process. Wound closure takes place in re-epithelialization phase through keratinocyte and fibroblasts proliferation and migration. For centuries, herbal medicines play an important role in prevention and treatment of wounds and many herbal constituents have played a significant role in discovering novel drug leads. Pistacia vera L. (Pistachio) hulls have been traditionally used as ingredient for oral wound healing and oral antiseptic in Iranian folk medicine, however, its chemical composition with potential wound healing effects is not yet well elucidated.Objective: Aim of this study was to investigate the wound healing activity of one of the major compounds that isolated from potent sub-fraction of Pistachio hulls.Methods: For this purpose we have used in vitro model scratching assay and CFSC dilution assay, respectively, for quantification of fibroblast (NIH3T3 cells) migration and proliferation. The gene expression of two main inflammatory markers including IL-6 and TNf-α was evaluated using Real Time PCR. Isolation and purification of the active compound was performed by column and plate chromatography and its chemical structure was characterized by different methods including 1H, 13C- NMR and mass spectroscopy.Results: The results of this study confirmed that Pistacia vera L. (Pistachio) hulls extract includes bioactive phytosterols such as 3-epimasticadienolic acid that can increase fibroblast migration and proliferation and reduce the scratch area (in vitro model) about 45% at the concentration 200 μg/ml. In addition, this compound at the same dose had significant inhibitory effects on gene expression of IL-6 (P<0.05) and TNF-α (P<0.001) in comparison with untreated cells.Conclusions: Our results supported the ethnopharmacological use of Pistachio hulls in Iranian folk medicine and confirmed the potential wound healing effect of Pistachio hull can be due to the presence of some active phytosterols such as 3-epimasticadienolic acid.

Keywords:

Pistachio hull , wound healing , fibroblast , inflammatory markers , 3-epimasticadienolic acid