Uvaol Improves the Functioning of Fibroblasts and Endothelial Cells and Accelerates the Healing of Cutaneous Wounds in Mice
Abstract
:1. Introduction
2. Results
2.1. Effect of Uvaol on Cell Viability
2.2. Effects of Uvaol Treatment on Fibroblast and Endothelial Cell Motility
2.3. Effect of Uvaol on ECM Deposition by Fibroblasts
2.4. Uvaol Stimulates Tube-Like Structure Formation In Vitro
2.5. Involvement of the PKA and p38-MAPK Signaling Pathways in Uvaol Induced both Fibroblast and Endothelial Cell Motility
2.6. Effects of Uvaol on the In Vivo Wound Healing Assay
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. Cell Culture
4.3. Cell Viability Assay
4.4. Scratch Wound Healing Assay
4.5. Immunofluorescence Staining
4.6. Tube-Like Structure Formation Assay
4.7. Animals
4.8. Mouse Excisional Wound Model
4.9. Wound Contraction Measurements
4.10. Statistical Analysis
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- Sorg, H.; Tilkorn, D.J.; Hager, S.; Hauser, J.; Mirastschijski, U. Skin wound healing: An update on the current knowledge and concepts. Eur. Surg. Res. 2017, 58, 81–94. [Google Scholar] [CrossRef]
- Ammann, K.R.; DeCook, K.J.; Li, M.; Slepian, M.J. Migration versus proliferation as contributor to in vitro wound healing of vascular endothelial and smooth muscle cells. Exp. Cell Res. 2019, 376, 58–66. [Google Scholar] [CrossRef] [PubMed]
- Kanji, S.; Das, H. Advances of stem cell therapeutics in cutaneous wound healing and regeneration. Mediators Inflamm. 2017, 2017, 5217967. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Li, J.; Wang, J.; Wang, Z.; Xia, Y.; Zhou, M.; Zhong, A.; Sun, J. Experimental models for cutaneous hypertrophic scar research. Wound Repair Regen. 2019, 1, 126–144. [Google Scholar] [CrossRef] [PubMed]
- Boniakowski, A.E.; Kimball, A.S.; Jacobs, B.N.; Kunkel, S.L.; Gallagher, K.A. Macrophage-mediated inflammation in normal and diabetic wound healing. J. Immunol. 2017, 199, 17–24. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Tracy, L.E.; Minasian, R.A.; Caterson, E.J. Extracellular matrix and dermal fibroblast function in the healing wound. Adv. Wound Care (New Rochelle) 2016, 5, 119–136. [Google Scholar] [CrossRef]
- Bainbridge, P. Wound healing and the role of fibroblasts. J. Wound Care 2013, 22, 407–408, 410–412. [Google Scholar]
- DiPietro, L.A. Angiogenesis and wound repair: When enough is enough. J. Leukoc. Biol. 2016, 100, 979–984. [Google Scholar] [CrossRef]
- Petrovska, B.B. Historical review of medicinal plants’ usage. Pharm. Rev. 2012, 6, 1–5. [Google Scholar] [CrossRef] [Green Version]
- Shukla, A.; Rasik, A.M.; Jain, G.K.; Shankar, R.; Kulshrestha, D.K.; Dhawan, B.N. In vitro and in vivo wound healing activity of asiaticoside isolated from Centella asiatica. J. Ethnopharmacol. 1999, 65, 1–11. [Google Scholar] [CrossRef]
- Calixto, J.B. The role of natural products in modern drug discovery. An. Acad. Bras. Cienc. 2019, 91, e20190105. [Google Scholar] [CrossRef] [PubMed]
- Agra, L.C.; Ferro, J.N.; Barbosa, F.T.; Barreto, E. Triterpenes with healing activity: A systematic review. J. Dermatolog. Treat. 2015, 26, 465–470. [Google Scholar] [CrossRef]
- Hill, R.A.; Connolly, J.D. Triterpenoids. Nat. Prod. Rep. 2013, 30, 1028–1065. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Sanchez-Quesada, C.; Lopez-Biedma, A.; Warleta, F.; Campos, M.; Beltran, G.; Gaforio, J.J. Bioactive properties of the main triterpenes found in olives, virgin olive oil, and leaves of Olea europaea. J. Agric. Food Chem. 2013, 61, 12173–12182. [Google Scholar] [CrossRef] [PubMed]
- Allouche, Y.; Warleta, F.; Campos, M.; Sanchez-Quesada, C.; Uceda, M.; Beltran, G.; Gaforio, J.J. Antioxidant, antiproliferative, and pro-apoptotic capacities of pentacyclic triterpenes found in the skin of olives on MCF-7 human breast cancer cells and their effects on DNA damage. J. Agric. Food Chem. 2011, 59, 121–130. [Google Scholar] [CrossRef]
- Agra, L.C.; Lins, M.P.; da Silva Marques, P.; Smaniotto, S.; Bandeira de Melo, C.; Lagente, V.; Barreto, E. Uvaol attenuates pleuritis and eosinophilic inflammation in ovalbumin-induced allergy in mice. Eur. J. Pharmacol. 2016, 780, 232–242. [Google Scholar] [CrossRef]
- Luna-Vazquez, F.J.; Ibarra-Alvarado, C.; Rojas-Molina, A.; Romo-Mancillas, A.; Lopez-Vallejo, F.H.; Solis-Gutierrez, M.; Rojas-Molina, J.I.; Rivero-Cruz, F. Role of nitric oxide and hydrogen sulfide in the vasodilator effect of ursolic acid and uvaol from black cherry prunus serotina fruits. Molecules 2016, 21, 78. [Google Scholar] [CrossRef] [Green Version]
- Martin, R.; Ibeas, E.; Carvalho-Tavares, J.; Hernandez, M.; Ruiz-Gutierrez, V.; Nieto, M.L. Natural triterpenic diols promote apoptosis in astrocytoma cells through ROS-mediated mitochondrial depolarization and JNK activation. PLoS ONE 2009, 4, e5975. [Google Scholar] [CrossRef] [Green Version]
- Okamoto, T.; Akita, N.; Kawamoto, E.; Hayashi, T.; Suzuki, K.; Shimaoka, M. Endothelial connexin32 enhances angiogenesis by positively regulating tube formation and cell migration. Exp. Cell Res. 2014, 321, 133–141. [Google Scholar] [CrossRef]
- Canedo-Dorantes, L.; Canedo-Ayala, M. Skin acute wound healing: A comprehensive review. Int. J. Inflam. 2019, 2019, 3706315. [Google Scholar] [CrossRef]
- Li, J.; Zhang, Y.P.; Kirsner, R.S. Angiogenesis in wound repair: Angiogenic growth factors and the extracellular matrix. Microsc. Res. Tech. 2003, 60, 107–114. [Google Scholar] [CrossRef] [PubMed]
- Lerman, O.Z.; Galiano, R.D.; Armour, M.; Levine, J.P.; Gurtner, G.C. Cellular dysfunction in the diabetic fibroblast: Impairment in migration, vascular endothelial growth factor production, and response to hypoxia. Am. J. Pathol. 2003, 162, 303–312. [Google Scholar] [CrossRef]
- Hu, Y.; Rao, S.S.; Wang, Z.X.; Cao, J.; Tan, Y.J.; Luo, J.; Li, H.M.; Zhang, W.S.; Chen, C.Y.; Xie, H. Exosomes from human umbilical cord blood accelerate cutaneous wound healing through miR-21-3p-mediated promotion of angiogenesis and fibroblast function. Theranostics 2018, 8, 169–184. [Google Scholar] [CrossRef]
- Somova, L.I.; Shode, F.O.; Mipando, M. Cardiotonic and antidysrhythmic effects of oleanolic and ursolic acids, methyl maslinate and uvaol. Phytomedicine 2004, 11, 121–129. [Google Scholar] [CrossRef]
- Botelho, R.M.; Tenorio, L.P.G.; Silva, A.L.M.; Tanabe, E.L.L.; Pires, K.S.N.; Goncalves, C.M.; Santos, J.C.; Marques, A.L.X.; Allard, M.J.; Bergeron, J.D.; et al. Biomechanical and functional properties of trophoblast cells exposed to Group B Streptococcus in vitro and the beneficial effects of uvaol treatment. Biochim. Biophys. Acta Gen. Subj. 2019, 1863, 1417–1428. [Google Scholar] [CrossRef]
- Miura, N.; Matsumoto, Y.; Miyairi, S.; Nishiyama, S.; Naganuma, A. Protective effects of triterpene compounds against the cytotoxicity of cadmium in HepG2 cells. Mol. Pharmacol. 1999, 56, 1324–1328. [Google Scholar] [CrossRef]
- Jonkman, J.E.; Cathcart, J.A.; Xu, F.; Bartolini, M.E.; Amon, J.E.; Stevens, K.M.; Colarusso, P. An introduction to the wound healing assay using live-cell microscopy. Cell. Adh. Migr. 2014, 8, 440–451. [Google Scholar] [CrossRef] [Green Version]
- Bernabe-Garcia, A.; Armero-Barranco, D.; Liarte, S.; Ruzafa-Martinez, M.; Ramos-Morcillo, A.J.; Nicolas, F.J. Oleanolic acid induces migration in Mv1Lu and MDA-MB-231 epithelial cells involving EGF receptor and MAP kinases activation. PLoS ONE 2017, 12, e0172574. [Google Scholar] [CrossRef]
- Pereira Beserra, F.; Xue, M.; Maia, G.L.A.; Leite Rozza, A.; Helena Pellizzon, C.; Jackson, C.J. Lupeol, a pentacyclic triterpene, promotes migration, wound closure, and contractile effect in vitro: Possible involvement of PI3K/Akt and p38/ERK/MAPK pathways. Molecules 2018, 23, 2819. [Google Scholar] [CrossRef] [Green Version]
- Liu, X.J.; Kong, F.Z.; Wang, Y.H.; Zheng, J.H.; Wan, W.D.; Deng, C.L.; Mao, G.Y.; Li, J.; Yang, X.M.; Zhang, Y.L.; et al. Lumican accelerates wound healing by enhancing alpha2beta1 integrin-mediated fibroblast contractility. PLoS ONE 2013, 8, e67124. [Google Scholar]
- Hashim, P. The effect of Centella asiatica, vitamins, glycolic acid and their mixtures preparations in stimulating collagen and fibronectin synthesis in cultured human skin fibroblast. Pak. J. Pharm. Sci. 2014, 27, 233–237. [Google Scholar]
- Kim, W.K.; Song, S.Y.; Oh, W.K.; Kaewsuwan, S.; Tran, T.L.; Kim, W.S.; Sung, J.H. Wound-healing effect of ginsenoside Rd from leaves of Panax ginseng via cyclic AMP-dependent protein kinase pathway. Eur. J. Pharmacol. 2013, 702, 285–293. [Google Scholar] [CrossRef] [PubMed]
- Maquart, F.X.; Chastang, F.; Simeon, A.; Birembaut, P.; Gillery, P.; Wegrowski, Y. Triterpenes from Centella asiatica stimulate extracellular matrix accumulation in rat experimental wounds. Eur. J. Dermatol. 1999, 9, 289–296. [Google Scholar] [PubMed]
- Phaechamud, T.; Yodkhum, K.; Charoenteeraboon, J.; Tabata, Y. Chitosan-aluminum monostearate composite sponge dressing containing asiaticoside for wound healing and angiogenesis promotion in chronic wound. Mater. Sci. Eng. C Mater. Biol. Appl. 2015, 50, 210–225. [Google Scholar] [CrossRef]
- Li, J.; Bao, G.; E, A.L.; Ding, J.; Li, S.; Sheng, S.; Shen, Z.; Jia, Z.; Lin, C.; Zhang, C.; et al. Betulinic acid enhances the viability of random-pattern skin flaps by activating autophagy. Front. Pharmacol. 2019, 10, 1017. [Google Scholar] [CrossRef] [Green Version]
- Cheng, S.; Zhang, X.; Feng, Q.; Chen, J.; Shen, L.; Yu, P.; Yang, L.; Chen, D.; Zhang, H.; Sun, W.; et al. Astragaloside IV exerts angiogenesis and cardioprotection after myocardial infarction via regulating PTEN/PI3K/Akt signaling pathway. Life Sci. 2019, 227, 82–93. [Google Scholar] [CrossRef]
- Huang, C.; Jacobson, K.; Schaller, M.D. MAP kinases and cell migration. J. Cell Sci. 2004, 117, 4619–4628. [Google Scholar] [CrossRef] [Green Version]
- Bonel-Perez, G.C.; Perez-Jimenez, A.; Gris-Cardenas, I.; Parra-Perez, A.M.; Lupianez, J.A.; Reyes-Zurita, F.J.; Siles, E.; Csuk, R.; Peragon, J.; Rufino-Palomares, E.E. Antiproliferative and pro-apoptotic effect of uvaol in human hepatocarcinoma HepG2 cells by affecting G0/G1 cell cycle arrest, ROS production and AKT/PI3K signaling pathway. Molecules 2020, 25, 4254. [Google Scholar] [CrossRef]
- Ramos-Hryb, A.B.; Cunha, M.P.; Pazini, F.L.; Lieberknecht, V.; Prediger, R.D.S.; Kaster, M.P.; Rodrigues, A.L.S. Ursolic acid affords antidepressant-like effects in mice through the activation of PKA, PKC, CAMK-II and MEK1/2. Pharmacol. Rep. 2017, 69, 1240–1246. [Google Scholar] [CrossRef]
- Wang, Z.; Li, Q.; Xiang, M.; Zhang, F.; Wei, D.; Wen, Z.; Zhou, Y. Astragaloside alleviates hepatic fibrosis function via PAR2 signaling pathway in diabetic rats. Cell. Physiol. Biochem. 2017, 41, 1156–1166. [Google Scholar] [CrossRef]
- Castro, A.J.G.; Cazarolli, L.H.; da Luz, G.; Altenhofen, D.; da Silva, H.B.; de Carvalho, F.K.; Pizzolatti, M.G.; Silva, F. Fern-9(11)-ene-2alpha,3beta-diol Action on insulin secretion under hyperglycemic conditions. Biochemistry 2018, 57, 3894–3902. [Google Scholar] [CrossRef]
- Bodnar, R.J.; Yates, C.C.; Wells, A. IP-10 blocks vascular endothelial growth factor-induced endothelial cell motility and tube formation via inhibition of calpain. Circ. Res. 2006, 98, 617–625. [Google Scholar] [CrossRef]
- Lv, T.; Du, Y.; Cao, N.; Zhang, S.; Gong, Y.; Bai, Y.; Wang, W.; Liu, H. Proliferation in cardiac fibroblasts induced by beta1-adrenoceptor autoantibody and the underlying mechanisms. Sci. Rep. 2016, 6, 32430. [Google Scholar] [CrossRef]
- Ling, G.; Ji, Q.; Ye, W.; Ma, D.; Wang, Y. Epithelial-mesenchymal transition regulated by p38/MAPK signaling pathways participates in vasculogenic mimicry formation in SHG44 cells transfected with TGF-beta cDNA loaded lentivirus in vitro and in vivo. Int. J. Oncol. 2016, 49, 2387–2398. [Google Scholar] [CrossRef] [Green Version]
- Barchitta, M.; Maugeri, A.; Favara, G.; Magnano San Lio, R.; Evola, G.; Agodi, A.; Basile, G. Nutrition and wound healing: an overview focusing on the beneficial effects of curcumin. Int. J. Mol. Sci. 2019, 20, 1119. [Google Scholar] [CrossRef] [Green Version]
- Landen, N.X.; Li, D.; Stahle, M. Transition from inflammation to proliferation: A critical step during wound healing. Cell. Mol. Life Sci. 2016, 73, 3861–3885. [Google Scholar] [CrossRef] [Green Version]
- Du, S.Y.; Huang, H.F.; Li, X.Q.; Zhai, L.X.; Zhu, Q.C.; Zheng, K.; Song, X.; Xu, C.S.; Li, C.Y.; Li, Y.; et al. Anti-inflammatory properties of uvaol on DSS-induced colitis and LPS-stimulated macrophages. Chin. Med. 2020, 15, 43. [Google Scholar] [CrossRef]
- Nguyen, V.L.; Truong, C.T.; Nguyen, B.C.Q.; Vo, T.V.; Dao, T.T.; Nguyen, V.D.; Trinh, D.T.; Huynh, H.K.; Bui, C.B. Anti-inflammatory and wound healing activities of calophyllolide isolated from Calophyllum inophyllum Linn. PLoS ONE 2017, 12, e0185674. [Google Scholar] [CrossRef] [Green Version]
- Da Silva, E.C.O.; Dos Santos, F.M.; Ribeiro, A.R.B.; de Souza, S.T.; Barreto, E.; Fonseca, E. Drug-induced anti-inflammatory response in A549 cells, as detected by Raman spectroscopy: A comparative analysis of the actions of dexamethasone and p-coumaric acid. Analyst 2019, 144, 1622–1631. [Google Scholar] [CrossRef]
- Cardoso, S.H.; de Oliveira, C.R.; Guimaraes, A.S.; Nascimento, J.; de Oliveira Dos Santos Carmo, J.; de Souza Ferro, J.N.; de Carvalho Correia, A.C.; Barreto, E. Synthesis of newly functionalized 1,4-naphthoquinone derivatives and their effects on wound healing in alloxan-induced diabetic mice. Chem. Biol. Interact. 2018, 291, 55–64. [Google Scholar] [CrossRef]
- Davis, P.K.; Ho, A.; Dowdy, S.F. Biological methods for cell-cycle synchronization of mammalian cells. Biotechniques 2001, 30, 1322–1331. [Google Scholar] [CrossRef] [Green Version]
- Smaniotto, S.; de Mello-Coelho, V.; Villa-Verde, D.M.; Pleau, J.M.; Postel-Vinay, M.C.; Dardenne, M.; Savino, W. Growth hormone modulates thymocyte development in vivo through a combined action of laminin and CXC chemokine ligand 12. Endocrinology 2005, 146, 3005–3017. [Google Scholar] [CrossRef] [Green Version]
- Dong, Z.; Zhao, X.; Tai, W.; Lei, W.; Wang, Y.; Li, Z.; Zhang, T. IL-27 attenuates the TGF-beta1-induced proliferation, differentiation and collagen synthesis in lung fibroblasts. Life Sci. 2016, 146, 24–33. [Google Scholar] [CrossRef] [PubMed]
- Agra, I.; Pires, L.L.; Carvalho, P.S.; Silva-Filho, E.A.; Smaniotto, S.; Barreto, E. Evaluation of wound healing and antimicrobial properties of aqueous extract from Bowdichia virgilioides stem barks in mice. An. Acad. Bras. Cienc. 2013, 85, 945–954. [Google Scholar] [CrossRef]
Sample Availability: Samples of the compounds are not available from the authors. |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
Share and Cite
Carmo, J.; Cavalcante-Araújo, P.; Silva, J.; Ferro, J.; Correia, A.C.; Lagente, V.; Barreto, E. Uvaol Improves the Functioning of Fibroblasts and Endothelial Cells and Accelerates the Healing of Cutaneous Wounds in Mice. Molecules 2020, 25, 4982. https://doi.org/10.3390/molecules25214982
Carmo J, Cavalcante-Araújo P, Silva J, Ferro J, Correia AC, Lagente V, Barreto E. Uvaol Improves the Functioning of Fibroblasts and Endothelial Cells and Accelerates the Healing of Cutaneous Wounds in Mice. Molecules. 2020; 25(21):4982. https://doi.org/10.3390/molecules25214982
Chicago/Turabian StyleCarmo, Julianderson, Polliane Cavalcante-Araújo, Juliane Silva, Jamylle Ferro, Ana Carolina Correia, Vincent Lagente, and Emiliano Barreto. 2020. "Uvaol Improves the Functioning of Fibroblasts and Endothelial Cells and Accelerates the Healing of Cutaneous Wounds in Mice" Molecules 25, no. 21: 4982. https://doi.org/10.3390/molecules25214982