Journal of Ginseng Research

The Korean Society of Ginseng (고려인삼학회)
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Panax ginseng total protein promotes proliferation and secretion of collagen in NIH/3T3 cells by activating extracellular signal-related kinase pathway

  • Chen, Xuenan (Research Center of Traditional Chinese Medicine, Affiliated Hospital to Changchun University of Chinese Medicine) ;
  • Wang, Manying (Research Center of Traditional Chinese Medicine, Affiliated Hospital to Changchun University of Chinese Medicine) ;
  • Xu, Xiaohao (Jilin Technology Innovation Center for Chinese Medicine Biotechnology, College of Biology and Chemistry, Beihua University) ;
  • Liu, Jianzeng (Jilin Technology Innovation Center for Chinese Medicine Biotechnology, College of Biology and Chemistry, Beihua University) ;
  • Mei, Bing (Research and Development Center of Traditional Chinese Medicine and Biological Engineering, Changchun University of Chinese Medicine) ;
  • Fu, Pingping (China-Japan Union Hospital and First Affiliated Hospital of Jilin University) ;
  • Zhao, Daqing (Research and Development Center of Traditional Chinese Medicine and Biological Engineering, Changchun University of Chinese Medicine) ;
  • Sun, Liwei (Jilin Technology Innovation Center for Chinese Medicine Biotechnology, College of Biology and Chemistry, Beihua University)
  • Received : 2016.11.09
  • Accepted : 2017.02.13
  • Published : 2017.07.15
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Abstract

Background: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. Methods: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ${\beta}1$, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. Results: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ${\beta}1$ and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. Conclusion: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.

Keywords

References

  1. Attele AS, Wu JA, Yuan CS. Ginseng pharmacology: multiple constituents and multiple actions. Biochem Pharmacol 1999;58:1685-93. PubMed PMID:10571242. https://doi.org/10.1016/S0006-2952(99)00212-9
  2. Chen X. Cardiovascular protection by ginsenosides and their nitric oxide releasing action. Clin Exp Pharmacol Physiol 1996;23:728-32. PubMed PMID:8886498. https://doi.org/10.1111/j.1440-1681.1996.tb01767.x
  3. Nag SA, Qin JJ, Wang W, Wang MH, Wang H, Zhang R. Ginsenosides as anticancer agents: in vitro and in vivo activities, structure-activity relationships, and molecular mechanisms of action. Front Pharmacol 2012;3:25. http://dx.doi.org/10.3389/fphar.2012.00025. PubMed PMID: 22403544; PubMed Central PMCID: PMC3289390.
  4. Kim YG, Sumiyoshi M, Kawahira K, Sakanaka M, Kimura Y. Effects of Red Ginseng extract on ultraviolet B-irradiated skin change in C57BL mice. Phytother Res 2008;22:1423-7. http://dx.doi.org/10.1002/ptr. 2339. PubMed PMID: 18803235.
  5. Kimura Y, Sumiyoshi M, Kawahira K, Sakanaka M. Effects of ginseng saponins isolated from Red Ginseng roots on burn wound healing in mice. Br J Pharmacol 2006;148:860-70. http://dx.doi.org/10.1038/sj.bjp.0706794. PubMed PMID: 16770323; PubMed Central PMCID: PMC1617068.
  6. Ru W, Wang D, Xu Y, He X, Sun YE, Qian L, et al. Chemical constituents and bioactivities of Panax ginseng (C. A. Mey.). Drug Discov Ther 2015;9:23-32. http://dx.doi.org/10.5582/ddt.2015.01004. PubMed PMID: 25788049.
  7. Li H, Kang T, Qi B, Kong L, Jiao Y, Cao Y, et al. Neuroprotective effects of ginseng protein on PI3K/Akt signaling pathway in the hippocampus of d-galactose/$AlCl_3$ inducing rats model of Alzheimer's disease. J Ethnopharmacol 2016;179: 162-9. http://dx.doi.org/10.1016/j.jep.2015.12.020. PubMed PMID:26721223.
  8. Li H, Song J, Zhang J, Wang T, Yan Y, Tao Z, et al. Ginseng protein reverses amyloid beta peptide and $H_2O_2$ cytotoxicity in neurons, and ameliorates cognitive impairment in AD rats induced by a combination of d-galactose and $AlCl_3$. Phytother Res 2017;31:284-95. http://dx.doi.org/10.1002/ptr. 5747. PubMed PMID: 27981642.
  9. Ng TB, Wang H. Panaxagin, a new protein from Chinese ginseng possesses anti-fungal, anti-viral, translation-inhibiting and ribonuclease activities. Life Sci 2001;68:739-49. PubMed PMID: 11205866. https://doi.org/10.1016/S0024-3205(00)00970-X
  10. Qi B, Wang S, Wang Q, Zhang H, Bai XY, et al. Characterization and immunostimulating effects on murine peritoneal macrophages of a novel protein isolated from Panax quinquefolius L. J Ethnopharmacol 2016;193:700-5. http://dx.doi.org/10.1016/j.jep.2016.10.034. PubMed PMID: 27742408.
  11. Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev 2003;83:835-70. http://dx.doi.org/10.1152/physrev. 00031.2002. PubMed PMID: 12843410.
  12. Gordon MK, Hahn RA. Collagens. Cell Tissue Res 2010;339:247-57. http://dx.doi.org/10.1007/s00441-009-0844-4. PubMed PMID: 19693541; PubMed Central PMCID: PMC2997103.
  13. Bao P, Kodra A, Tomic-Canic M, Golinko MS, Ehrlich HP, Brem H. The role of vascular endothelial growth factor in wound healing. J Surg Res 2009;153:347-58. http://dx.doi.org/10.1016/j.jss.2008.04.023. PubMed PMID: 19027922; PubMed Central PMCID: PMC2728016.
  14. Lai CF, Chaudhary L, Fausto A, Halstead LR, Ory DS, Avioli LV, et al. Erk is essential for growth, differentiation, integrin expression, and cell function in human osteoblastic cells. J Biol Chem 2001;276:14443-50. http://dx.doi.org/10.1074/jbc.M010021200. PubMed PMID: 11278600.
  15. Juretic N, Santibanez JF, Hurtado C, Martinez J. ERK 1,2 and p38 pathways are involved in the proliferative stimuli mediated by urokinase in osteoblastic SaOS-2 cell line. J Cell Biochem 2001;83:92-8. PubMed PMID: 11500957. https://doi.org/10.1002/jcb.1211
  16. Craxton A, Shu G, Graves JD, Saklatvala J, Krebs EG, Clark EA. p38 MAPK is required for CD40-induced gene expression and proliferation in B lymphocytes. J Immunol 1998;161:3225-36. PubMed PMID: 9759836.
  17. Hong NY, Cui ZG, Kang HK, Lee DH, Lee YK, Park DB. p-Synephrine stimulates glucose consumption via AMPK in L6 skeletal muscle cells. Biochem Biophys Res Commun 2012;418:720-4. http://dx.doi.org/10.1016/j.bbrc.2012.01.085.PubMed PMID: 22306011.
  18. Yeom CH, Lee G, Park JH, Yu J, Park S, Yi SY, et al. High dose concentration administration of ascorbic acid inhibits tumor growth in BALB/C mice implanted with sarcoma 180 cancer cells via the restriction of angiogenesis. J Transl Med 2009;7:70. http://dx.doi.org/10.1186/1479-5876-7-70. PubMed PMID: 19671184; PubMed Central PMCID: PMC2732919.
  19. Cui R, Lu Q, Teng Y, Li K, Li N. Chitosan promoted the Corneal epithelial wound healing via activation of ERK pathway. Curr Eye Res 2017;42(1):21-7. http://dx.doi.org/10.3109/02713683.2016.1145235. PubMed PMID: 27259381.
  20. Thornton MJ. Estrogens and aging skin. Dermato-endocrinology 2013;5:264- 70. http://dx.doi.org/10.4161/derm.23872. PubMed PMID: 24194966;PubMed Central PMCID: PMC3772914.
  21. Verdier-Sevrain S, Bonte F, Gilchrest B. Biology of estrogens in skin: implications for skin aging. Exp Dermatol 2006;15:83-94. http://dx.doi.org/10.1111/j.1600-0625.2005.00377.x. PubMed PMID: 16433679.
  22. Liu G, Wang X, Sun X, Deng C, Atala A, Zhang Y. The effect of urine-derived stem cells expressing VEGF loaded in collagen hydrogels on myogenesis and innervation following after subcutaneous implantation in nude mice. Biomaterials 2013;34:8617-29. http://dx.doi.org/10.1016/j.biomaterials.2013.07.077. PubMed PMID: 23932297.
  23. Rosano JM, Cheheltani R, Wang B, Vora H, Kiani MF, Crabbe DL. Targeted delivery of VEGF after a myocardial infarction reduces collagen deposition and improves cardiac function. Cardiovasc Eng Technol 2012;3:237-47. http://dx.doi.org/10.1007/s13239-012-0089-3. PubMed PMID: 22844388; PubMed Central PMCID: PMC3405981.
  24. Nagai N, Kumasaka N, Kawashima T, Kaji H, Nishizawa M, Abe T. Preparation and characterization of collagen microspheres for sustained release of VEGF. J Mater Sci Mater Med 2010;21:1891-8. http://dx.doi.org/10.1007/s10856-010-4054-0. PubMed PMID: 20232232.
  25. Borselli C, Ungaro F, Oliviero O, d'Angelo I, Quaglia F, La Rotonda MI, et al. Bioactivation of collagen matrices through sustained VEGF release from PLGA microspheres. J Biomed Mater Res A 2010;92:94-102. http://dx.doi.org/10.1002/jbm.a.32332. PubMed PMID: 19165799.
  26. Berzal S, Gonzalez-Guerrero C, Rayego-Mateos S, Ucero A, Ocana-Salceda C, Egido J, et al. TNF-related weak inducer of apoptosis (TWEAK) regulates junctional proteins in tubular epithelial cells via canonical NF-kappaB pathway and ERK activation. J Cell Physiol 2015;230:1580-93. http://dx.doi.org/10.1002/jcp.24905. PubMed PMID: 25536182.
  27. Chang HH, Chang MC, Wu IH, Huang GF, Huang WL, Wang YL, et al. Role of ALK5/Smad2/3 and MEK1/ERK signaling in transforming growth factor beta 1-modulated growth, collagen turnover, and differentiation of stem cells from apical papilla of human tooth. J Endod 2015;41:1272-80. http://dx.doi.org/10.1016/j.joen.2015.03.022. PubMed PMID: 26001858.
  28. Cheng S, Guo J, Yang Q, Han L. Crk-like adapter protein is required for TGFbeta-induced AKT and ERK-signaling pathway in epithelial ovarian carcinomas. Tumour Biol 2015;36:915-9. http://dx.doi.org/10.1007/s13277-014-2724-0. PubMed PMID: 25307974.
  29. Muthusubramaniam L, Zaitseva T, Paukshto M, Martin G, Desai T. Effect of collagen nanotopography on keloid fibroblast proliferation and matrix synthesis: implications for dermal wound healing. Tissue Eng Part A 2014;20: 2728-36. http://dx.doi.org/10.1089/ten.TEA.2013.0539. PubMed PMID: 24724556; PubMed Central PMCID: PMC4195479.
  30. Kabeche L, Compton DA. Cyclin A regulates kinetochore microtubules to promote faithful chromosome segregation. Nature 2013;502:110-3. http://dx.doi.org/10.1038/nature12507. PubMed PMID: 24013174; PubMed Central PMCID: PMC3791168.
  31. Ahlin C, Zhou W, Holmqvist M, Holmberg L, Nilsson C, Jirstrom K, et al. Cyclin A is a proliferative marker with good prognostic value in node-negative breast cancer. Cancer Epidemiol Biomarkers Prev 2009;18:2501-6. http://dx.doi.org/10.1158/1055-9965.EPI-09-0169. PubMed PMID: 19706846.
  32. Spiekstra SW, Breetveld M, Rustemeyer T, Scheper RJ, Gibbs S. Wound-healing factors secreted by epidermal keratinocytes and dermal fibroblasts in skin substitutes. Wound Repair Regen 2007;15:708-17. http://dx.doi.org/10.1111/j.1524-475X.2007.00280.x. PubMed PMID: 17971017.
  33. Hunt TK, Burke J, Barbul A, Gimbel ML. Wound healing. Science 1999;284: 1775. PubMed PMID: 10391796.
  34. Witte MB, Barbul A. General principles of wound healing. Surgical Clin North Am 1997;77:509-28. PubMed PMID: 9194878. https://doi.org/10.1016/S0039-6109(05)70566-1
  35. Kane CJ, Hebda PA, Mansbridge JN, Hanawalt PC. Direct evidence for spatial and temporal regulation of transforming growth factor beta 1 expression during cutaneous wound healing. J Cell Physiol 1991;148:157-73. http://dx.doi.org/10.1002/jcp.1041480119. PubMed PMID: 1907288.
  36. Oliver N, Sternlicht M, Gerritsen K, Goldschmeding R. Could aging human skin use a connective tissue growth factor boost to increase collagen content? J Invest Dermatol 2010;130:338-41. http://dx.doi.org/10.1038/jid.2009.331.PubMed PMID: 20081886.
  37. Figueiredo A, Cordeiro AL, Tomada N, Tomada I, Rodrigues A, Gouveia A, et al. Real-time PCR study of Ang1, Ang2, Tie-2, VEGF, and KDR expression in human erectile tissue during aging. J Sexual Med 2011;8:1341-51. http://dx.doi.org/10.1111/j.1743-6109.2010.02116.x. PubMed PMID: 21091880.
  38. Muche A, Bigl M, Arendt T, Schliebs R. Expression of vascular endothelial growth factor (VEGF) mRNA, VEGF receptor 2 (Flk-1) mRNA, and of VEGF coreceptor neuropilin (Nrp)-1 mRNA in brain tissue of aging Tg2576 mice by in situ hybridization. Int J Dev Neurosci 2015;43:25-34. http://dx.doi.org/10.1016/j.ijdevneu.2015.03.003. PubMed PMID: 25797338.
  39. Sebolt-Leopold JS, Herrera R. Targeting the mitogen-activated protein kinase cascade to treat cancer. Nat Rev Cancer 2004;4:937-47. http://dx.doi.org/10.1038/nrc1503. PubMed PMID: 15573115.
  40. Chambard JC, Lefloch R, Pouyssegur J, Lenormand P. ERK implication in cell cycle regulation. Biochim Biophys Acta 2007;1773:1299-310. http://dx.doi.org/10.1016/j.bbamcr.2006.11.010. PubMed PMID: 17188374.
  41. Tamemoto H, Kadowaki T, Tobe K, Ueki K, Izumi T, Chatani Y, et al. Biphasic activation of two mitogen-activated protein kinases during the cell cycle in mammalian cells. J Biol Chem 1992;267:20293-7. PubMed PMID:1400347.
  42. Szoka L, Karna E, Palka J. The mechanism of oxythiamine-induced collagen biosynthesis in cultured fibroblasts. Mol Cell Biochem 2015;403:51-60. http://dx.doi.org/10.1007/s11010-015-2336-z. PubMed PMID: 25626895;PubMed Central PMCID: PMC4383821.

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