Anwar Mohamed A, El Gedaily Rania A, Salama Abeer, Aboulthana Wael M, Kandil Zeinab A, Abdel-Dayem Shymaa I A
Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Kasr El Aini St., P.B. 11562, Cairo, Egypt.
Pharmacology Department, Medical Research and Clinical Studies Institute, National Research Centre, 33 El Bohouth St., P.O. 12622, Dokki, Giza, Egypt.
J Ethnopharmacol. 2025 Jan 30;337(Pt 3):118983. doi: 10.1016/j.jep.2024.118983. Epub 2024 Oct 28.
Scientific publications documented the use of plants from Genus Malva to treat inflammatory diseases and skin disorders by our ancestors. Malva parviflora L. has reported benefits for wound healing in traditional medicine; however, there is a lack of experimental study to validate these claims.
We initiated this study to explore the metabolites and verify the wound healing properties of M. parviflora using in vivo and in vitro models.
Liquid chromatography electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS) was used to identify the ethanolic extract different metabolites. Additionally, total phenolic content was determined using Folin-Ciocalteu reagent. To verify the extract wound healing potential, an in vivo rat wound excision model was employed. Round wounds (5 mm in diameter) were created by a sterile biopsy punch needle. The wounds were treated with plant extracts (2.5% and 5%) as well as a commercially available wound healing product (Mebo®) for 10 days. The results were assessed as follows: 1) Measuring the reduction% in wound area compared to the original wound size. 2) Evaluation of the levels of wound healing biomarkers, namely collagen type I (Col-1), alpha smooth muscle actin (α-SMA), extracellular signal-regulated kinases-1 (ERK1), and matrix metalloproteinase-9 (MMP9) levels. 3) Performing histopathological examination of the wound tissue. The antioxidant properties of the M. parviflora leaves ethanolic extract were investigated using various assays: total antioxidant capacity (TAC), iron reducing power (IRP), 1,1-Diphenyl-2-picryl-hydrazyl (DPPH), and 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radicals scavenging assays. Furthermore, the anti-inflammatory activity was confirmed by calculating the inhibition percentages of protein denaturation and the activity of the proteinase enzyme.
Liquid chromatography tandem mass spectrometry (LC-MS/MS) analysis revealed the presence of various secondary metabolites in M. parviflora ethanolic extract, including phenolic acids (cinnamic and ferulic acids), flavonoids (quercetin and "iso"rhamnetin monoglucuronides), fatty acids (hydroxy-octadecatrienoic and oxo-octadecatrienoic acids), in addition to chlorophyll derivatives and carotenoids (pheophorbide-a and lutein, respectively). Malva extracts significantly reduced wound size compared to untreated control group. The extracts also promoted wound healing by upregulating collagen I, α-SMA, and ERK1 levels, while downregulating MMP9 expression. Notably, the effect of 2.5% and 5% extracts was similar or exceeds those of Mebo®, supported by histopathological results. Finally, M. parviflora ethanolic extract exhibited antioxidant and anti-inflammatory potentials comparable to the used standards.
Our study provides evidence-based support for the wound healing properties of M. parviflora L. leaves ethanolic extract. This is further strengthened by the fact that many of the identified metabolites possess wound healing, antioxidant, and/or anti-inflammatory activities.
科学出版物记载了我们的祖先使用锦葵属植物治疗炎症性疾病和皮肤疾病。小花锦葵在传统医学中据报道对伤口愈合有益；然而，缺乏实验研究来证实这些说法。
我们开展这项研究，以探索小花锦葵的代谢产物，并使用体内和体外模型验证其伤口愈合特性。
采用液相色谱电喷雾电离串联质谱法（LC/ESI-MS/MS）鉴定乙醇提取物中的不同代谢产物。此外，使用福林-酚试剂测定总酚含量。为验证提取物的伤口愈合潜力，采用体内大鼠伤口切除模型。用无菌活检穿刺针制作直径5毫米的圆形伤口。伤口分别用植物提取物（2.5%和5%）以及一种市售伤口愈合产品（美宝®）处理10天。结果评估如下：1）测量伤口面积相对于原始伤口大小的减少百分比。2）评估伤口愈合生物标志物的水平，即I型胶原蛋白（Col-1）、α平滑肌肌动蛋白（α-SMA）、细胞外信号调节激酶-1（ERK1）和基质金属蛋白酶-9（MMP9）的水平。3）对伤口组织进行组织病理学检查。采用多种测定方法研究小花锦葵叶乙醇提取物的抗氧化性能：总抗氧化能力（TAC）、铁还原能力（IRP）、1,1-二苯基-2-苦基肼（DPPH）和2,2'-偶氮双（3-乙基苯并噻唑啉-6-磺酸）（ABTS）自由基清除测定。此外，通过计算蛋白质变性的抑制百分比和蛋白酶的活性来确认抗炎活性。
液相色谱串联质谱（LC-MS/MS）分析表明，小花锦葵乙醇提取物中存在多种次生代谢产物，包括酚酸（肉桂酸和阿魏酸）、黄酮类化合物（槲皮素和“异”鼠李素单葡萄糖醛酸苷）、脂肪酸（羟基十八碳三烯酸和氧代十八碳三烯酸），此外还有叶绿素衍生物和类胡萝卜素（分别为脱镁叶绿酸-a和叶黄素）。与未处理的对照组相比，锦葵提取物显著减小了伤口大小。提取物还通过上调I型胶原蛋白、α-SMA和ERK1水平，同时下调MMP9表达来促进伤口愈合。值得注意地是，2.5%和5%提取物的效果与美宝®相似或超过美宝®，组织病理学结果支持这一点。最后，小花锦葵乙醇提取物表现出与所用标准相当的抗氧化和抗炎潜力。
我们的研究为小花锦葵叶乙醇提取物的伤口愈合特性提供了循证支持。许多已鉴定的代谢产物具有伤口愈合、抗氧化和/或抗炎活性这一事实进一步强化了这一点。
