Sains Malaysiana 47(10)(2018): 2401–2409

http://dx.doi.org/10.17576/jsm-2018-4710-16

 

Kesan Polifenol Minyak Kelapa Dara ke atas Aktiviti Enzim Antioksidan dan Lipid Peroksida Pada Sel Osteoblas

(Virgin Coconut Oil Polyphenol Effects on Antioxidant Enzyme Activities and Lipid Peroxidation on Osteoblast Cell)

 

ZIL HAYATULLINA, NORAZLINA MOHAMED, IMA-NIRWANA SOELAIMAN & NORLIZA MUHAMMAD*

 

Jabatan Farmakologi, Fakulti Perubatan, Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Wilayah Persekutuan, Malaysia

 

Diserahkan: 3 April 2018/Diterima: 5 Jun 2018

 

ABSTRAK

Osteoporosis dikaitkan dengan radikal bebas dan tekanan oksidatif. Kandungan polifenol yang tinggi dalam minyak kelapa dara (VCO) yang bertindak sebagai antioksidan mampu menghalang tekanan oksidatif seterusnya mencegah osteoporosis. Uji kaji in vitro ini dijalankan untuk mengkaji mekanisme kesan perlindungan polifenol ekstrak daripada VCO (PF) ke atas sel selanjar pre-osteoblas mencit (MC3T3-E1) teraruh tekanan oksidatif. Untuk mengkaji viabiliti kesan perlindungan polifenol terhadap MC3T3-E1 daripada toksisiti hidrogen peroksida (H2O2), sel MC3T3-E1 dirawat dengan beberapa dos polifenol selama 24 jam selepas dieram dengan 250 μM (IC50) H2O2 selama 1 jam. Mekanisme perlindungan dikaji dengan mengukur aras malondialdehid (MDA) serta aktiviti enzim antioksidan iaitu superoksida dismutase (SOD), glutation peroksidase (GPx) dan katalase (CAT). Keputusan kajian menunjukkan bahawa kepekatan perencatan median (IC50) H2O2 pada 1 jam adalah 250 μM. Ujian viabiliti sel mendapati kesemua dos polifenol dapat melindungi sel MC3T3-E1 daripada toksisiti H2O2. Dos 1 dan 0.01 μM polifenol dapat meningkatkan aras GPx dan CAT, manakala dos 0.01 dan 0.0001 μM dapat meningkatkan aras SOD. Sementara itu, hanya dos 0.01 dan 0.001 μM sahaja yang dapat menghalang peningkatan aras MDA. Kesimpulan yang dapat dibuat di dalam kajian ini adalah polifenol daripada minyak kelapa dara mencegah lipid peroksida pada sel MC3T3-E1 yang telah diaruh dengan H2O2 dengan meningkatkan aktiviti enzim antioksidan.

 

Kata kunci: Osteoporosis; polifenol; sel MC3T3-E1; tekanan oksidatif; VCO

 

ABSTRACT

Osteoporosis is associated with free radicals and oxidative stress. The high polyphenol contents in virgin coconut oil (VCO) can prevent oxidative stress and consequently osteoporosis. This in vitro study was conducted in order to determine the mechanism of the protective effects of polyphenol extracted from VCO on murine osteoblast-like cell line (MC3T3-E1) which was induced with oxidative stress. To study the protective effects on viability of MC3T3-E1 cells exposed to hydrogen peroxide (H2O2) toxicity, MC3T3-E1 cells were post-treated with different doses of polyphenol for 24 h after 1 h incubation with 250 μM of H2O2 (IC50). The protective mechanism was studied by measuring melondialdehyde (MDA) levels as well as antioxidant enzymes activities which are superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase (CAT). The results showed that the median inhibitary concentration (IC50) for H2O2 after 1 h was 250 μM. Cell viability tests showed that all the doses of polyphenol were able to protect MC3T3-E1 cells from H2O2 toxicity. The dose of 1 and 0.01 μM of polyphenol increased the level of GPx and CAT while the dose of 0.01 and 0.0001 μM increased the level of SOD. Meanwhile, the 0.0001 μM of PF prevented the MDA elevation. In conclucion, the polyphenol components in virgin coconut oil prevent lipid peroxidation in MC3T3-E1 cells induced with H2O2 by increasing the endogenous antioxidant enzymes.

 

Keywords: MC3T3-E1 cells; osteoporosis; oxidative stress; polyphenol; VCO

RUJUKAN

Abujazia, M.A., Norliza, M., Ahmad, N.S. & Ima Nirwana, S. 2012. The effects of virgin coconut oil on bone oxidative status in ovariectomized rats. Evidence-Based Complementary & Alternative Medicine 2012: 525079. http:// dx.doi.org/10.1155/2012/525079.

Altindag, O., Erel, O., Soran, N., Celik, H. & Selek, S. 2008. Total oxidative/antioxidative status and relation to bone mineral density in osteoporosis. Rheumatology International 28(4): 317-321.

Arora, R., Chawla, R., Marwah, R., Arora, P., Sharma, R.K., Kaushik, V., Goel, R., Kaur, A., Silambarasan, M., Tripathi, R.P. & J.R. Bhardwaj. 2011. Potential of complementary and alternative medicine in preventive management of novel H1N1 flu (Swine flu) pandemic: Thwarting potential disasters in the Bud. Evidence-Based Complementary & Alternative Medicine. 2011: 586506. doi:10.1155/2011/586506.

Baek, K.H., Oh, K.W., Lee, W.Y., Lee, S.S., Kim, M.K., Kwon, H.S., Rhee, E.J., Han J.H., Song, K.H. & Cha, B.Y. 2010. Association of oxidative stress with postmenopausal osteoporosis and the effects of hydrogen peroxide on osteoclast formation in human bone marrow cell cultures. Calcified Tissue International 87(3): 226-235.

Barnabé, W., de Mendonça Neto, T., Pimenta, F.C., Pegoraro, L.F. & Scolaro, J.M. 2004. Efficacy of sodium hypochlorite and coconut soap used as disinfecting agents in the reduction of denture stomatitis, Streptococcus mutans and Candida albicans. Journal of Oral Rehabilitation 31(5): 453-459.

Chodimella, R., Anderson, J., Hong, Y. & Bishop, A. 2005. Nitric Oxide (NO) Sensitivity and Differentiation Status in Motor Neurons (Abstract). Washington, DC: Society for Neuroscience.

Cosman, F., de Beur, S.J., LeBoff, M.S., Lewiecki, E.M., Tanner, B., Randall, S. & Lindsay, R. 2014. Clinician’s guide to prevention and treatment of osteoporosis. Osteoporosis International 25(10): 2359-2381.

Danielle, A.C. & Jiang, J.X. 2015. Reactive oxygen species and oxidative stress in osteoclastogenesis, skeletal aging and bone diseases. Journal of Bone and Mineral Metabolism 33(4): 359-370.

Daroszewska, A. 2015. Prevention and treatment of osteoporosis in women: An update. Obstetrics, Gynaecology & Reproductive Medicine 25(7): 181-187.

Denisova, N.A., Cantuti-Castelvetri, I., Hassan, W.N., Paulson, K.E. & Joseph, J.A. 2001. Role of membrane lipids in regulation of vulnerability to oxidative stress in PC12 cells: implication for aging. Free Radical Biology and Medicine 30(6): 671-678.

Dosumu, O.O., Duru, F.I.O., Osinuba, A.A., Oremosu, A.A. & Noronha, C.C. 2010. Influence of virgin coconut oil (VCNO) on oxidative stress, serum testosteron and gonadotropic hormones (FSH, LH) in chronic ethanol ingestion. Agriculture & Biology Journal of North America 1(6): 1126-1132.

Fatokun, A.A., Stone, T.W. & Smith, R.A. 2008. Responses of differentiated MC3T3-E1 osteoblastic cells to reactive oxygen species. European Journal of Pharmacology 587(1- 3): 35-41.

Hagiwara, K., Goto, T., Araki, M., Miyazaki, H. & Hagiwara, H. 2011. Olive polyphenol hydroxytyrosol prevents bone loss. European Journal of Pharmacology 662(1-3): 78-84.

Kamat, J.P., Boloor, K.K., Devasagayam, T.P.A. & Venkatachalam, S.R. 2000. Antioxidant properties of asparagus racemosus against damage induced by gamma-radiation in rat liver mitochondria. Journal of Ethnopharmacology 71(3): 425- 435.

Lee, Y.S., Chen, X. & Anderson, J.J.B. 2001. Physiological concentrations of genistein stimulate the proliferation and protect against free radical-induced oxidative damage of MC3T3-E1 osteoblast-like cells. Nutrition Research 21(9): 1287-1298.

Linares, G.R., Xing, W., Govoni, K.E., Chen, S. & Mohan, S. 2009. Glutaredoxin 5 regulates osteoblast apoptosis ny protecting against oxidative stress. Bone 44(5): 795-804.

Maggio, D., Barabani, M., Pierandrei, M., Polidori, M.C., Catani, M., Mecocci, P., Senin, U., Pacifici, R. & Cherubini, A. 2003. Marked decrease in plasma antioxidants in aged osteoporotic women: results of a cross-sectional study. Journal of Clinical Endocrinology & Metabolism 88(4): 1523-1527.

Manisha, D. & Shyamapada, M. 2011. Coconut (Cocos nucifera L.: Arecaceae): In health promotion and disease prevention. Asian Pacific Journal of Tropical Medicine 4(3): 241-247.

Marina, A.M., Che Man, Y.B. & Nazimah, A.H. 2009. Chemical properties of virgin coconut oil. Journal of the American Oil Chemists’ Society 86(4): 301-307.

McNamara, L.M. 2010. Perpective on post-menopausal osteoporosis: Establishing an interdisciplinary understanding of the sequence of events from the molecular level to whole bone fracktures. Journal of the Royal Society Interface 7(44): 353-372.

Morton, D.J., Barrett-Connor, E.L. & Schneider, D.L. 2001. Vitamin C supplement use and bone mineral density in postmenopausal women. Journal of Bone Mineral Research 16(1): 135-140.

Muhammad, Z.M., Ahmad, N.S., Norazlina, M., Norliza, M. & Ima-Nirwana, S. 2010. Beneficial effects of vitamin E isomer supplementation on static and dynamic bone histomorphometry parameters in normal male rats. Journal of Bone and Mineral Metabolism 28(5): 503-509.

Nadia, M.E. & Ahmad, N.S. 2014. Time and dose-dependent effects of Labisia pumila on bone oxidative status of postmenopausal osteoporosis rat model. Nutrients 6(8): 3288-3302.

Nevin, K.G. & Rajamohan, T. 2006. Virgin coconut oil supplemented diet increased the antioxidant status in rats. Food Chemistry 99(2): 260-266.

Nevin, K.G. & Rajamohan, T. 2008. Influence of virgin coconut oil on blood coagulation factors, lipid levels and LDL oxidation in cholesterol fed Sprague-Dawley rats. European Journal of Clinical Nutrition Metabolism 3: 1-8.

Nevin, K.G. & Rajamohan, T. 2005. Virgin coconut oil supplemented diet increases the antioxidant status in rats. Food Chemistry 99(2): 260-266.

Nevin, K.G. & Rajamohan, T. 2004. Beneficial effects of virgin coconut oil on lipid parameters and in vitro LDL oxidation. Clinical Biochemistry 37(9): 830-835.

Nizar, A.M., Norazlina, M. & Ahmad, N.S. 2012. Effects of low-dose versus high doses γ-tocotrienol on the bone cells exposed to the hydrogen peroxide-induced oxidative stress and apoptosis. Evidence-Based Complementary & Alternative Medicine 2012: 680834. http://dx.doi. org/10.1155/2012/680834.

Norliza, M., Douglas, A.L., Ahmad, N.S., Norazlina, M. & Ima-Nirwana, S. 2012. Two different isomers of vitamin E prevent bone loss in postmenopausal osteoporosis rat model. Evidence-Based Complementary & Alternative Medicine 2012: 161527. https://doi.org/10.1155/2012/161527.

Oyi, A.R., Onaolapo, J.A. & Obi, R.C. 2010. Formulation and antimicrobial studies of coconut (Cocos nucifera Linne) oil. Research Journal of Applied Sciences, Engineering and Technology 2(2): 133-137.

Park, B.G., Yoo, C.I., Kim, H.T., Kwon, C.H. & Kim, Y.K. 2005. Role of mitogen- activated protein kinases in hydrogen peroxide-induced cell death in osteoblastic cells. Toxicology 215(1-2): 115-125.

Park, Y.H., Wan, D.W., Suh, H., Ryu, G.H., Hyon, S.H., Cho, B.K. & Park, J.C. 2003. Protective effects of green tea polyphenol against reactive oxygen species-induced oxidative stress in cultures rat calvarial osteoblast. Cell Biology & Toxicology 19(5): 325-337.

Pereira, R.M., Delany, A.M. & Canalis, E. 2001. Cortisol inhibits the differentiation and apoptosis of osteoblasts in culture. Bone 28(5): 484-490.

Rupesh, D., Chitrangada, A., Bindu, P.C. & Kundu, S.C. 2008. Antioxidant potential of silk protein sericin against hydrogen peroxide-induced oxidative stress in skin fibroblasts. BMP Reports 41(3): 236-241.

Satoh, I., Sakai, N., Enokido, Y., Uchiyama, Y. & Hatanaka, H. 1996. Free radical independent protection by nerve growth factor and Bcl-2 of PC12 cells from hydrogen peroxide-triggered apoptosis. Journal of Biochemistry 120: 540-546.

Scalbert, A., Johnson, I.T. & Saltmarsh, M. 2005. Polyphenols: Antioxidants and beyond. American Journal of Clinical Nutrition 81(1): 215S-217S.

Sendur, O.F., Turan, Y., Tastaban, E. & Serter, M. 2009. Antioxidant status in patients with osteoporosis: A controlled study. Joint Bone Spine 76(5): 514-518.

Sosnowska, J. & Balslev, H. 2009. American palm ethnomedicine: A metaanalysis. Journal of Ethnobiology and Ethnomedicine 5: 43. https://doi.org/10.1186/1746-4269-5-43.

Taheri, J.B., Espineli, F.W., Lu, H., Asayesh, M., Bakshi, M. & Nakhostin, M.R. 2010. Antimicrobial effect of coconut flour on oral microflora: An in vitro study. Research Journal of Biological Sciences 5(6): 456-459.

Wang, Y.K., Hong, Y.J., Wei, M., Wu, Y., Huang, Z.Q., Chen, R.Z. & Chen, H.Z. 2010. Curculigoside attenuates human umbilicak vein endothelial cell injury induced by H2O2. Journal of Ethnopharmacology 132(1): 233-239.

Wu, J.H., Xu, C., Shan, C.Y. & Tan, R.X. 2006. Antioxidant properties and PC12 cell protective effects of APS-1, a polysaccharide from Aloe vera var. Chinensis. Life Science 78(6): 622-630.

Xu, Z.S., Wang, X.Y., Xiao, D.M., Hu, L.F., Lu, M., Wu, Z.Y. & Bian, J.S. 2011. Hydrogen sulfide protects MC3T3-E1 osteoblastic cells against H2O2-induced oxidative damage - implications for the treatmet of osteoporosis. Free Radical Biology & Medicine 50(10): 1314-1323.

Yin, H., Xu, L. & Porter, N.A. 2011. Free radical lipid peroxidation: Mechanisms and analysis. Chemical Reviews 111(10): 5944-5972.

Zalavras, C., Shah, S., Birnbaum, M.J. & Frenkel, B. 2003. Role of apoptosis in glucocorticoid-induced osteoporosis and osteonecrosis. Critical Reviews in Eukaryotic Gene Expression 13(2-4): 221-235.

Zhang, J.K., Yang, L., Meng, G.L., Fang, J., Chen, J.Z., He, Q.Z., Chen, S., Fan, J.Z., Luo, Z.J. & Liu, J. 2012. Protective effects of tetrehydroxystilbene glucoside against hydrogen peroxide-induced dysfunction and oxidative stress in osteoblastic MC3T3-E1 cell. European Journal of Pharmacology 689: 31-37.

Zil Hayatullina, Norliza, M., Norazlina, M. & Ima-Nirwana, S. 2012. Virgin coconut oil supplementation prevents bone loss in osteoporosis rat model. Evidence-Based Complementary and Alternative Medicine 2012: 237236.

 

*Pengarang untuk surat-menyurat; email: norliza_ssp@ppukm.ukm.edu.my

 

 

 

 

 

 

 

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