Ginsenoside Rg3 inhibits keloid fibroblast proliferation, angiogenesis and collagen synthesis in vitro via the TGF‑β/Smad and ERK signaling pathways

Tang,Mengyao; Bian,Weiwei; Cheng,Liying; Zhang,Lu; Jin,Rong; Wang,Wenbo; Zhang,Yuguang

doi:10.3892/ijmm.2018.3362

Ginsenoside Rg3 inhibits keloid fibroblast proliferation, angiogenesis and collagen synthesis in vitro via the TGF‑β/Smad and ERK signaling pathways

Authors:
- Mengyao Tang
- Weiwei Bian
- Liying Cheng
- Lu Zhang
- Rong Jin
- Wenbo Wang
- Yuguang Zhang
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Affiliations: Department of Plastic and Reconstructive Surgery, Shanghai Ninth People's Hospital Affiliated Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China
Published online on: January 4, 2018 https://doi.org/10.3892/ijmm.2018.3362
Pages: 1487-1499
Copyright: © Tang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A wide range of therapeutic options exists for the treatment of keloids, all of which have their own strengths; however, a high risk of side‑effects and frequent recurrence remains. Therefore, the present study aimed to identify improved therapeutic approaches or drugs for the treatment of keloids. Ginsenoside Rg3 (Rg3) has been reported to exert numerous antitumor effects, thus indicating that Rg3 may be a potential therapeutic agent that targets keloids. The present study determined the effects of Rg3 on human keloid fibroblasts (KFs) in vitro, and further explored the associated molecular and cellular mechanisms. Keloid scar specimens were obtained from patients, aged between 22 and 35 years, without systemic diseases and primary cells were isolated from keloid tissues. In each assay, KFs were divided into three groups and were cultured in medium with or without various concentrations of Rg3 (50 or 100 µg/ml). Cell viability assay, flow cytometry, quantitative polymerase chain reaction, cell migration assay, immunofluorescence staining, western blot analysis, Transwell cell invasion assay and immunohistochemical analysis were used to analyze the KFs and keloid explant cultures. The results of the present study demonstrated that Rg3 was able to exert an inhibitory effect on the transforming growth factor‑β/Smad and extracellular signal‑regulated kinase signaling pathways in KFs. The proliferation, migration, invasion, angiogenesis and collagen synthesis of KFs were markedly suppressed following treatment with Rg3. Furthermore, the results of an ex vivo assay indicated that Rg3 inhibited angiogenesis and reduced collagen accumulation in keloids. Significant statistical differences existed between the control and Rg3‑treated groups (P<0.05). All of these experimental results suggested that Rg3 may serve as a reliable drug for the treatment of patients with keloids.

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1	Trace AP, Enos CW, Mantel A and Harvey VM: Keloids and Hypertrophic Scars: A Spectrum of Clinical Challenges. Am J Clin Dermatol. 17:201–223. 2016. View Article : Google Scholar : PubMed/NCBI
2	English RS and Shenefelt Pd: Keloids and hypertrophic scars. Dermatol Surg. 25:631–638. 1999. View Article : Google Scholar : PubMed/NCBI
3	Wang R, Chen J, Zhang Z and Cen Y: Role of chymase in the local renin-angiotensin system in keloids: Inhibition of chymase may be an effective therapeutic approach to treat keloids. Drug Des Devel Ther. 9:4979–4988. 2015.PubMed/NCBI
4	Lu WS, Zheng XD, Yao XH and Zhang LF: Clinical and epidemiological analysis of keloids in Chinese patients. Arch Dermatol Res. 307:109–114. 2015. View Article : Google Scholar
5	Butler PD, Longaker MT and Yang GP: Current progress in keloid research and treatment. J Am Coll Surg. 206:731–741. 2008. View Article : Google Scholar : PubMed/NCBI
6	Bijlard E, Steltenpool S and Niessen FB: Intralesional 5-fluorouracil in keloid treatment: A systematic review. Acta Derm Venereol. 95:778–782. 2015.PubMed/NCBI
7	Ud-Din S and Bayat A: Strategic management of keloid disease in ethnic skin: A structured approach supported by the emerging literature. Br J Dermatol. 169(Suppl 3): 71–81. 2013. View Article : Google Scholar : PubMed/NCBI
8	Liu T, Peng YF, Jia C, Yang BH, Tao X, Li J and Fang X: Ginsenoside Rg3 improves erectile function in streptozotocin-induced diabetic rats. J Sex Med. 12:611–620. 2015. View Article : Google Scholar
9	Yuan HD, Quan HY, Zhang Y, Kim SH and Chung SH: 20(S)-Ginsenoside Rg3-induced apoptosis in HT-29 colon cancer cells is associated with AMPK signaling pathway. Mol Med Rep. 3:825–831. 2010.
10	He BC, Gao JL, Luo X, Luo J, Shen J, Wang L, Zhou Q, Wang YT, Luu HH, Haydon RC, et al: Ginsenoside Rg3 inhibits colorectal tumor growth through the downregulation of Wnt/β-catenin signaling. Int J Oncol. 38:437–445. 2011. View Article : Google Scholar
11	Wang JH, Nao JF, Zhang M and He P: 20(s)-ginsenoside Rg3 promotes apoptosis in human ovarian cancer HO-8910 cells through PI3K/Akt and XIAP pathways. Tumour Biol. 35:11985–11994. 2014. View Article : Google Scholar : PubMed/NCBI
12	Joo SS, Yoo YM, Ahn BW, Nam SY, Kim YB, Hwang KW and Lee DI: Prevention of inflammation-mediated neurotoxicity by Rg3 and its role in microglial activation. Biol Pharm Bull. 31:1392–1396. 2008. View Article : Google Scholar : PubMed/NCBI
13	Chen QJ, Zhang MZ and Wang LX: Gensenoside Rg3 inhibits hypoxia-induced VEGF expression in human cancer cells. Cell Physiol Biochem. 26:849–858. 2010. View Article : Google Scholar
14	Pazyar N, Omidian M and Jamshydian N: Ginseng as a potential novel addition to the antikeloid weaponry. Phytother Res. 26:1579–1580. 2012.PubMed/NCBI
15	Liu JP, Lu D, Nicholson RC, Zhao WJ, Li PY and Wang F: Toxicity of a novel anti-tumor agent 20(S)-ginsenoside Rg3: A 26-week intramuscular repeated administration study in rats. Food Chem Toxicol. 50:3388–3396. 2012. View Article : Google Scholar : PubMed/NCBI
16	Kim SM, Lee SY, Cho JS, Son SM, Choi SS, Yun YP, Yoo HS, Yoon DY, Oh KW, Han SB, et al: Combination of ginsenoside Rg3 with docetaxel enhances the susceptibility of prostate cancer cells via inhibition of NF-kappaB. Eur J Pharmacol. 631:1–9. 2010. View Article : Google Scholar : PubMed/NCBI
17	Livak and Schmittgen: Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar
18	Lan CC, Liu IH, Fang AH, Wen CH and Wu CS: Hyperglycaemic conditions decrease cultured keratinocyte mobility: Implications for impaired wound healing in patients with diabetes. Br J Dermatol. 159:1103–1115. 2008.PubMed/NCBI
19	Shin JU, Lee WJ, Tran TN, Jung I and Lee JH: Hsp70 knockdown by siRNA decreased collagen production in keloid fibroblasts. Yonsei Med J. 56:1619–1626. 2015. View Article : Google Scholar : PubMed/NCBI
20	Gill SE and Parks WC: Metalloproteinases and their inhibitors: Regulators of wound healing. Int J Biochem Cell Biol. 40:1334–1347. 2008. View Article : Google Scholar
21	Liang CJ, Yen YH, Hung LY, Wang SH, Pu CM, Chien HF, Tsai JS, Lee CW, Yen FL and Chen YL: Thalidomide inhibits fibronectin production in TGF-β1-treated normal and keloid fibroblasts via inhibition of the p38/Smad3 pathway. Biochem Pharmacol. 85:1594–1602. 2013. View Article : Google Scholar : PubMed/NCBI
22	Schäfer M and Werner S: Cancer as an overhealing wound: An old hypothesis revisited. Nat Rev Mol Cell Biol. 9:628–638. 2008. View Article : Google Scholar : PubMed/NCBI
23	Shin YM, Jung HJ, Choi WY and Lim CJ: Antioxidative, anti-inflammatory, and matrix metalloproteinase inhibitory activities of 20(S)-ginsenoside Rg3 in cultured mammalian cell lines. Mol Biol Rep. 40:269–279. 2013. View Article : Google Scholar
24	Dong X, Mao S and Wen H: Upregulation of proinflammatory genes in skin lesions may be the cause of keloid formation (Review). Biomed Rep. 1:833–836. 2013. View Article : Google Scholar
25	Lee WJ, Ahn HM, Roh H, Na Y, Choi IK, Lee JH, Kim Yo, Lew DH and Yun CO: Decorin-expressing adenovirus decreases collagen synthesis and upregulates MMP expression in keloid fibroblasts and keloid spheroids. Exp Dermatol. 24:591–597. 2015. View Article : Google Scholar : PubMed/NCBI
26	Rao KB, Malathi N, Narashiman S and Rajan ST: Evaluation of myofibroblasts by expression of alpha smooth muscle actin: A marker in fibrosis, dysplasia and carcinoma. J Clin Diagn Res. 8:ZC14–ZC17. 2014.
27	Chipev CC, Simman R, Hatch G, Katz AE, Siegel DM and Simon M: Myofibroblast phenotype and apoptosis in keloid and palmar fibroblasts in vitro. Cell death differ. 7:166–176. 2000. View Article : Google Scholar : PubMed/NCBI
28	Mun JH, Kim YM, Kim BS, Kim JH, Kim MB and Ko HC: Simvastatin inhibits transforming growth factor-β1-induced expression of type I collagen, CTGF, and α-SMA in keloid fibroblasts. Wound Repair Regen. 22:125–133. 2014. View Article : Google Scholar : PubMed/NCBI
29	Zhu R, Yue B, Yang Q, Ma Y, Huang G, Guan M, Avram MM and Lu Z: The effect of 595 nm pulsed dye laser on connective tissue growth factor (CTGF) expression in cultured keloid fibroblasts. Lasers Surg Med. 47:203–209. 2015. View Article : Google Scholar : PubMed/NCBI
30	Khoo YT, Ong CT, Mukhopadhyay A, Han HC, Do DV, Lim IJ and Phan TT: Upregulation of secretory connective tissue growth factor (CTGF) in keratinocyte-fibroblast coculture contributes to keloid pathogenesis. J Cell Physiol. 208:336–343. 2006. View Article : Google Scholar : PubMed/NCBI
31	Duncan MR and Berman B: Differential regulation of glycosaminoglycan, fibronectin, and collagenase production in cultured human dermal fibroblasts by interferon-alpha, -beta, and -gamma. Arch Dermatol Res. 281:11–18. 1989. View Article : Google Scholar : PubMed/NCBI
32	Wu Y, Peng Y, Gao D, Feng C, Yuan X, Li H, Wang Y, Yang L, Huang S and Fu X: Mesenchymal stem cells suppress fibroblast proliferation and reduce skin fibrosis through a TGF-β3-dependent activation. Int J Low Extrem Wounds. 14:50–62. 2015. View Article : Google Scholar : PubMed/NCBI
33	Yoshimoto H, Ishihara H, Ohtsuru A, Akino K, Murakami R, Kuroda H, Namba H, Ito M, Fujii T and Yamashita S: Overexpression of insulin-like growth factor-1 (IGF-I) receptor and the invasiveness of cultured keloid fibroblasts. Am J Pathol. 154:883–889. 1999. View Article : Google Scholar : PubMed/NCBI
34	Ma H, Cai H, Zhang Y, Wu J, Liu X, Zuo J, Jiang W, Ji G, Zhang Y, Liu C, et al: Membrane palmitoylated protein 3 promotes hepatocellular carcinoma cell migration and invasion via upregulating matrix metalloproteinase 1. Cancer Lett. 344:74–81. 2014. View Article : Google Scholar : PubMed/NCBI
35	Fujiwara M, Muragaki Y and Ooshima A: Keloid-derived fibroblasts show increased secretion of factors involved in collagen turnover and depend on matrix metalloproteinase for migration. Br J Dermatol. 153:295–300. 2005. View Article : Google Scholar : PubMed/NCBI
36	Uchida G, Yoshimura K, Kitano Y, Okazaki M and Harii K: Tretinoin reverses upregulation of matrix metalloproteinase-13 in human keloid-derived fibroblasts. Exp dermatol. 12(Suppl 2): 35–42. 2003. View Article : Google Scholar
37	Lee CH, Hong CH, Chen YT, Chen YC and Shen MR: TGF-beta1 increases cell rigidity by enhancing expression of smooth muscle actin: Keloid-derived fibroblasts as a model for cellular mechanics. J Dermatol Sci. 67:173–180. 2012. View Article : Google Scholar : PubMed/NCBI
38	Song R, Li G and Li S: Aspidin PB, a novel natural anti-fibrotic compound, inhibited fibrogenesis in TGF-β1-stimulated keloid fibroblasts via PI-3K/Akt and Smad signaling pathways. Chem Biol Interact. 238:66–73. 2015. View Article : Google Scholar : PubMed/NCBI
39	Branton MH and Kopp JB: TGF-β and fibrosis. Microbes Infect. 1:1349–1365. 1999. View Article : Google Scholar : PubMed/NCBI
40	Nakao A, Afrakhte M, Morén A, Nakayama T, Christian JL, Heuchel R, Itoh S, Kawabata M, Heldin NE, Heldin CH, et al: Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling. Nature. 389:631–635. 1997. View Article : Google Scholar : PubMed/NCBI
41	Fujiwara M, Muragaki Y and Ooshima A: Upregulation of transforming growth factor-beta1 and vascular endothelial growth factor in cultured keloid fibroblasts: relevance to angiogenic activity. Arch Dermatol Res. 297:161–169. 2005. View Article : Google Scholar : PubMed/NCBI
42	Wu Y, Zhang Q, Ann DK, Akhondzadeh A, Duong HS, Messadi DV and Le AD: Increased vascular endothelial growth factor may account for elevated level of plasminogen activator inhibitor-1 via activating eRK1/2 in keloid fibroblasts. Am J Physiol Cell Physiol. 286:C905–C912. 2004. View Article : Google Scholar
43	He S, Yang Y, Liu X, Huang W and Zhang X, Yang S and Zhang X: Compound Astragalus and Salvia miltiorrhiza extract inhibits cell proliferation, invasion and collagen synthesis in keloid fibroblasts by mediating transforming growth factor-β/Smad pathway. Br J Dermatol. 166:564–574. 2012. View Article : Google Scholar
44	Tuan TL, Wu H, Huang EY, Chong SS, Laug W, Messadi D, Kelly P and Le A: Increased plasminogen activator inhibitor-1 in keloid fibroblasts may account for their elevated collagen accumulation in fibrin gel cultures. Am J Pathol. 162:1579–1589. 2003. View Article : Google Scholar : PubMed/NCBI