SYBR-Green I fluorescence-based qRT-PCR of clove oil's anticancer, chemical characterization and antidiabetic activities
Main Article Content
Keywords
Syzygium aromaticum, Myrtaceate, Diabetes, Human bone cancer, GC-MS, Cas3 gene expression, α-amylase and β-actin
Abstract
Syzygium aromaticum is a valuable herbal plant in China, Europe, and Asia. It has been described as a powerful analgesic, anti-inflammatory, antioxidant, antifungal, and antibacterial agent. The goal of this work is to determine the phytoconstituents in clove oil derived from Syzygium aromaticum, a Myrtaceae family member. The GC-MS analysis of clove essential oil revealed 23 chemical compounds, including eugenol (58.65%), caryophyllene (6.60%), camphane (monoterpenes) (2.95%), Bisabolol (7.31%), tricosane (3.80%), eugenol acetate (3.35%), naphthalene (18.81%), and endo-borneol (19.98%). Certain organic acids such as benzoic acid (33.65%), acetic acid (9.30%), cinnamic acid (6.69%), and phenol (5.47 %). The anti-diabetic activity was evaluated utilizing an α-amylase suppression analysis. The anticancer effect of Cas 3 gene expression analysis by qRT-PCR was assessed in a human bone cancer cell line (MG-63). Clove oil inhibited the α -amylase enzyme concentration-dependently, with an IC50 value (IC50 167.24.46 μg/ml) equivalent to that of conventional acarbose (IC50 85.393.14 μg/ml). The results of real-time qRT-PCR revealed that following treatment with clove oil with an IC50 value of 33.6 μg/ml, the Cas 3 gene expression level in -actin and the human bone cancer cell line (MG-63) was expressed with a fold gene variation difference of 2.07. According to one study, clove oil could be utilized to treat diabetes and human bone cancer.
References
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37. Uthirapathy, S., and Tahir, T. F., 2021 “Withania Somnifera: Correlation of Phytoconstituents with Hypolipidemic and Cardioprotective Activities”, Aro-The Scientific Journal Of Koya University, 9(2), pp. 15-21. doi: 10.14500/aro.10844.
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2. Atawodi, S.E, Atawodi, J.C, Pfundstein, B, Spiegelhalder, B, Bartsch, H, Owen R., 2011. Assessment of the polyphenol components and in vitro antioxidant properties of Syzygium aromaticum (L.) Merr. & Perry. EJEAChe, 10(3), pp 1970– 1978. 3. Atsumi, S., Fujisawa, T., and Tonosaki, K. 2005. Toxicol. In Vitro 19, pp. 1025–1033.
4. Boussaada, O., Chemli, R., 2007. Seasonal variation of essential oil composition of Citrus Aurantium L. J. Essent. Oil-Bear. Plants, 10 (2), pp 109-120.
5. Crawford, E.D., and Wells, J.A., 2011. Caspase substrates and cellular remodeling. Annu Rev Biochem, 80, pp 1055–1087.
6. Enari, M., Sakahira, H., Yokoyama, H., Okawa, K., Iwamatsu, A., Nagata, S., 1998. A caspase-activated Dnase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature, 391, pp 43–50.
7. Ferhat, M.A., Meklati, B.Y., Smadja, J., Chemat, F., 2006. An improved microwave 133ydroalco apparatus for distillation of essential oils from orange peel. J. Chromato. A. 1112 (1-2), pp 121- 126.
8. Hadi, S., 2012. Jurnal Bahan Alam Terbarukan 1, pp 25–30.
9. Hossain, M.A., S. R. Al Harbi, A., M. Weli, Q., Al-Riyami, and Al-Sabahi, J.N., 2014. Asian Pac. J. Trop. Dis, 4, pp 262–268.
10. Huang, X. H, Xiong, P.C, Xiong, C.M, Cai, Y.L, Wei, A.H, Wang, J.P., Liang, X.F, Ruan, J.L., 2010. In vitro and in vivo antitumor activity of Macrothelypteris torresiana and its acute/subacute oral toxicity. Phytomedicine, 17, pp 930–934.
11. Hussain, A., Brahmbhatt, K., Priyani, A., Ahmed, M., Rizvi, T.A., Sharma, C., 2011. Eugenol enhances the chemotherapeutic potential of gemcitabine and induces anticarcinogenic and anti-inflammatory activity in human cervical cancer cells. Cancer Biother. Radiopharm, 26, pp 519–527.
12. Javed Ahamad, Subasini Uthirapathy, Muath Sh. Mohammed Ameen & Esra T. Anwer 2019. Essential Oil Composition and Antidiabetic, Anticancer Activity of Rosmarinus officinalis L. Leaves from Erbil (Iraq). Journal of Essential Oil-Bearing Plants, (22)6, pp.1544-1553, DOI: 10.1080/0972060X.2019.1689179
13. Javed Ahamad., and Subasini Uthirapathy., 2021. Chemical Characterization and Antidiabetic Activity of Essential Oils from Pelargonium graveolens Leaves. ARO journal, http://dx.doi.org/10.14500/aro.10791
14. Jemal, A, Bray, F., Center, M. M., Ferlay, J, Ward, E and Forman D., 2011. Global cancer statistics. CA Cancer J. Clin, 61, pp 69–90.
15. Jirovetz, L., Buchbauer, G., Ngassoum M.B., and Geissler, M., 2002. J. Chromatogr. A 976, pp 265–275.
16. Jirovetz, L., et al., 2006. J. Agric. Food Chem. 54, pp 6303–6307
17. Kaskoos, R.A., Javed Ahamad, and Subasini Uthirapathy., 2021. Chemical Composition and Cytotoxic Activity of Pistacia atlantica var. kurdica Fruits. ARO-The Scientific Journal of Koya University, 9(2), pp. 91-95.
18. Miller GL., 1959. Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem, 31, pp 426–428. Doi: 10.1021/ac60147a030.
19. Neah, L., Ujjwala, M.W., 2011. A unique method for isolation and solubilization of proteins after extraction of RNA from tumor tissue using trizol. J. Biomol. Tech. 22(1), pp 37-44.
20. Park, I.K, Shin, S.C., 2005. Fumigant activity of plant essential oils and components from garlic (Allium sativum) and clove bud (Eugenia caryophyllata) oils against the Japanese termite (Reticulitermes speratus Kolbe) J. Agric. Food Chem, 53, pp 4388–4392.
21. Raj, C.D., Shabi, M.M., Jipnomon, J., R. Dhevi, R., Gayathri, K., Subashini, U., Rajamanickam, G.V., 2012. Terminalia arjuna’s antioxidant effect in isolated perfused kidney. Research in Pharmaceutical Sciences, 7(3), pp. 181-188.
22. Rani, B., Kachhawa, G.R., Yadav, R. K., Chauhan, V. and Raaz, M., 2012. Int. J. Res. Pharmacol. Pharmacother. 1, pp 230–233
23. Ricci, J.E., Munoz-Pinedo, C., Fitzgerald, P., Bailly-Maitre, B., Perkins, G.A., Yadava, N., Scheffler, I.E., Ellisman, M.H., Green, D.R., 2004. Disruption of mitochondrial function during apoptosis is mediated by caspase cleavage of the p75 subunit of complex I of the electron transport chain. Cell, 117, pp 773–786.
24. Saravanan Subramaniam, Ramachandran Subramaniam, Suja Rajapandian, Subasini Uthrapathi, Victor Rajamanickam Gnanamanickam, Govinda Prasad Dubey., 2011. “Anti-Atherogenic Activity of Ethanolic Fraction of Terminalia arjuna Bark on Hypercholesterolemic Rabbits”. Evidence-Based Complementary and Alternative Medicine, vol. 2011, Article ID 487916, 8 pages, 2011. https://doi.org/10.1093/ecam/neq003.
25. Schmitz, J.C, Chu, E., 2011. Effect of small interfering RNA 3’-end overhangs on chemosensitivity to thymidylate synthase inhibitors. Silence, 2(1), pp 88-93.
26. Siagian, V.J., 2014. Outlook Komoditi Cengkeh (Pusat Data dan Sistem Informasi Pertanian Kementrian Pertanian, Jakarta).
27. Sohilait, H.J., 2015. Science Journal of Chemisty 3, pp 95–99. 28. Subasini U, Sundaraganapathy R, Ananda Thangadurai S, Malathi R, Victor Rajamanickam G., 2011. Determination of Nutritive value for certain South Indian Indigenous Species. Int J Pharm & Ind FRes, 1(1), pp. 17-21.
29. Subasini U, Thenmozhi S, Sathyamurthy D, Vetriselvan S, Rajamanickam GV, et al. 2013. Pharmacognostic and phytochemical investigations of Dioscorea bulbifera L. Int J Pharm Life Sci 4, pp. 2693-2700
30. Subasini Uthirapathy, and Javed Ahamad, 2019. Phytochemical Analysis of different fractions of Terminalia Arjuna bark by GCMS, Int. Res. J. Pharm, 10(1), pp 42-48.
31. Tahsin, T., & Arif, A., & Bayrakdar, B, & Uthirapathy, S., 2022. In-Vitro Antioxidant Activity of Avocado Fruit Oil in Erbil, Kurdistan Region. Eurasian Journal of Science and Engineering, 8(3),298-307
32. Tamilarasi, C.T, Subasini, U, Kavimani, S, Jaykar, B., 2000. Phytochemical and Pharmacological Evaluation of Ampelocissus latifolia. Ancient Sci Life, 20(1), pp. 1-6
33. Thenmozhi S, Subasini U, Kameshwaran S, Dhanalakshmi M, Rajamanickam G.V., 2013. Morpho-anatomical and preliminary phytochemical studies of leaves of Gynandropsis pentaphylla Linn. Int J of Pharm & Life Sci, 4(7), pp. 2800-2809.
34. Thenmozhi S. and Subasini U., 2016. Isolation, characterization and in-vitro cytotoxic study of vitexin from Vitex pinnata Linn. leaves. IJRPP, 1, pp. 84–89.
35. Uthirapathy, S. (2023) “Cytostatic Effects of Avocado Oil Using Single-cell Gel Electrophoresis (Comet Assay): An
Evaluation”, Aro-The Scientific Journal Of Koya University, 11(1), pp. 16-21. doi: 10.14500/aro.11073.
36. Uthirapathy, S., 2019. Novel biomarkers of atherogenic diet induced dyslipidemia and metabolic syndrome suppressed by Terminalia arjuna. Int J Pharma Sci Res, 10, pp.2528–2536. https://doi.org/10.13040/IJPSR.0975-8232.10(5).2528-36.
37. Uthirapathy, S., and Tahir, T. F., 2021 “Withania Somnifera: Correlation of Phytoconstituents with Hypolipidemic and Cardioprotective Activities”, Aro-The Scientific Journal Of Koya University, 9(2), pp. 15-21. doi: 10.14500/aro.10844.
38. Uthirapathy, Subasini and Ahamad, Javed., 2022. Toxicity profiles of the Hydroalcoholic seed extract of Psoralea Corylifolia L Fabaceae in Wistar rats. Academic Journal of Health Sciencies Medicina balear, 37 (3), pp. 120-126
39. Uthirapathy, Subasini and Tahsin, Amani., 2021. Evaluation of Genotoxic effects of a Hydro-alcoholic extract of flowers of Nargis (Narcissus Tazetta L.). Eurasian Journal of Science & Engineering, 7 (2). Pp. 39-48.
40. WHO Fact Sheet Fact sheet. 2015. N°312 http://www.who.int/mediacentre/factsheets/fs312. Accessed 24 Jan 2016
41. Wyld, L, Reed, M., 2007. The role of surgery in the management of older women with breast cancer. Eur. J. Cancer, 43, pp 2253–2263.
42. Siagian, V.J., 2014. Outlook Komoditi Cengkeh (Pusat Data dan Sistem Informasi Pertanian Kementrian Pertanian, Jakarta).
43. Sohilait, H.J., 2015. Science Journal of Chemisty 3, pp 95–99.
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May 13, 2023
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Ali Omar Yassen, Rozhan Arif, & Subasini Uthirapathy. (2023). SYBR-Green I fluorescence-based qRT-PCR of clove oil’s anticancer, chemical characterization and antidiabetic activities. Journal of Population Therapeutics and Clinical Pharmacology, 30(11), 125–134. https://doi.org/10.47750/jptcp.2023.30.11.015
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