Cytotoxic and Antioxidant Activities of Crocus pallasii subsp. haussknechtii Corms Extracts Compared with Crocus sativus

Document Type: Original paper

Authors

1 Department of Biological Science and Biotechnology, Faculty of Science, University of Kurdistan, Sanandaj, Iran.

2 Department of Horticultural Science and Engineering, Research Center of Medicinal Plants Breeding and Development, University of Kurdistan, Sanandaj, Iran.

3 Immunoregulation Research Center, Shahed University, Tehran, Iran.

4 Institute of Biochemistry and Biophysics, University of Tehran, Tehran, Iran.

5 Department of Biotechnology, Iranian Research Organization for Science and Technology, Tehran, Iran.

Abstract

Background and objectives: The goal of this study was to compare the cytotoxic and antioxidant activities of the corms extracts of two Crocus species (C. sativus L. and C. pallasii subsp. haussknechtii). Methods: n-Hexan, chloroform and methanol extracts of the corms were prepared. Cytotoxic activities of the extracts against human breast cancer cell lines MDA-MB-231 and MCF-7 were evaluated via MTT assay and dual acridine orange/ethidium bromide fluorescent staining test. Antioxidant activities of the extracts were assessed by DPPH method. Results: The results showed that the methanol extracts of the corms from both species had cytotoxic activity against the breast cancer cells. Cytotoxic activity of the corms extract of C. sativus L. was significantly higher than C. pallasii subsp. haussknechtii. n-Hexan and chloroform extracts from the corms of C. pallasii subsp. haussknechtii and Crocus sativus L. showed no anti-proliferative activity which indicated the polar nature of the cytotoxic agent(s) in the extracts. The results of dual acridine orange/ethidium bromide fluorescent staining test revealed that the methanol extracts of both species corms destroyed cancer cells through apoptosis. Moreover, the methanol corm extracts of the two species showed little antioxidant activity. Conclusion: It can be concluded that C. sativus L. may possess more significant apoptotic properties compared to C. pallasii subsp. haussknechtii.

Keywords


[1] DeSantis CE, Ma J, Sauer AG, Newman LA, Jemal A. Breast cancer statistics, 2017, racial disparity in mortality by state. CA Cancer J Clin. 2017; 67(6): 439-448.

[2] Goldman B. Multidrug resistance: can new drugs help chemotherapy score against cancer? J Natl Cancer Inst. 2003; 95(4): 255-257.

[3] Balunas MJ, Kinghorn AD. Drug discovery from medicinal plants. Life Sci. 2005. 78(5): 431-441.

[4] Khorasanchi Z, Shafiee M, Kermanshahi F, Khazaei M, Ryzhikov M, Parizadeh MR, Kermanshahi B, Ferns GA, Avan A, Hassanian SM. Crocus sativus a natural food coloring and flavoring has potent anti-tumor properties. Phytomed. 2018; 43: 21-27.

[5] Sanei M, Rahimyan H, Agayev Y, Soheilivand S. New cytotype of Crocus pallasii subsp. haussknechtii from west of Iran. Acta Hortic. 2007; 739: 107-111.

[6] Mosaddegh M, Esmaeili S, Eslami-tehrani B, Kermatian B, Mohebby S, Hamzeloo-moghadam M. Anti-angiogenesis properties of Crocus pallasii subsp. haussknechtii, a popular ethnic food. Res J Pharmacogn. 2015; 2(3): 37-42.

[7] Hamzeloo-moghadam M, Shahrestani R, Keramatian B, Mohebby S. Cytotoxic activity of Crocus pallasii subsp. haussknechtii in human cancer cell lines. Int J Pharm Sci Res. 2019; 10(1): 117-120.

[8] Azmir J, Zaidul I, Rahman M, Sharif K, Mohamed A, Sahena F, Jahurul M, Ghafoor K, Norulaini N, Omar A. Techniques for extraction of bioactive compounds from plant materials: a review. J Food Eng. 2013; 117(4): 426-436.

[9] Van Meerloo J, Kaspers GJ, Cloos J. Cell sensitivity assays: the MTT assay. Methods Mol Biol. 2011; 731: 237-245.

[10] Smith SM, Ribble D, Goldstein NB, Norris DA, Shellman YG. A simple technique for quantifying apoptosis in 96-well plates. Methods Cell Biol. 2012; 112: 361-368.

[11] Sharma OMP, Bhat TK. DPPH antioxidant assay revisited. Food Chem. 2009; 113(4): 1202-1205.

[12] Escribano J, Dı́az-guerra MJM, Riese HH, Ontañón J, Garcı́a-olmo D, Garcı́a-olmo DC, Rubio A, Fernández JA. In vitro activation of macrophages by a novel proteoglycan isolated from corms of Crocus sativus L. Cancer Lett. 1999; 144(1): 107-114.

[13] Escribano J, Rı́os I, Fernández JA. Isolation and cytotoxic properties of a novel glycoconjugate from corms of saffron plant (Crocus sativus L.). Biochim Biophys Acta Gen Subj. 1999; 1426(1): 217-222.

[14] Escribano J, Diaz-guerra M, Riese HH, Alvarez A, Proenza R, Fernández JA. The cytolytic effect of a glycoconjugate extracted from corms of saffron plant (Crocus sativus) on human cell lines in culture. Planta Med. 2000; 66(2): 157-162.

[15] Shakeri R, Kheirollahi A, Davoodi J. Apaf-1: Regulation and function in cell death. Biochimie. 2017; 135: 111-125.

[16] Hengartner MO. The biochemistry of apoptosis. Nature. 2000; 407(6805): 770-776.

[17] Johnstone RW, Ruefli AA, Lowe SW. Apoptosis: a link between cancer genetics and chemotherapy. Cell. 2002; 108(2): 153-164.

[18] Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983; 65(1-2): 55-63.

[19] Liu K, Liu PC, Liu R, Wu X. Dual AO/EB staining to detect apoptosis in osteosarcoma cells compared with flow cytometry. Med Sci Monit Basic Res. 2015; 21: 15-20.

[20] Vermes I, Haanen C, Reutelingsperger C. Flow cytometry of apoptotic cell death. J Immunol Methods. 2000; 243(1-2): 167-190.

[21] Zhang Y, Chen X, Gueydan C, Han J. Plasma membrane changes during programmed cell deaths. Cell Res. 2018; 28(1): 9-21.

[22] Kasibhatla S, Amarante-mendes GP, Finucane D, Brunner T, Bossy-wetzel E, Green DR. Acridine orange/ethidium bromide (AO/EB) staining to detect apoptosis. CSH Protoc. 2006; Article ID pdb.prot4493. 

[23] Esmaeili N, Ebrahimzadeh H, Abdi K, Safarian S. Determination of some phenolic compounds in Crocus sativus L. corms and its antioxidant activities study. Pharmacogn Mag. 2011; 7(25): 74-80.

[24] Baba SA, Malik AH, Wani ZA, Mohiuddin T, Shah Z, Abbas N, Ashraf N. Phytochemical analysis and antioxidant activity of different tissue types of Crocus sativus and oxidative stress alleviating potential of saffron extract in plants, bacteria, and yeast. S Afr J Bot. 2015; 99: 80-87.

[25] Barchan A, Bakkali M, Arakrak A, Pagán R, Laglaoui A. The effects of solvents polarity on the phenolic contents and antioxidant activity of three Mentha species extracts. Int J Curr Microbiol App Sci. 2014; 3(11): 399-412.

[26] Dorman HD, Koşar M, Kahlos K, Holm Y, Hiltunen R. Antioxidant properties and composition of aqueous extracts from Mentha species, hybrids, varieties, and cultivars. J Agric Food Chem. 2003; 51(16): 4563-4569.