PHYTOCHEMICAL STUDY AND THE ANTIPROLIFERATIVE ACTIVITY OF INULA VULGARIS SPECIES GROWN IN LEBANON
Objective: Cancer represents the second leading cause of death after stroke and heart diseases. Plant extracts have long been used in traditional medicine for the prevention and treatment of many illnesses, including some types of cancer. The aim of this study was to evaluate the antiproliferative effects of ethyl acetate fractions of two Lebanese herbs: Inula viscosa (I. vis) and Inula vulgaris (I. vul).
Methods: Plants were extracted with ethanol followed by ethyl acetate, then dried and tested on three cell lines including CaCO2, HepG2, and MCF7, to check for their viability and antiproliferative activity, using trypan blue exclusion and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays. Proton (1H) and carbon (13C) nuclear magnetic resonance spectrometry (NMR) were used to identify the compounds that have been isolated from both Inula species.
Results: The current findings were consistent for both trypan blue and MTT assays. The results showed that the most potent effect for I. vul was HepG2 (IC50 20 Âµg/ml, 27 Âµg/ml), and for I. vis on MCF7 (9 Âµg/ml, 15 Âµg/ml) and CaCO2 (12 Âµg/ml, 22 Âµg/ml) in the two mentioned assays respectively. However, insignificant differences were observed among the studied plants for each of the evaluated cells indicating comparable potencies. Quercetin, quercetin glycoside, and epicatechin derivatives were isolated by fractionation on column chromatography and identified using NMR spectroscopy.
Conclusion: The antiproliferative activities of the two plants could be related to their content that is significant for high levels of secondary metabolites. The identification of those compounds is necessary to establish a relationship between their chemical structures and their activities.
2. Tohme G, Tohme H. Illustrated Flora of Lebanon. Lebanese National Council for Scientific Research; 2007.
3. Manez S, Hernandez V, Giner RM, Rios JL, Recio M. Inhibition of pro-inflammatory enzymes by Inu-Visco Lide, a sesquiterpene lactone from I. vis. Fitoterapia 2007;78:329-31.
4. Dom DC, Alexenizer MH, Engstler JG, Dom A. Tumor cell specific toxicity of I. helenium extracts. Phytother Res 2006;20:970-80.
5. Assi M, Aboul-Ela M, El-Lakany A. A comparative phytochemical and antimicrobial study of some Inula species Growing in Lebanonâ€, MS thesis, BAU; 2011.
6. Nesterova IuV, Zelenskaia KL, Vetoshkina TV, Aksinenko SG, Gorbacheva AV, Gorbatykh NA. Mechanisms of antistressor activity of Inula helenium preparations. Eksp Klin Farmakol 2003;66:63-5.
7. Zhang H, Wen J, Wang Y, Zheng B, Han M. Flavonoids from Inula britannica inhibit injury-induced neointimal formation by suppressing oxidative-stress generation. J Ethnopharmacol 2009;126:176-83.
8. Aboul-Ela M, El lakany A, Abdel-Kader M, Alqasoumi S, Shamseddine S, Hammoda HM. New quinic acid derivatives from hepatoprotective I. crithmoides root extract. Helv Chim Acta 2012;95:61-6.
9. Zelenskaja KL, Povetjeva TN, Pashinskij VG. Anesthetic effect of alcohol extractions from Inula helenium. Rastit Resur 2003;39:82-6.
10. Torre LA, Rebecca LFB, Jacques S, Joannie F, Tieulent L, Jemal A. Global cancer statistics. CA Cancer J Clin 2015;65:87-108.
11. Shamseddine A. Cancer trends in Lebanon: a review of incidence rates for the period of 2003â€“2008 and projections until 2018. Population Health Metrics 2014;12:4.
12. Hartwell JL. Plants used against cancer: a survey. Lawrence MA. Quarterman Publications; 1982;438-9.
13. Cordell G. Natural products in drug discoveryâ€“creating a new vision. Phytochem Rev 2002;1:261â€“73.
14. Paul M, BrÃ¼ning G, Bergmann J, Jauch J. A thin-layer chromatography method for the identification of three different olibanum resins (Boswellia serrata, Boswellia papyrifera and Boswellia carterii, respectively, Boswellia sacra) Phytochem Anal 2012;23:184-9.
15. Rodrigueg J, Yanez J, Vicente V, Alcaraz M, Benavente-GarcÃa O, Lazona JA, et al. Effects of several flavonoids on the growth of B16f10 and SK-MEL-1 melanoma cell lines: the relationship between structure and activity. Melanoma Res 2002;12:99-107.
16. Uzair M. Phytochemical and biological studies of C. bonarenesis, Euphorbia prostata and Euphorebia helioscopiaâ€ PhD thesis, Bahauddin Zakariya University, Pakistan; 2009.
17. Mahmouda A, Al-Shihrya SS, Hegazy MF. The biological activity of a phloroglucinol glucoside derivative from Conyza aegyptiaca. Natl Res Centre 2009;64:513â€“7.
18. Sanjay P, Nirav G, Ashok S, Anand S. In vitro cytotoxicity activity of Solanum Nigrum extract against HELA cell line and VERO cell line. Int J Pharm Pharm Sci 2009;1:38-45.
19. Alley MC, Scudiero DA, Monks A, Hursey ML, Czerwinski MJ, Fine DL, et al. Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res 1988;48:589â€“601.
20. Lau C, Ho C, Kim C, Leung K, Fung K, Tse T, et al. Cytotoxic activities of Coriolus versicolor (Yunzhi) extract on human leukemia and lymphoma cells by induction of apoptosis. Life Sci 2004;75:797â€“808.
21. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 1983;65:55-63.
22. Saikali M, Ghantous A, Halawi R, Talhouk SN, Saliba N, Darwiche N. Sesquiterpene lactones isolated from indigenous middle eastern plants inhibit tumor promotor-induced transformation of JB6 cells. BMC Complementary Altern Med 2012;12:89.
23. Cai Y, Davies AP, Lewis JR. 1H and 13C NMR assignments of some green tea polyphenols. Magn Reson Chem 1996;34:887-90.
24. Itharat A, Houghton PJ, Eno-Amooquaye E, Burke PJ, Sampson JH, Raman A. In vitro cytotoxic activity of Thai medicinal plants used traditionally to treat cancer. J Ethnopharmacol 2004;90:33â€“8.
25. MoffoK S, Haba H, Lavaud C, Long C, Benkhaled M. Chemical constituents of Centaurea omphalotricha. Rec Nat Prod 2012;6:292-5.
26. El-Lakany A, Aboul-Ela M, Hammouda HM, Ghazi NM, Mahmoud Z. New Methoxylated flavonols from I. crithmoides L. Die Pharmazie 1996;51:435-6.
27. El-Lakany A, Aboul-Ela M, Hammouda HM, Abdul-Ghani M. Three new butyl glycosides from I. crithmoides L. growing in Egypt.
Die Phrmazie 2003;58:940-2.
28. Ren W. Flavonoids: promising anticancer agents. Med Res Rev 2003;23:519-34.
29. Messaoudi M. Cytotoxic effect and chemical composition of I. vis from three different regions of morocco. Eur J Med Plants 2016;16:1-9.
30. Claudia KR. Epicatechin gallate and catechin gallate are superior to epigallocatechin gallate in growth suppression and anti-inflammatory activities in pancreatic tumor cells. Cancer Sci 2011;102:728-34.