Sipahi Hande, Orak Duygu, Reis Rengin, Yalman Kübra, Şenol Onur, Palabiyik-Yücelik Saziye Sezin, Deniz İnci, Algül Derya, Guzelmeric Etil, Çelep M Engin, Argin Sanem, Özkan Ferda, Halıcı Zekai, Aydın Ahmet, Yesilada Erdem
Yeditepe University, Faculty of Pharmacy, Department of Toxicology, Istanbul, Turkey.
Yeditepe University, Faculty of Pharmacy, Drug, Cosmetic and Medical Device Research-Development and Analysis Laboratory, Istanbul, Turkey.
J Ethnopharmacol. 2022 Apr 6;287:114843. doi: 10.1016/j.jep.2021.114843. Epub 2021 Nov 18.
Okra fruit (Abelmoschus esculentus (L.) Moench) has been extensively used for the treatment of skin damage and subcutaneous tissue abscess for many years in Turkish folk medicine.
In this study, we aimed to investigate the wound healing potential of okra fruit by in vitro and in vivo experimental models in detail. Furthermore, based on the results of experiments, a wound healing formulation was developed and its activity profile was studied.
For this purpose, the phenolic, flavonoid and proanthocyanidin contents and chemical profile of aqueous and ethanolic extracts prepared from okra fruits cultivated in two different locations of Turkey, i.e. Aegean and Kilis regions, were comparatively determined and the tryptophan levels, which is known to be an influential factor in wound healing, were measured. Antioxidant activity of the okra fruit extracts was determined by DPPH test, ABTS radical scavenger activity, iron-binding capacity, total antioxidant capacity and copper reduction capacity assays. Moreover, antibacterial activity potentials of the aqueous and ethanolic extracts of okra fruits were determined. The protective effect of the extracts against HO-induced oxidative stress and anti-inflammatory activity were assessed in HDF (human dermal fibroblast) cells and in RAW 264.7 murine macrophages, respectively. The biocompatibility of the gel formulations prepared with the best performing extract were evaluated by human Epiderm™ reconstituted skin irritation test model. Wound-healing activity was investigated in rats by in vivo excision model and, histopathological examination of tissues and gene expression levels of inflammation markers were also determined.
According to our findings, the aqueous and ethanolic extracts of okra fruits were found to possess a rich in phenolic content. Besides, isoquercitrin was found to be a marker component in ethanolic extracts of okra fruits. Both extracts exhibited antioxidant activity with significant protective effect against HO-induced damage in HDF cells by diminishing the MDA level. Also, the highest dose of ethanolic extracts has displayed a potent anti-inflammatory activity on LPS-induced RAW264.7 cells. Besides, both water and ethanolic extracts were shown to possess antimicrobial activity. On the other hand, the formulations prepared from the extracts were found non-irritant on in vitro Epiderm™-SIT. In vivo excision assay showed that tissue TGF-β and IL-1β levels were significantly decreased by the 5% okra ethanolic gel formulation. The histopathological analysis also demonstrated that collagenisation and granulation tissue maturation were found higher in 5% (w/v) okra ethanolic extract-treated group.
5% of okra ethanolic extract might be suggested as a potent wound healing agent based on the antimicrobial, antioxidant and anti-inflammatory tests. The proposed activity was also confirmed by the histopathological findings and gene expression analysis.
多年来,秋葵果实(黄葵)在土耳其民间医学中被广泛用于治疗皮肤损伤和皮下组织脓肿。
在本研究中,我们旨在通过体外和体内实验模型详细研究秋葵果实的伤口愈合潜力。此外,根据实验结果,开发了一种伤口愈合制剂并研究了其活性特征。
为此,比较测定了从土耳其两个不同地区即爱琴海地区和基利斯地区种植的秋葵果实制备的水提取物和乙醇提取物中的酚类、黄酮类和原花青素含量及化学特征,并测量了已知对伤口愈合有影响的色氨酸水平。通过DPPH试验、ABTS自由基清除活性、铁结合能力、总抗氧化能力和铜还原能力测定法测定秋葵果实提取物的抗氧化活性。此外,还测定了秋葵果实水提取物和乙醇提取物的抗菌活性潜力。分别在人皮肤成纤维细胞(HDF)和RAW 264.7小鼠巨噬细胞中评估提取物对过氧化氢(HO)诱导的氧化应激的保护作用和抗炎活性。用性能最佳的提取物制备的凝胶制剂的生物相容性通过人表皮™重组皮肤刺激试验模型进行评估。通过体内切除模型在大鼠中研究伤口愈合活性,并确定组织的组织病理学检查和炎症标志物的基因表达水平。
根据我们的研究结果,发现秋葵果实的水提取物和乙醇提取物富含酚类物质。此外,异槲皮苷被发现是秋葵果实乙醇提取物中的一种标志性成分。两种提取物均表现出抗氧化活性,通过降低丙二醛(MDA)水平对HDF细胞中HO诱导的损伤具有显著的保护作用。此外,最高剂量的乙醇提取物对脂多糖(LPS)诱导的RAW264.7细胞显示出强大的抗炎活性。此外,水提取物和乙醇提取物均显示具有抗菌活性。另一方面,从提取物制备的制剂在体外表皮™-SIT试验中显示无刺激性。体内切除试验表明,5%秋葵乙醇凝胶制剂可显著降低组织转化生长因子-β(TGF-β)和白细胞介素-1β(IL-1β)水平。组织病理学分析还表明,5%(w/v)秋葵乙醇提取物处理组的胶原化和肉芽组织成熟度更高。
基于抗菌、抗氧化和抗炎试验,5%的秋葵乙醇提取物可能被认为是一种有效的伤口愈合剂。组织病理学结果和基因表达分析也证实了所提出的活性。