Protective Effects of Withagenin A Diglucoside from Indian Ginseng (Withania somnifera) against Human Dermal Fibroblast Damaged by TNF-α Stimulation
Abstract
:1. Introduction
2. Materials and Methods
2.1. General Experimental Procedure and Plant Material
2.2. Extraction and Separation of WAD
2.3. Cell Culture Condition
2.4. Sample Preparation
2.5. Cell Viability Assay
2.6. Intracellular ROS Generation Assay
2.7. ELISA Assay
2.8. Immunoblotting Assay
2.9. Statistical Analyses
3. Results
3.1. Isolation and Identification of Withagenin A Diglucoside (WAD)
3.2. Effect of WAD on Viability of HDFs
3.3. Effect of WAD on Intracellular ROS (Reactive Oxygen Species) Secretion in HDFs
3.4. Effect of WAD on MMP-1 and COLIA1 Protein Secretion in HDFs
3.5. Effect of WAD on Phosphorylation of MAPK Protein Expression in HDFs
3.6. Effect of WAD on Phosphorylation of c-Jun and NF-κB Protein Expression in HDFs
3.7. Effect of WAD on Phosphorylation of Akt, COX-2, and HO-1 in HDFs
3.8. Effect of WAD on IL-6 and IL-8 Secretion in HDFs
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
- Woo, W.M. Skin Structure and Biology. In Imaging Technologies and Transdermal Delivery in Skin Disorders; Wiley-VCH: Weinheim, Germany, 2019; pp. 1–14. [Google Scholar]
- Thankam, F.G.; Dilisio, M.F.; Gross, R.M.; Agrawal, D.K. Collagen I: A kingpin for rotator cuff tendon pathology. Am. J. Transl. Res. 2018, 10, 3291–3309. [Google Scholar]
- Zhang, S.; Duan, E. Fighting against skin aging: The way from bench to bedside. Cell Transplant. 2018, 27, 729–738. [Google Scholar] [CrossRef] [PubMed]
- Huertas, A.C.M.; Schmelzer, C.E.; Hoehenwarter, W.; Heyroth, F.; Heinz, A. Molecular-level insights into aging processes of skin elastin. Biochimie 2016, 128, 163–173. [Google Scholar] [CrossRef] [PubMed]
- McDaniel, D.; Farris, P.; Valacchi, G. Atmospheric skin aging—Contributors and inhibitors. J. Cosmet. Dermatol. 2018, 17, 124–137. [Google Scholar] [CrossRef] [PubMed]
- Choi, J.W.; Lee, J.; Park, Y.I. 7, 8-Dihydroxyflavone attenuates TNF-α-induced skin aging in Hs68 human dermal fibroblast cells via down-regulation of the MAPKs/Akt signaling pathways. Biomed. Pharmacother. 2017, 95, 1580–1587. [Google Scholar] [CrossRef] [PubMed]
- Phung, H.M.; Lee, S.; Hong, S.; Lee, S.; Jung, K.; Kang, K.S. Protective Effect of Polymethoxyflavones Isolated from Kaempferia parviflora against TNF-α-Induced Human Dermal Fibroblast Damage. Antioxidants 2021, 10, 1609. [Google Scholar] [CrossRef]
- Lee, S.; Yu, J.S.; Phung, H.M.; Lee, J.G.; Kim, K.H.; Kang, K.S. Potential anti-skin aging effect of (-)-Catechin isolated from the root bark of ulmus davidiana var. japonica in tumor necrosis factor-α-stimulated normal human dermal fibroblasts. Antioxidants 2020, 9, 981. [Google Scholar] [CrossRef]
- Lee, S.; Jang, T.; Kim, K.H.; Kang, K.S. Improvement of Damage in Human Dermal Fibroblasts by 3, 5, 7-Trimethoxyflavone from Black Ginger (Kaempferia parviflora). Antioxidants 2022, 11, 425. [Google Scholar] [CrossRef]
- Kulkarni, S.; Dhir, A. Withania somnifera: An Indian ginseng. Prog. Neuro-Psychopharmacol. Biol. Psychiatry 2008, 32, 1093–1105. [Google Scholar] [CrossRef]
- Pandey, V.; Ansari, W.A.; Misra, P.; Atri, N. Withania somnifera: Advances and implementation of molecular and tissue culture techniques to enhance its application. Front. Plant Sci. 2017, 8, 1390. [Google Scholar] [CrossRef] [Green Version]
- Chakraborti, S.; de Barun, K.; Bandyopadhyay, T. Variations in the antitumour constituents of Withania somnifera dunal. Experientia 1974, 30, 852–853. [Google Scholar] [CrossRef]
- Minguzzi, S.; Barata, L.E.; Shin, Y.G.; Jonas, P.F.; Chai, H.-B.; Park, E.J.; Pezzuto, J.M.; Cordell, G.A. Cytotoxic withanolides from Acnistus arborescens. Phytochemistry 2002, 59, 635–641. [Google Scholar] [CrossRef]
- Zhao, J.; Nakamura, N.; Hattori, M.; Kuboyama, T.; Tohda, C.; Komatsu, K. Withanolide derivatives from the roots of Withania somnifera and their neurite outgrowth activities. Chem. Pharm. Bull. 2002, 50, 760–765. [Google Scholar] [CrossRef] [Green Version]
- Zhang, H.; Cao, C.-M.; Gallagher, R.J.; Timmermann, B.N. Antiproliferative withanolides from several solanaceous species. Nat. Prod. Res. 2014, 28, 1941–1951. [Google Scholar] [CrossRef] [Green Version]
- Ma, T.; Zhang, W.-N.; Yang, L.; Zhang, C.; Lin, R.; Shan, S.-M.; Zhu, M.-D.; Luo, J.-G.; Kong, L.-Y. Cytotoxic withanolides from Physalis angulata var. villosa and the apoptosis-inducing effect via ROS generation and the activation of MAPK in human osteosarcoma cells. RSC Adv. 2016, 6, 53089–53100. [Google Scholar] [CrossRef]
- Tomar, V.; Beuerle, T.; Sircar, D. A validated HPTLC method for the simultaneous quantifications of three phenolic acids and three withanolides from Withania somnifera plants and its herbal products. J. Chromatogr. B 2019, 1124, 154–160. [Google Scholar] [CrossRef]
- Kim, S.; Yu, J.S.; Lee, J.Y.; Choi, S.U.; Lee, J.; Kim, K.H. Cytotoxic withanolides from the roots of Indian ginseng (Withania somnifera). J. Nat. Prod. 2019, 82, 765–773. [Google Scholar] [CrossRef]
- Lee, B.S.; Yoo, M.J.; Kang, H.; Lee, S.R.; Kim, S.; Yu, J.S.; Kim, J.-C.; Jang, T.S.; Pang, C.; Kim, K.H. Withasomniferol D, a New Anti-Adipogenic Withanolide from the Roots of Ashwagandha (Withania somnifera). Pharmaceuticals 2021, 14, 1017. [Google Scholar] [CrossRef]
- Chikhale, R.V.; Gurav, S.S.; Patil, R.B.; Sinha, S.K.; Prasad, S.K.; Shakya, A.; Shrivastava, S.K.; Gurav, N.S.; Prasad, R.S. Sars-cov-2 host entry and replication inhibitors from Indian ginseng: An in-silico approach. J. Biomol. Struct. Dyn. 2021, 39, 4510–4521. [Google Scholar] [CrossRef]
- Tripathi, M.K.; Singh, P.; Sharma, S.; Singh, T.P.; Ethayathulla, A.; Kaur, P. Identification of bioactive molecule from Withania somnifera (Ashwagandha) as SARS-CoV-2 main protease inhibitor. J. Biomol. Struct. Dyn. 2021, 39, 5668–5681. [Google Scholar] [CrossRef]
- Ha, J.W.; Yu, J.S.; Lee, B.S.; Kang, D.-M.; Ahn, M.-J.; Kim, J.K.; Kim, K.H. Structural Characterization of Withanolide Glycosides from the Roots of Withania somnifera and Their Potential Biological Activities. Plants 2022, 11, 767. [Google Scholar] [CrossRef] [PubMed]
- Lee, S.R.; Kang, H.; Yoo, M.J.; Yu, J.S.; Lee, S.; Yi, S.A.; Beemelmanns, C.; Lee, J.; Kim, K.H. Anti-adipogenic pregnane steroid from a Hydractinia-associated fungus, Cladosporium sphaerospermum SW67. Nat. Prod. Sci. 2020, 26, 230–235. [Google Scholar]
- Lee, S.; Ryoo, R.; Choi, J.H.; Kim, J.-H.; Kim, S.-H.; Kim, K.H. Trichothecene and tremulane sesquiterpenes from a hallucinogenic mushroom Gymnopilus junonius and their cytotoxicity. Arch. Pharm. Res. 2020, 43, 214–223. [Google Scholar] [CrossRef]
- Ha, J.W.; Kim, J.; Kim, H.; Jang, W.; Kim, K.H. Mushrooms: An important source of natural bioactive compounds. Nat. Prod. Sci. 2020, 26, 118–131. [Google Scholar]
- Lee, S.; Kim, C.S.; Yu, J.S.; Kang, H.; Yoo, M.J.; Youn, U.J.; Ryoo, R.; Bae, H.Y.; Kim, K.H. Ergopyrone, a Styrylpyrone-Fused Steroid with a Hexacyclic 6/5/6/6/6/5 Skeleton from a Mushroom Gymnopilus orientispectabilis. Org. Lett. 2021, 23, 3315–3319. [Google Scholar] [CrossRef]
- Lee, K.H.; Kim, J.K.; Yu, J.S.; Jeong, S.Y.; Choi, J.H.; Kim, J.-C.; Ko, Y.-J.; Kim, S.-H.; Kim, K.H. Ginkwanghols A and B, osteogenic coumaric acid-aliphatic alcohol hybrids from the leaves of Ginkgo biloba. Arch. Pharm. Res. 2021, 44, 514–524. [Google Scholar] [CrossRef]
- Bolleddula, J.; Fitch, W.; Vareed, S.K.; Nair, M.G. Identification of metabolites in Withania sominfera fruits by liquid chromatography and high-resolution mass spectrometry. Rapid Commun. Mass Spectrom. 2012, 26, 1277–1290. [Google Scholar] [CrossRef]
- Panchenko, M.V.; Farber, H.W.; Korn, J.H. Induction of heme oxygenase-1 by hypoxia and free radicals in human dermal fibroblasts. Am. J. Physiol. Cell Physiol. 2000, 278, C92–C101. [Google Scholar] [CrossRef] [Green Version]
- Deshmukh, S.N.; Dive, A.M.; Moharil, R.; Munde, P. Enigmatic insight into collagen. J. Oral Maxillofac. Pathol. JOMFP 2016, 20, 276. [Google Scholar] [CrossRef]
- Stefanovic, B. RNA protein interactions governing expression of the most abundant protein in human body, type I collagen. Wiley Interdiscip. Rev. RNA 2013, 4, 535–545. [Google Scholar] [CrossRef] [Green Version]
- Kim, S.-J.; Park, J.-H.; Kim, D.-H.; Won, Y.-H.; Maibach, H.I. Increased in vivo collagen synthesis and in vitro cell proliferative effect of glycolic acid. Dermatol. Surg. 1998, 24, 1054–1058. [Google Scholar] [CrossRef]
- Chavoshnejad, P.; Foroughi, A.H.; Dhandapani, N.; German, G.K.; Razavi, M.J. Effect of collagen degradation on the mechanical behavior and wrinkling of skin. Phys. Rev. E 2021, 104, 034406. [Google Scholar] [CrossRef]
- Lorenz, W.; Buhrmann, C.; Mobasheri, A.; Lueders, C.; Shakibaei, M. Bacterial lipopolysaccharides form procollagen-endotoxin complexes that trigger cartilage inflammation and degeneration: Implications for the development of rheumatoid arthritis. Arthritis Res. Ther. 2013, 15, R111. [Google Scholar] [CrossRef]
- Cleutjens, J.P. The role of matrix metalloproteinases in heart disease. Cardiovasc. Res. 1996, 32, 816–821. [Google Scholar] [CrossRef]
- Parrado, C.; Mercado-Saenz, S.; Perez-Davó, A.; Gilaberte, Y.; Gonzalez, S.; Juarranz, A. Environmental stressors on skin aging. Mechanistic insights. Front. Pharmacol. 2019, 10, 759. [Google Scholar] [CrossRef]
- Misra, B.B. The chemical exposome of human aging. Front. Genet. 2020, 11, 574936. [Google Scholar] [CrossRef]
- Bosch, R.; Philips, N.; Suárez-Pérez, J.A.; Juarranz, A.; Devmurari, A.; Chalensouk-Khaosaat, J.; González, S. Mechanisms of photoaging and cutaneous photocarcinogenesis, and photoprotective strategies with phytochemicals. Antioxidants 2015, 4, 248–268. [Google Scholar] [CrossRef] [Green Version]
- Madan, K.; Nanda, S. In-vitro evaluation of antioxidant, anti-elastase, anti-collagenase, anti-hyaluronidase activities of safranal and determination of its sun protection factor in skin photoaging. Bioorg. Chem. 2018, 77, 159–167. [Google Scholar] [CrossRef]
- Shin, J.-W.; Kwon, S.-H.; Choi, J.-Y.; Na, J.-I.; Huh, C.-H.; Choi, H.-R.; Park, K.-C. Molecular mechanisms of dermal aging and antiaging approaches. Int. J. Mol. Sci. 2019, 20, 2126. [Google Scholar] [CrossRef] [Green Version]
- Philips, N.; Keller, T.; Hendrix, C.; Hamilton, S.; Arena, R.; Tuason, M.; Gonzalez, S. Regulation of the extracellular matrix remodeling by lutein in dermal fibroblasts, melanoma cells, and ultraviolet radiation exposed fibroblasts. Arch. Dermatol. Res. 2007, 299, 373–379. [Google Scholar] [CrossRef]
- Fligiel, S.E.; Varani, J.; Datta, S.C.; Kang, S.; Fisher, G.J.; Voorhees, J.J. Collagen degradation in aged/photodamaged skin in vivo and after exposure to matrix metalloproteinase-1 in vitro. J. Investig. Dermatol. 2003, 120, 842–848. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Kammeyer, A.; Luiten, R. Oxidation events and skin aging. Ageing Res. Rev. 2015, 21, 16–29. [Google Scholar] [CrossRef]
- Kang, M.; Park, S.-H.; Park, S.J.; Oh, S.W.; Yoo, J.A.; Kwon, K.; Kim, J.; Yu, E.; Cho, J.Y.; Lee, J. p44/42 MAPK signaling is a prime target activated by phenylethyl resorcinol in its anti-melanogenic action. Phytomedicine 2019, 58, 152877. [Google Scholar] [CrossRef] [PubMed]
- Ansary, T.M.; Hossain, M.R.; Kamiya, K.; Komine, M.; Ohtsuki, M. Inflammatory molecules associated with ultraviolet radiation-mediated skin aging. Int. J. Mol. Sci. 2021, 22, 3974. [Google Scholar] [CrossRef] [PubMed]
- Bernard, J.J.; Gallo, R.L.; Krutmann, J. Photoimmunology: How ultraviolet radiation affects the immune system. Nat. Rev. Immunol. 2019, 19, 688–701. [Google Scholar] [CrossRef]
- Cavinato, M.; Koziel, R.; Romani, N.; Weinmüllner, R.; Jenewein, B.; Hermann, M.; Dubrac, S.; Ratzinger, G.; Grillari, J.; Schmuth, M.; et al. UVB-induced senescence of human dermal fibroblasts involves impairment of proteasome and enhanced autophagic activity. J. Gerontol. Ser. A Biomed. Sci. Med. Sci. 2017, 72, 632–639. [Google Scholar] [CrossRef] [Green Version]
- Jung, Y.R.; Kim, D.H.; Kim, S.R.; An, H.J.; Lee, E.K.; Tanaka, T.; Kim, N.D.; Yokozawa, T.; Park, J.N.; Chung, H.Y. Anti-wrinkle effect of magnesium lithospermate B from Salvia miltiorrhiza BUNGE: Inhibition of MMPs via NF-kB signaling. PLoS ONE 2014, 9, e102689. [Google Scholar] [CrossRef] [Green Version]
- Silvers, A.L.; Bachelor, M.A.; Bowden, G.T. The role of JNK and p38 MAPK activities in UVA-induced signaling pathways leading to AP-1 activation and c-Fos expression. Neoplasia 2003, 5, 319–329. [Google Scholar] [CrossRef] [Green Version]
- Kim, M.; Park, H.J. Molecular Mechanisms of Skin Aging and Rejuvenation. In Molecular Mechanisms of the Aging Process and Rejuvenation; Shiomi, N., Ed.; IntechOpen: London, UK, 2016; p. 450. [Google Scholar]
- Palungwachira, P.; Tancharoen, S.; Phruksaniyom, C.; Klungsaeng, S.; Srichan, R.; Kikuchi, K.; Nararatwanchai, T. Antioxidant and anti-inflammatory properties of anthocyanins extracted from Oryza sativa L. in primary dermal fibroblasts. Oxidative Med. Cell. Longev. 2019, 2019, 2089817. [Google Scholar] [CrossRef] [Green Version]
- Bang, E.; Kim, D.H.; Chung, H.Y. Protease-activated receptor 2 induces ROS-mediated inflammation through Akt-mediated NF-κB and FoxO6 modulation during skin photoaging. Redox Biol. 2021, 44, 102022. [Google Scholar] [CrossRef]
- Choi, Y.J.; Moon, K.M.; Chung, K.W.; Jeong, J.W.; Park, D.; Kim, D.H.; Yu, B.P.; Chung, H.Y. The underlying mechanism of proinflammatory NF-κB activation by the mTORC2/Akt/IKKα pathway during skin aging. Oncotarget 2016, 7, 52685–52694. [Google Scholar] [CrossRef]
- Choi, K.-S.; Kundu, J.K.; Chun, K.-S.; Na, H.-K.; Surh, Y.-J. Rutin inhibits UVB radiation-induced expression of COX-2 and iNOS in hairless mouse skin: p38 MAP kinase and JNK as potential targets. Arch. Biochem. Biophys. 2014, 559, 38–45. [Google Scholar] [CrossRef]
- Subedi, L.; Lee, T.H.; Wahedi, H.M.; Baek, S.-H.; Kim, S.Y. Resveratrol-enriched rice attenuates UVB-ROS-induced skin aging via downregulation of inflammatory cascades. Oxidative Med. Cell. Longev. 2017, 2017, 8379539. [Google Scholar] [CrossRef]
- Surowiak, P.; Gansukh, T.; Donizy, P.; Halon, A.; Rybak, Z. Increase in cyclooxygenase-2 (COX-2) expression in keratinocytes and dermal fibroblasts in photoaged skin. J. Cosmet. Dermatol. 2014, 13, 195–201. [Google Scholar] [CrossRef]
- Tu, Y.; Quan, T. Oxidative stress and human skin connective tissue aging. Cosmetics 2016, 3, 28. [Google Scholar] [CrossRef] [Green Version]
- Nelson, G.; Kucheryavenko, O.; Wordsworth, J.; von Zglinicki, T. The senescent bystander effect is caused by ROS-activated NF-κB signalling. Mech. Ageing Dev. 2018, 170, 30–36. [Google Scholar] [CrossRef]
- Lingappan, K. NF-κB in oxidative stress. Curr. Opin. Toxicol. 2018, 7, 81–86. [Google Scholar] [CrossRef]
- Kim, H.-R.; Jeong, D.-H.; Kim, S.; Lee, S.-W.; Sin, H.-S.; Yu, K.-Y.; Jeong, S.-I.; Kim, S.-Y. Fermentation of blackberry with L. plantarum JBMI F5 enhance the protection effect on UVB-mediated photoaging in human foreskin fibroblast and hairless mice through regulation of MAPK/NF-κB signaling. Nutrients 2019, 11, 2429. [Google Scholar] [CrossRef] [Green Version]
- Hwang, S.J.; Ha, G.-H.; Seo, W.-Y.; Kim, C.K.; Kim, K.; Lee, S.B. Human collagen alpha-2 type I stimulates collagen synthesis, wound healing, and elastin production in normal human dermal fibroblasts (HDFs). BMB Rep. 2020, 53, 539. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 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 (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Lee, S.; Choi, Y.J.; Lee, S.; Kang, K.S.; Jang, T.S.; Kim, K.H. Protective Effects of Withagenin A Diglucoside from Indian Ginseng (Withania somnifera) against Human Dermal Fibroblast Damaged by TNF-α Stimulation. Antioxidants 2022, 11, 2248. https://doi.org/10.3390/antiox11112248
Lee S, Choi YJ, Lee S, Kang KS, Jang TS, Kim KH. Protective Effects of Withagenin A Diglucoside from Indian Ginseng (Withania somnifera) against Human Dermal Fibroblast Damaged by TNF-α Stimulation. Antioxidants. 2022; 11(11):2248. https://doi.org/10.3390/antiox11112248
Chicago/Turabian StyleLee, Sullim, Yea Jung Choi, Seulah Lee, Ki Sung Kang, Tae Su Jang, and Ki Hyun Kim. 2022. "Protective Effects of Withagenin A Diglucoside from Indian Ginseng (Withania somnifera) against Human Dermal Fibroblast Damaged by TNF-α Stimulation" Antioxidants 11, no. 11: 2248. https://doi.org/10.3390/antiox11112248