Yasir IHTESHAM, Uzma KHAN, ZEYNEP DOĞAN, VAHAP MURAT KUTLUAY, İCLAL SARAÇOĞLU

Evaluation of Some Biological Effects of Incarvillea emodi (Royle ex Lindl.) Chatterjee and Determination of Its Active Constituents

This present study was an evaluation of the antioxidant and cytotoxic activities of Incarvillea emodi (Bignoniaceae). The aqueous extracts of different parts of Incarvillea emodi, collected from different places in Pakistan, were tested for radical scavenging effects on 2,2-diphenyl-1-picrylhydrazil (DPPH), nitric oxide (NO), superoxide (SO) and [2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)] (ABTS•+) radicals. DPPH radical scavenging effects of three polyamide column fractions of one extract were also tested. The cytotoxicity of the extracts were tested against Hep-2 (human larynx epidermoid carcinoma) cancer cell line by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide] method. In addition, an HPLC-DAD system was used to show the presence of phenylethanoid glycosides in the most active polyamide column fraction. The free radical scavenging effects of the extracts were found comparable to that of reference antioxidants, 3-t-butyl-4-hydroxyanizole (BHA), quercetin and ascorbic acid (AA). Concentration dependent cytotoxic activity was observed against Hep-2 cancer cell line. Two phenylethanoid glycosides, acteoside and leucosceptoside A, were identified in the active polyamide column fraction at HPLC-DAD system. The presence of phenylethanoid glycosides in Incarvillea emodi was shown for the first time with this study. Antioxidant and cytotoxic activity researches are important for developing new drugs. Our results supported to use of Incarvillea emodi as folk medicine due to several biological effects in Pakistan. Moreover, the use of Incarvillea genus as a traditional ethnoveterinary medicine in dyspepsia and internal diseases were reported in previous studies.

Incarvillea emodi (Royle ex Lindl.) Chatterjee’nin Bazı Biyolojik Etkilerinin Değerlendirilmesi ve Aktif Bileşiklerinin Tayini

Bu çalışmada, Incarvillea emodi’nin antioksidan ve sitotoksik etkileri araştırılmıştır. Pakistan’ın farklı bölgelerinden toplanan Incarvillea emodi’nin farklı kısımlarından hazırlanan sulu ekstrelerin radikal süpürücü etkileri, 2,2-difenil-1-pikrilhidrazil (DPPH), nitrik oksit (NO), süperoksit (SO) ve 2,2’-azino-bis 3-etilbenzotiyazolin-6-sülfonik asit (ABTS•+) radikallerine karşı incelenmiştir. Bir ekstrenin üç poliamit kolon fraksiyonunun DPPH radikal süpürücü etkileri de araştırılmıştır. Ekstrelerin sitotoksik aktiviteleri, MTT [3-(4,5-dimetiltiyazol-2-il)- 2,5-difeniltetrazolyum bromür] yöntemi kullanılarak Hep-2 (insan larinks epidermoit karsinoma) kanser hücre dizisine karşı saptanmıştır. Ayrıca, en etkili poliamit kolon fraksiyonunda, feniletanoit glikozitlerinin varlığını göstermek için bir HPLC-DAD sistemi geliştirilmiştir. Ekstrelerin serbest radikal süpürücü etkileri, referans antioksidanlar, 3-t-butil-4-hidroksianizol (BHA), kersetin ve askorbik asit (AA) ile karşılaştırılabilir seviyede bulunmuştur. Hep-2 kanser hücre dizisine karşı konsantrasyona bağlı sitotoksik aktivite gözlenmiştir. HPLCDAD sistemine uygulanmış etkili poliamit kolon fraksiyonunda, akteozit ve lökoseptozit A isimli 2 feniletanoit glikozitinin bulunduğu tespit edilmiştir. Incarvillea emodi’de feniletanoit glikozitlerinin varlığı ilk kez bu çalışma ile gösterilmiştir. Antioksidan ve sitotoksik aktivite araştırmaları ilaç geliştirme çalışmalarında önemlidir. Çalışmamızın sonuçları, Incarvillea emodi’nin, Pakistan’da çeşitli biyolojik etkileri nedeniyle halk arasında kullanılışını desteklemektedir. Ayrıca, Incarvillea cinsinin dispepsi ve iç hastalıklarda geleneksel etnoveteriner ilaç olarak kullanımı da daha önceki çalışmalarda belirtilmektedir.

PDF

___

Bernstein N, Akram M, Daniyal M, Koltai H, Fridlender M, Gorelick J: Chapter Four- Antiinflammatory potential of medicinal plants: A source for therapeutic secondary metabolites. Adv Agron, 150, 131-183, 2018. DOI: 10.1016/bs.agron.2018.02.003

Gupta SS: Prospects and perspectives of natural plant products in medicine. Indian J Pharmacol, 26 (1): 1-12, 1994.

Edeoga HO, Okwu DE, Mbaebie BO: Phytochemical constituents of some Nigerian medicinal plants. Afr J Biotechnol, 4 (7): 685-688, 2005. DOI: 10.5897/AJB2005.000-3127

Barnes S: Effect of genistein on in vitro and in vivo models of cancer. J Nutr, 125 (Suppl. 3): 777S-783S, 1995.

Bilir EK, Tutun H, Sevin S, Kısmalı G, Yarsan E: Cytotoxic effects of Rhododendron ponticum L. extract on prostate carcinoma and adenocarcinoma cell line (DU145, PC3). Kafkas Unıv Vet Fak Derg, 24 (3): 451-457, 2018. DOI: 10.9775/kvfd.2017.19219

Demirkan I, Ertekin T, Korkmaz M, Kılıc I, Cevik-Demirkan A, Bozkurt F, Konuktürk A: Antitumorigenic activity of the herbal mixtureAK27 on ehrlich ascites carcinoma in mice. Kafkas Unıv Vet Fak Derg, 23 (5): 817-823, 2017. DOI: 10.9775/kvfd.2017.17971

Chen S, Guan K, Zhou Z, Olmstead R, Cronk Q: Molecular phylogeny of Incarvillea (Bignoniaceae) based on ITS and TRN L-F sequences. Am J Bot, 92 (4):625-633, 2005. DOI: 10.3732/ajb.92.4.625

Ahmad H, Ahmad A, Jan MM: The medicinal plants of salt range, J Biol Sci, 2 (3): 175-177, 2002. DOI: 10.3923/jbs.2002.175.177

Shang X, Tao C, Miao X, Wang D, Dawa T, Wang Y, Yang Y, Pan H: Ethno-veterinary survey of medicinal plants in Ruoergai region, Sichuan province, China, J Ethnopharmacol, 142, 390-400, 2012. DOI: 10.1016/j. jep.2012.05.006

Fu JJ, Jin HZ, Shen YH, Qin JJ, Wang Y, Huang Y, Zeng Q, Zhang WD: Chemical constituents of plants from the genus Incarvillea. Chem Biodivers, 6 (6): 818-826, 2009. DOI: 10.1002/cbdv.200800166

Chen S, Gong J, Guan K, Zhou Z: Biodiversity conservation of the genus Incarvillea Juss. (Bignoniaceae) based on molecular diversity and species richness assessment. J Plant Biol, 53, 387-394, 2010. DOI: 10.1007/ s12374-010-9127-6

Rana A, Dhyani D, Gulati A, Singh HP: Isolation of two major iridoid glucosides from Incarvillea emodi. Nat Prod Res, 25 (10): 1014-1017, 2011. DOI: 10.1080/14786419.2010.523003

Yu ZW, Zhu HY, Yang XS, Sun QY, Hao XJ: Study on chemical constituents from Incarvillea arguta and their accelerating PC-12 cell differentiation. Zhongguo Zhong Yao Za Zhi, 30 (17): 1335-1338, 2005.

Yuan CS, Zhang Q, Xie WD, Yang XP, Jia ZJ: Iridoids from Pedicularis kansuensis forma albiflora. Pharmazie, 58, 428-430, 2003. DOI: 10.1002/ chin.200340174

Harput US, Genc Y, Saracoglu I: Cytotoxic and antioxidative activities of Plantago lagopus L. and characterization of its bioactive compounds. Food Chem Toxicol, 50 (5): 1554-1559, 2012. DOI: 10.1016/j. fct.2012.01.019

Jensen SR, Gotfredsen CH, Harput US, Saracoglu I: Chlorinated iridoid glucosides from Veronica longifolia and their antioxidant activity. J Nat Prod, 73 (9):1593-1596, 2010. DOI: 10.1021/np100366k

Hensley K, Mou S, Pye QN: Nitrite determination by colorimetric and fluorometric griess diazotization assays. In, Hensley K, Floyd RA (Eds): Methods in Biological Oxidative Stress. 185-193, Humana Press Inc, Totowa, 2003.

Harput US, Genc Y, Khan N, Saracoglu I: Radical scavenging effects of different Veronica species. Rec Nat Prod, 5 (2): 100-107, 2011.

Elizabeth K, Rao MNA: Oxygen radical scavenging activity of curcumin. Int J Pharm, 58, 237-240, 1990. DOI: 10.1016/0378-5173(90)90201-E

Re R, Pellegrini N, Proteggente A, Pannala A, Yang M, RiceEvans C: Antioxidant activity applying an improved ABTS radical cation decolorization assay. Free Radic Biol Med, 26 (9-10): 1231-1237, 1999. DOI: 10.1016/S0891-5849(98)00315-3

Saracoglu I, Inoue M, Calis I, Ogihara Y: Studies on constituents with cytotoxic and cytostatic activity of two Turkish medicinal plants Phlomis armeniaca and Scutellaria salviifolia. Biol Pharm Bull, 18, 1396-1400, 1995. DOI: 10.1248/bpb.18.1396

Rana A, Rana S, Majeed R, Singh HP, Gulati A, Hamid A, Vyas D, Dhyani D: Comparative studies for screening of bioactive constituents from various parts of Incarvillea emodi. Nat Prod Res, 28 (8): 593-596, 2014. DOI: 10.1080/14786419.2014.886207

Mascarenhas M: Structure-activity characterization, a quick method to screen mushrooms for the presence of antitumor glucans. Mushroom Res, 3 (2): 77-80, 1994.

Ihtesham Y, Dogan Z, Kutluay VM, Khan U, Saracoglu I: Bioassay guided isolation studies on Incarvillea emodi (Royle ex Lindl) Chatterjee. Planta Med, 82 (S 01): S1-S381, 2016. DOI: 10.1055/s-0036-1596576