20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginsenoside Rb1, enhances the production of hyaluronic acid through the activation of ERK and Akt mediated by Src tyrosin kinase in human keratinocytes

  • Authors:
    • Tae-Gyu Lim
    • Ae Ji Jeon
    • Ji Hye Yoon
    • Dasom Song
    • Jong-Eun Kim
    • Jung Yeon Kwon
    • Jong Rhan Kim
    • Nam Joo Kang
    • Jun-Seong Park
    • Myeong Hun Yeom
    • Deok-Kun Oh
    • Yoongho Lim
    • Charles C. Lee
    • Chang Yong Lee
    • Ki Won Lee
  • View Affiliations

  • Published online on: March 2, 2015     https://doi.org/10.3892/ijmm.2015.2121
  • Pages: 1388-1394
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The aim of the present study was to determine the mechanisms through which 20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol (20GPPD) promotes the production of hyaluronic acid (HA) in human keratinocytes. 20GPPD is the primary bioactive metabolite of Rb1, a major ginsenoside found in ginseng (Panax ginseng). We sought to elucidate the underlying mechanisms behind the 20GPPD-induced production of HA. We found that 20GPPD induced an increase in HA production by elevating hyaluronan synthase 2 (HAS2) expression in human keratinocytes. The phosphorylation of extracellular signal-regulated kinase (ERK) and Akt was also enhanced by 20GPPD in a dose-dependent manner. The pharmacological inhibition of ERK (using U0126) or Akt (using LY294002) suppressed the 20GPPD-induced expression of HAS2, whereas treatment with an epidermal growth factor receptor (EGFR) inhibitor (AG1478) or an intracellular Ca2+ chelator (BAPTA/AM) did not exert any observable effects. The increased Src phosphorylation was also confirmed following treatment with 20GPPD in the human keratinocytes. Following pre-treatment with the Src inhibitor, PP2, both HA production and HAS2 expression were attenuated. Furthermore, the 20GPPD-enhanced ERK and Akt signaling decreased following treatment with PP2. Taken together, our results suggest that Src kinase plays a critical role in the 20GPPD-induced production of HA by acting as an upstream modulator of ERK and Akt activity in human keratinocytes.

Introduction

Ginseng (Panax ginseng) has historically been used as an herbal medicine throughout Asia and is now commonly accessible worldwide. Following ingestion, a bioactive component of ginseng known as ginsenoside Rb1 is converted into 20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol (20GPPD) (Fig. 1A) by gastric fluids and the intestinal microflora (1). Previous studies have demonstrated the beneficial effects of ginsenosides and their metabolites on skin health, such as their wound healing (2,3) and anti-aging effects (4,5), as well as protective effects against untraviolet irradiation (68). Pharmacologically, 20GPPD has anti-inflammatory properties (9), anti-aging effects and promotes skin wound healing (5,10). Previously, our group demonstrated that 20GPPD exerts anticancer effects by inducing colon cancer cell apoptosis (11). It has also been reported that 20GPPD increases hyaluronan synthase (HAS)2 expression in human keratinocytes, although the mechanisms responsible are not yet fully understood.

Hyaluronic acid (HA) is a glycosaminoglycan composed of D-glucuronic acid and N-acetyl-D-glucosamine and functions as a major component of the vitreous body, joint fluids (12), and skin (13). HA plays a role in the proliferation, differentiation and migration of specific cell types (14), and functions through an interaction with cell-surface receptors (15). Approximately 7–8 g of HA are present in an average human body, and half of this amount is found in the skin. HA has the capacity to retain large amounts of water, and thus plays an important role in regulating physiological water balance and osmotic pressure (16). HA content within skin decreases with age (17), and therefore, the upregulation of HA levels may help maintain skin homeostasis during aging.

HA is synthesized in the plasma membrane by HAS1, 2 and 3 (18). Each synthase has distinct encoding sites, producing HA structures of different lengths and with distribution in different tissues during mouse development (19,20). In humans, HAS2 is present in normal tissues, while HAS3 is more commonly expressed in tumor cells or during inflammation (21). The genetic ablation of HAS2 causes embryonic lethality in mouse models, whereas the knockdown of HAS1 and HAS3 knockouts does lead to the development of any observable phenotype (19). In aged human skin, HAS2 gene expression is frequently decreased in the epidermis and dermis (17).

HAS2 is regulated by growth factors and cytokines. Epidermal growth factor (EGF) induces HA synthesis by increasing HAS2 and HAS3 expression (22), while platelet-derived growth factor (PDGF)-BB stimulates HAS2 expression through the activation of phosphatidylinositol 3-kinase (PI3K) and mitogen-activated/extracellular signal-regulated kinase (MEK) (23). Src is a non-receptor tyrosine kinase that plays critical roles in receptor signaling and cellular communication (24). Src is expressed in many cell types, including skin cells, and activates cell proliferation and migration (25). Src is activated by cell adhesion to the extracellular matrix (ECM) and growth factors (26), and subsequently regulates the activation of extracellular signal-regulated kinase (ERK), mitogen-activated protein kinases (MAPK) and PI3K signaling (24), which play a critical role in the PDGF-BB-induced production of HA in human dermal fibroblasts (23).

In the present study, we demonstrate that 20GPPD induces the production of HA through the stimulation of the Src/ERK and Akt signaling pathways in human keratinocytes.

Materials and methods

Materials

20GPPD was purchased from the Ambo Institute (Daejeon, Korea). Dulbecco’s modified Eagle’s medium (DMEM) was obtained from HyClone (Logan, UT, USA). Fetal bovine serum (FBS) and anti-β-actin antibody were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against HAS2 (sc-365263), phosphorylated ERK1/2 (E-4; sc-7383), and total ERK1 (K-23; sc-94) were obtained from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). Antibodies against phosphorylated Src (Tyr416; 2101), Akt (Ser473; 9271), and total Akt (9272) were purchased from Cell Signaling Technology (Beverly, MA, USA). U0126 (ERK inhibitor) was obtained from Tocris Bioscience (Ellisville, MO, USA). LY294002 (Akt inhibitor) was purchased from Cayman Chemical Co. (Ann Arbor, MI, USA). AG1478 (EGFR inhibitor), BAPTA-AM (intracellular Ca2+ chelator) and PP2 (Src inhibitor) were purchased from Calbiochem (Darmstadt, Germany). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) powder was purchased from USB Corp. (Cleveland, OH, USA). Penicillin/streptomycin mix was purchased from Gibco-BRL (Grand Island, NY, USA). The protein assay kit was obtained from Bio-Rad Laboratories (Hercules, CA, USA).

Cell culture and MTT assay

Human keratinocytes (HaCaT; kindly provided by Dr Zigang Dong, Hormel Institute, Univerisity of Minnesota, Minneapolis, MN, USA) were cultured at 37°C in a 5% CO2 atmosphere in DMEM supplemented with 10% FBS, 2 mM L-glutamine and penicillin/streptomycin. Cell cytotoxicity was measured by MTT assay. The cells were cultured in 96-well plates at a density of 2×103 cells/well, and incubated at 37°C in a 5% CO2 atmosphere prior to serum deprivation for 24 h. Various concentrations of 20GPPD were added to the wells for 22 h. Following 2 h of incubation with 20 μl of MTT solution, the medium was removed. Dimethylsulfoxide (DMSO; 200 μl) was added to each well to dissolve the formazan crystals. Absorbance at 570 nm was measured using a microplate reader (Molecular Devices, Sunnyvale, CA, USA).

Enzyme-linked immunosorbent assay (ELISA)

The HaCaT cells were grown to 50% confluence in 6-well plates in DMEM supplemented with 10% FBS. After 24 h of starvation with serum-free DMEM, the medium was removed. The wells were washed with serum-free DMEM to completely remove the HA that had accumulated during cell growth. Among the 6 wells, 2 wells were selected for PP2 pre-treatment for 1 h and the cells were then treated with the indicated concentrations of 20GPPD. The other 3 wells were treated with 20GPPD at various concentrations without PP2 pre-treatment. Following 3 h of incubation, the medium was collected and centrifuged at 1,100 × g for 3 min. The HA concentration in the supernatant was analyzed using an ELISA kit (Corgenix, Inc., Broomfield, CO, USA). ELISA was performed as per the manufacturer’s instructions.

Immunoblot analysis

HaCaT cells were cultured at a density of 2×103 cells for 48 h, prior to serum deprivation for an additional 24 h. The cells were treated with 20GPPD at the indicated concentrations. In order to determine the effects of 20GPPD on HAS2 expression, the cells were incubated with 20GPPD for 3 h. The effect of 20GPPD on ERK, Akt and Src phosphorylation was evaluated after 1 h of 20GPPD treatment. The cells were collected using lysis buffer [10 mM Tris (pH 7.5), 150 mM NaCl, 5 mM EDTA, 1% Triton X-100, 1 mM dithiothreitol (DTT), 0.1 mM phenylmethylsulfonyl fluoride (PMSF), 10% glycerol and a protease inhibitor cocktail tablet] which was prepared manually. Protein concentrations were measured using a dye-binding protein assay kit, according to the manufacturer’s instructions (Bio-Rad Laboratories). Proteins were separated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto PVDF membranes (Millipore Corp., Bedford, MA, USA). The membranes were blocked in 5% skim milk for 2 h and incubated overnight at 4°C with primary antibodies. Subsequently, the membranes were incubated with HRP-conjugated secondary antibodies, and the antibody-bound proteins were detected using a chemiluminescence detection kit (Amersham Pharmacia Biotech, Piscataway, NJ, USA).

Statistical analysis

Data are expressed as the means ± standard deviation (SD), and one-way ANOVA was used for single group comparisons. A value of P<0.05 was used as the criterion for statistical significance.

Results

20GPPD increases HA production in HaCaT cells

A previous study reported that 20GPPD increased HAS2 mRNA expression in HaCaT cells (10). Since HAS2 is a critical enzyme in the production of HA, we sought to investigate the effects of 20GPPD on HA production. The non-cytotoxic concentration range of 20GPPD was first determined (Fig. 1B). Treatment with 20GPPD (0.01–1 μM) increased the production of HA in the HaCaT cells in a dose-dependent manner (Fig. 2A). We further monitored the changes in HAS2 expression in the cells following 1, 3, 6, 9 and 12 h of treatment with 20GPPD. The maximum expression of HAS2 was observed after 3 h of treatment with 20GPPD, and it gradually decreased back to the basal level over the time course of 12 h (Fig. 2B). In accordance with the HA production pattern, the expression of HAS2 was observed to be elevated by 20GPPD in a dose-dependent manner (Fig. 2C). These results demonstrate that 20GPPD induces the production of HA in a dose-dependent manner, and that this is associated with the increased expression of HAS2 in the presence of 20GPPD.

20GPPD induces ERK and Akt phosphorylation in HaCaT cells

It has previously been demonstrated that HAS2 expression is regulated by the MAPK and PI3K/Akt signaling pathways (23). Blocking p38 and MEK signaling has been shown to inhibit HAS2 expression by up to 90 and 40%, respectively, whereas the inhibition of JNK has no such effect (27). We therefore sought to determine the effects of 20GPPD on the phosphorylation of ERK and Akt. 20GPPD enhanced the phosphorylation of ERK and Akt in a dose-dependent manner, and these phosphorylation levels peaked at 0.5 and 1 h of 20GPPD treatment, respectively (Fig. 3). To examine whether these activated signaling pathways are associated with the 20GPPD-induced production of HA, pharmacological inhibitors of ERK and Akt (U0126 and LY294002, respectively) were employed. When the cells were treated with these inhibitors 1 h prior to treatment with 20GPPD, the 20GPPD-induced expression of HAS2 was diminished in both the U0126 and LY294002 treatment groups (Fig. 4A). Certain studies have reported that HA synthesis is mediated by the EGF receptor (EGFR) (15,28,29) and Ca2+-mediated signaling pathways, thus prompting us to investigate the effect of EGFR inhibition and intracellular Ca2+ inhibition on the 20GPPD-induced HAS2 expression. However, neither AG1478 (an EGFR inhibitor) nor BAPTA-AM (an intracellular Ca2+ chelator) affected the 20GPPD-induced expression of HAS2 (Fig. 4B). Overall, these results suggest that the 20GPPD-induced expression of HAS2 is mediated by the MEK and PI3K signaling pathways, rather than by EGFR or intracellular Ca2+ signaling in HaCaT cells.

Src regulates the 20GPPD-induced production of HA by inducing ERK and Akt phosphorylation

It has previously been demonstrated that Src regulates HAS2 expression in human dermal fibroblasts (23). To further determine whether 20GPPD induces Src signaling in human keratinocytes, Src phosphorylation was evaluated. Src phosphorylation was observed to be increased following treatment with 20GPPD in a dose-dependent manner at 20 min (Fig. 5A). Furthermore, the role of Src in the 20GPPD-induced production of HA was investigated using PP2, an Src inhibitor. When the cells were treated with PPT for 1 h prior to treatment with 20GPPD, the 20GPPD-increased production of HA (Fig. 5B) was observed to be decreased along with HAS2 expression (Fig. 5C). It has been reported that Src is an upstream regulator of ERK and Akt signaling. Zhang et al reported that Src mediates breast cancer cell migration by activating Akt and ERK1/2 (30), while the inhibition of Src in vascular smooth muscle cells has been observed to suppress ERK and Akt phosphorylation (31). We thus investigated the effects of Src inhibition by PP2 on the 20GPPD-induced phosphorylation of ERK and Akt in the HaCaT cells (Fig. 5D). 20GPPD significantly induced ERK and Akt phosphorylation, whereas treatment with PP2 abolished both the phosphorylation of ERK and Akt in a dose-dependent manner. These results suggest that Src mediates the effects of 20GPPD on the production of HA by activating ERK and Akt signaling.

Discussion

Increasing HA synthesis represents a promising strategy to improve skin hydration. HA is normally synthesized by one of 3 HAS enzymes; HAS1, HAS2 or HAS3. Tammi and Tammi (17), reported that HAS2 expression was reduced in the epidermis and dermis of intrinsically aged human skin. Therefore, the upregulation of HAS2 expression may represent a novel strategy for improving HA levels in the skin.

20GPPD is a major metabolite of ginsenoside Rb1, which has been shown to increase HAS2 mRNA expression and the HA content in human keratinocytes and increase epidermal thickness within the skin of a hairless mouse model (10). However, the mechanisms underlying these effects of 20GPPD remain unknown. In this study, we therefore examined the molecular mechanisms responsible for the enhancement of HA production by 20GPPD in HaCaT cells in an effort to verify its potential beneficial function in skin hydration. In a previous study, the bioavailability of 20GPPD was estimated using intravenous (i.v.) injection and oral administration in rats (32). The corresponding dose of 20GPPD (1 μM) was achieved in plasma concentration by both i.v. injection and oral administration. Furthermore, the authors demonstrated the rapid absorption of 20GPPD using Caco-2 cell permeability assay (32). Given that the highest dose of 20GPPD we used in this study was 1 μM, our results provide useful insight for the prediction of clinical outcomes.

Previous studies have demonstrated the upregulation of HAS through Akt and ERK (23). Our findings demonstrated that treatment with 20GPPD induced the phosphorylation of both ERK and Akt (Fig. 3A), but not JNK or p38 phosphorylation (unpublished data). Pharmacological inhibitors of ERK and Akt blocked the effects of 20GPPD on HAS2 expression, suggesting that ERK and Akt are the key players in 20GPPD-induced expression of HAS2. It has also been established that EGFR and Ca2+ signaling plays a critical role in keratinocyte homeostasis (3336). We thus employed AG1478 (an EGFR inhibitor) and BAPTA/AM (an intracellular Ca2+ chelator) to examine the involvement of EGFR Ca2+ signaling in the 20GPPD-increased expression of HAS2. However, neither AG1478 nor BAPTA/AM treatment affected the 20GPPD-induced expression of HAS. These results support our hypothesis that the 20GPPD-induced expression of HAS2 is mediated by ERK and Akt, but not by EGFR or a Ca2+-related signaling pathway.

Previous studies have indicated that Src mediates ERK and Akt signaling (37,38). In addition, Yang et al reported that Src and ERK enhance the HA oligosaccharide content in an endothelial cell model (39). In the present study, we demonstrated that Src mediated the 20GPPD-induced production of HA by activating ERK and Akt signaling. The increased HA content following treatment with 20GPPD was found to be abolished by PP2, an inhibitor of Src family kinase. In the group treated with 10 μM of PP2, the HA level was found to be lower than the level in the control group (Fig. 5B). This may be explained by the fact that PP2 is an inhibitor of several Src family kinases, rather than a specific Src inhibitor. As described above, studies have reported that Src is an upstream regulator of ERK and Akt (37,38). Indeed, we demonstrated that the 20GPPD-induced phosphorylation of ERK and Akt was decreased by the suppression of Src activity by PP2 (Fig. 5D). Therefore, we hypothesized that Src mediates ERK and Akt signaling during the 20GPPD-induced production of HA. It is notable that a previous study indicated that HAS2 can be regulated by O-linked N-acetylglucosamine (O-GlcNAcylation) on serine 221 (40). The effect of 20GPPD on this additional regulatory mechanism requires further investigation.

In conclusion, our data underline the potential skin hydrating effects of 20GPPD through the activation of HA production. Moreover, we found that Src tyrosine kinase is a key regulator of the 20GPPD-induced production of HA which occurs through the activation of ERK and Akt, but not through that of JNK or p38. To further support these findings, in vivo studies using an Src knockout model are warranted.

Acknowledgments

This study was supported by the National Leap Research Program (no. 2010-0029233) through the National Research Foundation funded by the Ministry of Science, ICT and Future Planning, Republic of Korea and by the R&D program of MOTIE/KIAT (Establishment of Infra Structure for Anti-aging Industry Support; No. N0000697), Republic of Korea.

References

1 

Christensen LP: Ginsenosides chemistry, biosynthesis, analysis, and potential health effects. Adv Food Nutr Res. 55:1–99. 2009. View Article : Google Scholar

2 

Kim WK, Song SY, Oh WK, et al: Wound-healing effect of ginsenoside Rd from leaves of Panax ginseng via cyclic AMP-dependent protein kinase pathway. Eur J Pharmacol. 702:285–293. 2013. View Article : Google Scholar : PubMed/NCBI

3 

Kimura Y, Sumiyoshi M, Kawahira K and Sakanaka M: Effects of ginseng saponins isolated from Red Ginseng roots on burn wound healing in mice. Br J Pharmacol. 148:860–870. 2006. View Article : Google Scholar : PubMed/NCBI

4 

Lee HS, Kim MR, Park Y, et al: Fermenting red ginseng enhances its safety and efficacy as a novel skin care anti-aging ingredient: in vitro and animal study. J Med Food. 15:1015–1023. 2012. View Article : Google Scholar : PubMed/NCBI

5 

He D, Sun J, Zhu X, Nian S and Liu J: Compound K increases type I procollagen level and decreases matrix metalloproteinase-1 activity and level in ultraviolet-A-irradiated fibroblasts. J Formos Med Assoc. 110:153–160. 2011. View Article : Google Scholar : PubMed/NCBI

6 

Lou JS, Chen XE, Zhang Y, et al: Photoprotective and immunoregulatory capacity of ginsenoside Rg1 in chronic ultraviolet B-irradiated BALB/c mouse skin. Exp Ther Med. 6:1022–1028. 2013.PubMed/NCBI

7 

Cai BX, Jin SL, Luo D, Lin XF and Gao J: Ginsenoside Rb1 suppresses ultraviolet radiation-induced apoptosis by inducing DNA repair. Biol Pharm Bull. 32:837–841. 2009. View Article : Google Scholar : PubMed/NCBI

8 

Cai BX, Luo D, Lin XF and Gao J: Compound K suppresses ultraviolet radiation-induced apoptosis by inducing DNA repair in human keratinocytes. Arch Pharm Res. 31:1483–1488. 2008. View Article : Google Scholar : PubMed/NCBI

9 

Cuong TT, Yang CS, Yuk JM, et al: Glucocorticoid receptor agonist compound K regulates Dectin-1-dependent inflammatory signaling through inhibition of reactive oxygen species. Life Sci. 85:625–633. 2009. View Article : Google Scholar : PubMed/NCBI

10 

Kim S, Kang BY, Cho SY, et al: Compound K induces expression of hyaluronan synthase 2 gene in transformed human keratinocytes and increases hyaluronan in hairless mouse skin. Biochem Biophys Res Commun. 316:348–355. 2004. View Article : Google Scholar : PubMed/NCBI

11 

Hwang JA, Hwang MK, Jang Y, et al: 20-O-β-d-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginseng, inhibits colon cancer growth by targeting TRPC channel-mediated calcium influx. J Nutr Biochem. 24:1096–1104. 2013. View Article : Google Scholar : PubMed/NCBI

12 

Laurent TC and Fraser JR: Hyaluronan. FASEB J. 6:2397–2404. 1992.PubMed/NCBI

13 

Oe M, Mitsugi K, Odanaka W, et al: Dietary hyaluronic acid migrates into the skin of rats. Scientific World Journal. 2014:3780242014. View Article : Google Scholar : PubMed/NCBI

14 

Toole BP: Hyaluronan in morphogenesis. J Intern Med. 242:35–40. 1997. View Article : Google Scholar : PubMed/NCBI

15 

Pasonen-Seppänen SM, Maytin EV, Törrönen KJ, et al: All-trans retinoic acid-induced hyaluronan production and hyperplasia are partly mediated by EGFR signaling in epidermal keratinocytes. J Invest Dermatol. 128:797–807. 2008. View Article : Google Scholar

16 

Stern R and Maibach HI: Hyaluronan in skin: aspects of aging and its pharmacologic modulation. Clin Dermatol. 26:106–122. 2008. View Article : Google Scholar : PubMed/NCBI

17 

Tammi RH and Tammi MI: Hyaluronan accumulation in wounded epidermis: a mediator of keratinocyte activation. J Invest Dermatol. 129:1858–1860. 2009. View Article : Google Scholar : PubMed/NCBI

18 

Weigel PH, Hascall VC and Tammi M: Hyaluronan synthases. J Biol Chem. 272:13997–14000. 1997. View Article : Google Scholar : PubMed/NCBI

19 

Spicer AP and McDonald JA: Characterization and molecular evolution of a vertebrate hyaluronan synthase gene family. J Biol Chem. 273:1923–1932. 1998. View Article : Google Scholar : PubMed/NCBI

20 

Camenisch TD, Spicer AP, Brehm-Gibson T, et al: Disruption of hyaluronan synthase-2 abrogates normal cardiac morphogenesis and hyaluronan-mediated transformation of epithelium to mesenchyme. J Clin Invest. 106:349–360. 2000. View Article : Google Scholar : PubMed/NCBI

21 

Tammi RH, Passi AG, Rilla K, et al: Transcriptional and post-translational regulation of hyaluronan synthesis. FEBS J. 278:1419–1428. 2011. View Article : Google Scholar : PubMed/NCBI

22 

Pasonen-Seppänen S, Karvinen S, Törrönen K, et al: EGF upregulates, whereas TGF-beta downregulates, the hyaluronan synthases Has2 and Has3 in organotypic keratinocyte cultures: correlations with epidermal proliferation and differentiation. J Invest Dermatol. 120:1038–1044. 2003. View Article : Google Scholar : PubMed/NCBI

23 

Li L, Asteriou T, Bernert B, Heldin CH and Heldin P: Growth factor regulation of hyaluronan synthesis and degradation in human dermal fibroblasts: importance of hyaluronan for the mitogenic response of PDGF-BB. Biochem J. 404:327–336. 2007. View Article : Google Scholar : PubMed/NCBI

24 

Thomas SM and Brugge JS: Cellular functions regulated by Src family kinases. Annu Rev Cell Dev Biol. 13:513–609. 1997. View Article : Google Scholar : PubMed/NCBI

25 

Bourguignon LY, Zhu H, Shao L and Chen YW: CD44 interaction with c-Src kinase promotes cortactin-mediated cytoskeleton function and hyaluronic acid-dependent ovarian tumor cell migration. J Biol Chem. 276:7327–7336. 2001. View Article : Google Scholar

26 

Schlessinger J: New roles for Src kinases in control of cell survival and angiogenesis. Cell. 100:293–296. 2000. View Article : Google Scholar : PubMed/NCBI

27 

Stuhlmeier KM and Pollaschek C: Differential effect of transforming growth factor beta (TGF-beta) on the genes encoding hyaluronan synthases and utilization of the p38 MAPK pathway in TGF-beta-induced hyaluronan synthase 1 activation. J Biol Chem. 279:8753–8760. 2004. View Article : Google Scholar

28 

Monslow J, Sato N, Mack JA and Maytin EV: Wounding-induced synthesis of hyaluronic acid in organotypic epidermal cultures requires the release of heparin-binding egf and activation of the EGFR. J Invest Dermatol. 129:2046–2058. 2009. View Article : Google Scholar : PubMed/NCBI

29 

Oh JH, Kim YK, Jung JY, Shin JE and Chung JH: Changes in glycosaminoglycans and related proteoglycans in intrinsically aged human skin in vivo. Exp Dermatol. 20:454–456. 2011. View Article : Google Scholar : PubMed/NCBI

30 

Zhang L, Teng Y, Zhang Y, et al: C-Src-mediated RANKL-induced breast cancer cell migration by activation of the ERK and Akt pathway. Oncol Lett. 3:395–400. 2012.PubMed/NCBI

31 

Liu G, Hitomi H, Hosomi N, et al: Prorenin induces vascular smooth muscle cell proliferation and hypertrophy via epidermal growth factor receptor-mediated extracellular signal-regulated kinase and Akt activation pathway. J Hypertens. 29:696–705. 2011. View Article : Google Scholar : PubMed/NCBI

32 

Paek IB, Moon Y, Kim J, et al: Pharmacokinetics of a ginseng saponin metabolite compound K in rats. Biopharm Drug Dispos. 27:39–45. 2006. View Article : Google Scholar

33 

Karvinen S, Pasonen-Seppänen S, Hyttinen JM, et al: Keratinocyte growth factor stimulates migration and hyaluronan synthesis in the epidermis by activation of keratinocyte hyaluronan synthases 2 and 3. J Biol Chem. 278:49495–49504. 2003. View Article : Google Scholar : PubMed/NCBI

34 

Kim do Y, Park MW, Yuan HD, Lee HJ, Kim SH and Chung SH: Compound K induces apoptosis via CAMK-IV/AMPK pathways in HT-29 colon cancer cells. J Agric Food Chem. 57:10573–10578. 2009. View Article : Google Scholar : PubMed/NCBI

35 

Lee JH, Jeong SM, Kim JH, et al: Effects of ginsenosides and their metabolites on voltage-dependent Ca(2+) channel subtypes. Mol Cells. 21:52–62. 2006.PubMed/NCBI

36 

Saavalainen K, Pasonen-Seppänen S, Dunlop TW, Tammi R, Tammi MI and Carlberg C: The human hyaluronan synthase 2 gene is a primary retinoic acid and epidermal growth factor responding gene. J Biol Chem. 280:14636–14644. 2005. View Article : Google Scholar : PubMed/NCBI

37 

Tang ZN, Zhang F, Tang P, Qi XW and Jiang J: RANKL-induced migration of MDA-MB-231 human breast cancer cells via Src and MAPK activation. Oncol Rep. 26:1243–1250. 2011.PubMed/NCBI

38 

Zhuang S, Duan M and Yan Y: Src family kinases regulate renal epithelial dedifferentiation through activation of EGFR/PI3K signaling. J Cell Physiol. 227:2138–2144. 2012. View Article : Google Scholar

39 

Yang CX, Liu YW, He YQ and Gao F: Src kinase-MAPK signal pathway plays a role in proliferation of endothelial cells induced by o-HA. Fen Zi Xi Bao Sheng Wu Xue Bao. 39:495–501. 2006.In Chinese.

40 

Vigetti D, Deleonibus S, Moretto P, et al: Role of UDP-N-acetylglucosamine (GlcNAc) and O-GlcNAcylation of hyaluronan synthase 2 in the control of chondroitin sulfate and hyaluronan synthesis. J Biol Chem. 287:35544–35555. 2012. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

May-2015
Volume 35 Issue 5

Print ISSN: 1107-3756
Online ISSN:1791-244X

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Lim T, Jeon AJ, Yoon JH, Song D, Kim J, Kwon JY, Kim JR, Kang NJ, Park J, Yeom MH, Yeom MH, et al: 20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginsenoside Rb1, enhances the production of hyaluronic acid through the activation of ERK and Akt mediated by Src tyrosin kinase in human keratinocytes. Int J Mol Med 35: 1388-1394, 2015
APA
Lim, T., Jeon, A.J., Yoon, J.H., Song, D., Kim, J., Kwon, J.Y. ... Lee, K.W. (2015). 20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginsenoside Rb1, enhances the production of hyaluronic acid through the activation of ERK and Akt mediated by Src tyrosin kinase in human keratinocytes. International Journal of Molecular Medicine, 35, 1388-1394. https://doi.org/10.3892/ijmm.2015.2121
MLA
Lim, T., Jeon, A. J., Yoon, J. H., Song, D., Kim, J., Kwon, J. Y., Kim, J. R., Kang, N. J., Park, J., Yeom, M. H., Oh, D., Lim, Y., Lee, C. C., Lee, C. Y., Lee, K. W."20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginsenoside Rb1, enhances the production of hyaluronic acid through the activation of ERK and Akt mediated by Src tyrosin kinase in human keratinocytes". International Journal of Molecular Medicine 35.5 (2015): 1388-1394.
Chicago
Lim, T., Jeon, A. J., Yoon, J. H., Song, D., Kim, J., Kwon, J. Y., Kim, J. R., Kang, N. J., Park, J., Yeom, M. H., Oh, D., Lim, Y., Lee, C. C., Lee, C. Y., Lee, K. W."20-O-β-D-glucopyranosyl-20(S)-protopanaxadiol, a metabolite of ginsenoside Rb1, enhances the production of hyaluronic acid through the activation of ERK and Akt mediated by Src tyrosin kinase in human keratinocytes". International Journal of Molecular Medicine 35, no. 5 (2015): 1388-1394. https://doi.org/10.3892/ijmm.2015.2121