Ethanolic Extract of Nerium indicum Mill. Decreases Transforming Growth Factor Beta-1 and Vascular Endothelial Growth Factor Expressions in Keloid Fibroblasts

Hernita  Taurustya; Mae Sri Hartati  Wahyuningsih; Indwiani Astuti

doi:10.3889/oamjms.2020.4292

Authors

Hernita Taurustya Department of Pharmacology and Therapy, Faculty of Medicine and Health Sciences, Universitas Bengkulu, Bengkulu, Indonesia
Mae Sri Hartati Wahyuningsih Department of Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia; Center for Herbal Medicine, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada. Yogyakarta, Indonesia
Indwiani Astuti Department of Pharmacology and Therapy, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia

DOI:

https://doi.org/10.3889/oamjms.2020.4292

Keywords:

Keloid fibroblast, Nerium indicum Mill, Oleandrin, TGF-Beta1, VEGF

Abstract

BACKGROUND: Keloid is a benign fibroproliferative dermis tumor characterized by an increase in growth factors which induce fibroblast proliferation, excessive migration, and synthesis of collagen. Nerium indicum Mill. extract had been studied as a keloid therapy agent. 5α-oleandrin contained in N. indicum has antikeloid activity by inhibiting keloid fibroblast proliferation, fibroblast migration, collagen deposition, and transforming growth factor beta-1 (TGF-β1) synthesis.

OBJECTIVE: This study aimed to determine the effect of administration of N. indicum extract on TGF-β1 and vascular endothelial growth factor (VEGF) expression in keloid fibroblast.

METHODS: This research was a quasi-experimental research with a post-test only control group design. The research subjects were fibroblast cells passage IV-VII isolated from patients’ keloid tissue with explant techniques. Treatment groups received N. indicum extract with a serial concentration of 2 μg/ml, 1 μg/ml, and 0.5 μg/ml, and control group received medium only. The supernatant was obtained after 72 h incubation period. Examination of TGF-β1 and VEGF expressions was performed using ELISA procedure.

RESULT: The expression of TGF-β1 in the treatment groups of the extract N. indicum (2 μg/ml, 1 μg/ml, and 0.5 μg/ml) was significantly lower than a control group of keloid fibroblasts (p < 0.05), according to increased concentration. VEGF expression in the treatment groups of N. indicum extract was lower compared to the control group of keloid fibroblasts. A significant decrease in keloid fibroblast VEGF levels occurred at extract concentrations of 2 μg/ml and 1 μg/ml (p < 0.05).

CONCLUSION: N. indicum extract could decrease TGF-β1 and VEGF expressions compared to control medium in keloid fibroblast cultures.

Downloads

Download data is not yet available.

Metrics

Metrics Loading ...

Plum Analytics Artifact Widget Block

References

Seo BF, Lee JY, Jung SN. Models of abnormal scarring. Biomed Res Int. 2013;2013:423147. PMid:24078916

Velnar T, Bailey T, Smrkolj V. The wound healing process: An overview of the cellular and molecular mechanisms. J Int Med Res. 2009;37(5):1528-42. https://doi. org/10.1177/147323000903700531 PMid:19930861

Shih B, Garside E, McGrouther DA, Bayat A. Molecular dissection of abnormal wound healing processes resulting in keloid disease. Wound Repair Regen. 2010;18(2):139-53. https://doi.org/10.1111/j.1524-475x.2009.00553.x PMid:20002895

Chau CH, Clavijo CA, Deng HT, Zhang Q, Kim KJ, Qiu Y, et al. Etk/bmx mediates expression of stress-induced adaptive genes VEGF, Pai-1, and iNOS via multiple signaling cascades in different cell systems. Am J Physiol Cell Physiol. 2005;289(2):C444-54. https://doi.org/10.1152/ajpcell.00410.2004 PMid:15788485

Slemp AE, Kirschner RE. Keloids and scars: A review of keloids and scars, their pathogenesis, risk factors, and management. Curr Opin Pediatr. 2006;18(4):396-402. https:// doi.org/10.1097/01.mop.0000236389.41462.ef PMid:16914994

Wahyuningsih MS, Mubarika S, Bolhuis R, Nooter K, Gandjar IG, Wahyuono S. Cytotoxicity of oleandrin isolated from the leaves of Nerium indicum mill. On several human cancer cell lines. Indones J Pharm. 2004;15(2):96-103.

Mae SH, Sofia M, Bolhuis RL, Nooter K, Oostrum RG, Subagus W, et al. Selectivity of compounds isolated from the leaves of Nerium indicum mill. On various human cancer cell lines. Med J Malaysia. 2008;63 Suppl A:24-5. PMid:19024965

Dachlan I, Wirohadidjojo YW, Wahyuningsih MS, Aryandono T, Soebono H, Afandy D. The effect of 5α-oleandrin on keloid fibroblast activities. BMC Proc. 2019;13 Suppl 11:14. https://doi. org/10.1186/s12919-019-0177-6 PMid:31890007

Wahyuningsih MS, Yuliani FS, Rahmawati DY, Pratiwi AN. Antifibrotic effect of ethanolic extract of Nerium indicum mill. Standardized 5α-oleandrin on keloid fibroblasts cells. Trad Med J. 2018;23(1):70-8. https://doi.org/10.22146/mot.35116

Tang M, Bian W, Cheng L, Zhang L, Jin R, Wang W, et al. Ginsenoside Rg3 inhibits keloid fibroblast proliferation, angiogenesis and collagen synthesis in vitro via the tgfβ/smad and erk signaling pathways. Int J Mol Med. 2018;41(3):1487-99. https://doi.org/10.3892/ijmm.2018.3362 PMid:29328420

Gonçalves-de-Albuquerque CF, Silva AR, da Silva CI, Castro-Faria-Neto HC, Burth P. Na/k pump and beyond: Na/k-atpase as a modulator of apoptosis and autophagy. Molecules. 2017;22(4):578. https://doi.org/10.3390/molecules22040578 PMid:28430151

Wang XQ, Lu SL, Mao ZG, Liu YK. Study on the biological function of vascular endothelial cells in the hypertrophic scar. Zhonghua Shao Shang Za Zhi. 2007;23(3):219-21. PMid:18019066

Bran GM, Goessler UR, Schardt C, Hormann K, Riedel F, Sadick H. Effect of the abrogation of tgf-β1 by antisense oligonucleotides on the expression of tgf-β-isoforms and their receptors I and II in isolated fibroblasts from keloid scars. Int J Mol Med. 2010;25(6):915-21. https://doi.org/10.3892/ ijmm_00000422 PMid:20428796

Manna SK, Sah NK, Newman RA, Cisneros A, Aggarwal BB. Oleandrin suppresses activation of nuclear transcription factor-κb, activator protein-1, and c-jun nh2-terminal kinase. Cancer Res. 2000;60(14):3838-47. PMid:10919658

Lee CH, Hong CH, Chen YT, Chen YC, 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. 2012;67(3):173-80. https://doi. org/10.1016/j.jdermsci.2012.06.004 PMid:22771320

Viera MH, Vivas AC, Berman B. Update on keloid management: Clinical and basic science advances. Adv Wound Care. 2012;1(5):200-6. https://doi.org/10.1089/ wound.2011.0313 PMid:24527306

Wahyuningsih MS, Nugrahaningsih DA, Budiyanto A. Ethanolic extract of Tithonia diversifolia (hemsley) a. Gray inhibits migration activity and degrease the transforming growth factor-beta1, VEGF expression on keloid fibroblasts. Asian J Pharm Clin Res. 2019;12(1):342-5. https://doi.org/10.22159/ ajpcr.2019.v12i1.29850

Wu WS, Wang FS, Yang KD, Huang CC, Kuo YR. Dexamethasone induction of keloid regression through effective suppression of vegf expression and keloid fibroblast proliferation. J Invest Dermatol. 2006;126(6):1264-71. https:// doi.org/10.1038/sj.jid.5700274 PMid:16575391

Olsson AK, Dimberg A, Kreuger J, Claesson-Welsh L. Vegf receptor signalling? In control of vascular function. Nat Rev Mol Cell Biol. 2006;7(5):359. https://doi.org/10.1038/nrm1911 PMid:16633338

Karamysheva AF. Mechanisms of angiogenesis. Biochemistry (Mosc). 2008;73(7):751. PMid:18707583

Hicklin DJ, Ellis LM. Role of the vascular endothelial growth factor pathway in tumor growth and angiogenesis. J Clin Oncol. 2005;23(5):1011-27. https://doi.org/10.1200/jco.2005.06.081 PMid:15585754

Fearnley G, Smith G, Harrison M, Wheatcroft S, Tomlinson D, Ponnambalam SJ. Vascular endothelial growth factor-a regulation of blood vessel sprouting in health and disease. OA Biochemistry 2013;1(1):1. https://doi.org/10.13172/2052-9651-1-1-471