The Wound Healing Property of N-Methyl-(2S,4R)-trans-4-Hydroxy-L-Proline from Sideroxylon obtusifolium is Related to its Anti-Inflammatory and Antioxidant Actions

Abstract

Wound healing involves the interaction of blood cells, proteins, proteases, growth factors, and extracellular matrix components. Inflammation is one of the first events occurring during this process. Previously, we showed that the N-Methyl-(2S,4R)-trans-4-Hydroxy-L-Proline (NMP) from Sideroxylon obtusifolium leaves (a Brazilian medicinal species) presents an anti-inflammatory action. Considering inflammation as an important event in the wound healing process, the objectives were to investigate the topical effects of the NMP gel on a mice wound-induced model. Male Swiss mice were divided into 4 groups: Sham (surgical procedure only), Control (gel-base treated), and 3% or 10% NMP gel-treated groups. Measurements of wound areas and microscopic analyses (HE [hematoxylin-eosin] and PSR [picrosirius red] stainings) were carried out, at the 7th and 12th, days after the wound induction. Furthermore, immunohistochemical assays for iNOS (inducible nitric oxide synthase) and COX-2 (cyclooxygenase-2) and biochemical measurements for TBARS (thiobarbituric acid reactive substances), GSH (glutathione), and myeloperoxidase (MPO) were also performed, at the second day after the wound induction. The work showed that NMP decreases the wound areas, after topical application, relatively to the Sham and Control groups. In addition, microscopic alterations were reduced and collagen deposition was increased, at the 7th and 12th days, in the 10% NMP group. While iNOS and COX-2 immunostainings and GSH contents increased, in relation to the Sham and Control groups, TBARS and MPO decreased. Altogether, the results showed NMP to improve the wound healing process, by upregulating iNOS and COX-2 activities, reducing lipid peroxidation and MPO activity, and increasing GSH contents. In addition, NMP certainly contributes to the increased collagen deposition. These data may stimulate translational studies dealing with the possible use of NMP from Sideroxylon obtusifolium or from other sources for the management of wound healing.

Iridoids and Amino Acid Derivatives from the Paraguayan Crude Drug Adenocalymma marginatum (ysypó hû)

“…The occurrence of compound I was confirmed by the MS analysis, in full agreement with the literature [17,18,35]. The compound has been previously reported in Sideroxylon obtusifolium (Sapoteaceae), a medicinal species from Northeast Brazil with anti-inflammatory [36] and wound healing properties [37]. The authors reported that the compound at doses ranging from 25-100 mg/kg decreased licking time and paw edema in the formalin test, as well as a complete reversion of the increased number of polymorphonuclear cells in the inflammation point.…”

Human keratinocyte (HaCaT) stimulation and healing effect of the methanol fraction from the decoction from leaf fromSideroxylon obtusifolium(Roem. & Schult.) T.D. Penn on experimental burn wound model

“…Similarly, a phytomodulatory hydrogel with hemicelluloses extracted from seeds of Caesalpinia pulcherrima (Fabaceae) and mixed with phytomodulatory proteins obtained from the latex of Calotropis procera, enhanced the healing effects by high collagen deposition and modulating some aspects of the inflammatory phase (Vasconcelos et al, 2018). Corroborating with this data, it was discovered that the methanol fraction of quixaba leaves (MFSOL), contains as a main component, the NMP, a derivative from L-proline (amino acid precursor of collagen), that showed cicatrizant effect in the topical treatment of induced excisional lesions in mice, through modulation of the inflammatory response, and increase of the antioxidant response and stimulation of the production of collagen fibers (Aquino et al, 2019). Also, a study with the ethanol extract of the leaves of Passiflora edulis reproduced an acceleration the repair, to reduce the number of inflammatory cells on the 7th day of treatment and to increase the number of fibroblasts and the deposition and organization of the fibers of collagen in 14 days of repair of burns in mice (Barros, Santos, Coelho, Reis, & Bezerra, 2016).…”

“…Other study demonstrated antinociceptive and anti-inflammatory properties of L-proline, N-methyl-(2S, 4R)-trans-4-hydroxy-L-proline (NMP), presents in major proportion in the methanol fraction of leaves of S. obtusifolium (MFSOL) (Aquino et al, 2017). Also, NMP showed wound healing activity on excisional wounds, which was related to its anti-inflammatory and antioxidant actions (Aquino et al, 2019). However, there is no knowledge of whether methanol fraction MFSOL is the healing process in wounds caused by burns.…”

The N-Methyl-(2S, 4R)-trans-4-hydroxy-L-proline-Enriched Methanol Fraction from Sideroxylon obtusifolium Shows an Anticonvulsant Activity Associated with its Anti-inflammatory/Antioxidant Actions

“…Penn. belongs to the Sapotaceae family and is a medicinal plant used in Brazil due to its analgesic and anti-inflammatory activities, as already demonstrated by us [7,8] and others [9]. Furthermore, experimental and clinical evidence indicates that there is a close relationship between epileptogenesis and brain inflammation.…”

“…a DG: (a) vs. control, p < 0.001; (b) vs. Pilo + NMP100, p < 0.001; (c) vs.Pilo + NMP200, p < 0.01; (d) vs. control, p < 0.05. b tC: (a) vs. control, p < 0.0001; (b) vs. Pilo + NMP100, p < 0.0001; (c) vs. Pilo + NMP200, p < 0.0001. c CA1: (a) vs. control, p < 0.001; (b) vs. Pilo + NMP100, p < 0.001; (c) vs. Pilo + NMP200, p < 0.001; (d) vs. control, p < 0.05. d CA3: (a) vs. control, p < 0.001; (b) vs. Pilo + NMP100, p < 0.001; (c) vs. Pilo + NMP200, p < 0.001. (one-way ANOVA and Tukey multiple comparison test).Plant material and preparation of the methanol fraction containing the bioactive constituent N-methyl-(2S,4R)-trans-4-hydroxy-L-proline S. obtusifolium leaves were collected and handled as previously described[8] and identified (voucher specimen #10,648) by Maria Arlene Pessoa da Silva, Ph.D., botanist at the Herbarium "Dárdano de Andrade Lima", Regional University of Cariri (URCA), Crato, Brazil.…”