TY  - JOUR
T1  - In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts
AU  - Karadağ, Ayşe Esra
AU  - Çaşkurlu Öz, Ayşegül
AU  - Karavuş, Şule Nur
AU  - Biltekin, Sevde Nur
AU  - Helvaci, Esra Zeynep
PY  - 2025
DA  - July
JF  - Journal of Research in Pharmacy
JO  - J. Res. Pharm.
PB  - Marmara University
WT  - DergiPark
SN  - 2630-6344
SP  - 2115
EP  - 2123
VL  - 27
IS  - 5
LA  - en
AB  - The aim of this study was to evaluate phytochemical composition of Ajuga reptans L. (Lamiaceae) aerial parts (separately flower and leaf) methanol, aqueous-methanolic extracts, and their cytotoxic activities. The phytochemical analysis was performed high-performance liquid chromatography (HPLC). Caffeic acid, p-coumaric, gallic, chlorogenic, ferulic acids, kaempferol, rutin, quercetin, quercetin-3-O-galactoside, and quercitrin were used as reference substances by HPLC in all samples. The major compounds in the extract were found as ferulic acid, caffeic acid, rutin, and quercetin-3-O-galactoside. Cytotoxicity was investigated using methyl thiazole tetrazolium (MTT) assay. Cytotoxic evaluation of the extracts against cancer (MCF7, PC3, and A549) and healthy human embryonic kidney cell line (HEK293) cell lines by MTT. Compared to other cells, the methanol extract of A. reptans demonstrated high selectivity against PC3 cells (IC50: 95  0.99 μg/mL) and selectivity index was four times higher than reference drug colchicine. (IC50: 95  0.99 μg/mL, SI: 6.10). A. reptans demonstrated antiproliferative potential against prostate and lung cancer cells. Therefore, additional investigations are needed to study the mechanism of the cytotoxicity for A. reptans.
KW  - Ajuga
KW  - Cytotoxicity
KW  - Lamiaceae
KW  - HPLC
CR  - [1] Sadati N, Jenett-Siems K, Siems K, Ardekania MRS, Hadjiakondii A, Akbarzadeh T, Ostad ZN, Khanavi M. Major constituents and cytotoxic effects of Ajuga chamaecistus ssp. tomentella. Z Naturforsch C. 2012; 67(5-6): 275-281. https://doi.org/10.1515/znc-2012-5-606
CR  - [2] Ghita G, Cioanca O, Gille E, Necula R, Zamfirache MM, Stanescu U. Contributions to the phytochemical study of some samples of Ajuga reptans L. and Ajuga genevensis. Bull Transilvania University of Braov Series VI: Medical Sci. 2011; 4(53):2-2011.
CR  - [3] Zafar HI, Badiaa L. Ethnopharmacology of the plants of genus Ajuga. Pakistan J Pharm Sci. 2009; 22(4): 425-462.
CR  - [4] Toiu A, Vlase L, Gheldiu AM, Vodnar D, Oniga I. Evaluation of the antioxidant and antibacterial potential of bioactive compounds from Ajuga reptans extracts. Farmacia. 2017; 65(3): 351-355.
CR  - [5] Khare CP. Indian Medicinal Plants-An Illustrated Dictionary. 1st Indian Reprint, Springer (India). 2007; 28.
CR  - [6] Calcagno MP, Camps F, Coil J, Mele E, Sánchez-Baeza F. New phytoecdysteroids from roots of Ajuga reptans varieties. Tetrahedron. 1996; 52(30): 10137-10146. https://doi.org/10.1016/0040-4020(96)00536-4
CR  - [7] Shoji N, Umeyama A, Sunahara N, Arihara S. Ajureptoside, a novel C9 iridoid glucoside from Ajuga reptans. J Nat Prod. 1992; 55(7): 1004-1006. https://doi.org/10.1021/np50085a030
CR  - [8] Carbonell P, Coll J. Ajugatansins, neo-clerodane diterpenes from Ajuga reptans. Phytochem Anal. 2001; 12(1): 73-78. https://doi.org/10.1002/1099-1565(200101/02)12:1 <73::AID-PCA561>3.0.CO;2-8
CR  - [9] Malakov PY, Papanov GY, Perales A, Maria C, Rodríguez B. The absolute stereochemistry of ajugavensins, neo-clerodane diterpenes from Ajuga genevensis: a revision of the C-1 configuration of Ajugavensins A and B. Phytochem. 1992; 31(9): 3151-3153. https://doi.org/10.1016/0031-9422(92)83464-A
CR  - [10] Terahara N, Callebaut A, Ohba R, Nagata T, Ohnishi-Kameyama M, Suzuki M. Triacylated anthocyanins from Ajuga reptans flowers and cell cultures. Phytochem. 1996; 42(1): 199-203. https://doi.org/10.1016/0031-9422(95)00838-1
CR  - [11] Pal A, Toppo FA, Chaurasiya PK, Singour PK, Pawar RS. In-vitro cytotoxicity study of methanolic fraction from Ajuga bracteosa Wall ex. Benth on MCF-7 breast adenocarcinoma and hep-2 larynx carcinoma cell lines. Pharmacognosy Res. 2014; 6(1): 87-91. https://doi.org/10.4103/0974-8490.122923
CR  - [12] Rubnawaz S, Okla MK, Akhtar N, Khan IU, Bhatti MZ, Dhong HQ, El-Tayeb MA, El-badawi YB, Almaary KS, Moussa IM, Abbas ZK, Mirza B. Antibacterial, antihemolytic, cytotoxic, anticancer, and antileishmanial effects of Ajuga bracteosa transgenic plants. Plants. 2021; 10(9): 1894. https://doi.org/10.3390/plants10091894
CR  - [13] Zahra SS, Ahmed M, Qasim M, Zia M, Mirza B, Haq I. Polarity based characterization of biologically active extracts of Ajuga bracteosa Wall. ex Benth. and RP-HPLC analysis. BMC Complement Altern Med. 2017; 17(1): 1-16. https://doi.org/10.1186/s12906-017-1951-5
CR  - [14] Okur ME, Karakaş N, Karadağ AE, Yılmaz R, Demirci F. In vitro cytotoxicity evaluation of Marrubium vulgare L. methanol extract. J Res Pharm. 2019; 23(4): 711-718. http://doi.org/10.12991/jrp.2019.180
CR  - [15] Oran SA, Althaher AR, al Shhab MA. Chemical composition, in vitro assessment of antioxidant properties and cytotoxicity activity of ethanolic and aqueous extracts of Ajuga orientalis L. (Lamiaceae). J Pharm Pharmacogn Res. 2022; 10(3): 486-495. https://doi.org/10.56499/jppres22.1344_10.3.486
CR  - [16] Rauca VF, Vlase L, Casian T, Sesarman A, Gheldiu AM, Mocan A, Banciu M, Toiu A. Biologically active Ajuga species extracts modulate supportive processes for cancer cell development. Front Pharmacol. 2019; 10: 334. https://doi.org/10.3389/fphar.2019.00334
CR  - [17] 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. https://doi.org/10.1016/0022-1759(83)90303-4
CR  - [18] Vlaisavljević S, Šibul F, Sinka I, Zupko I, Ocsovszki I, Jovanović-Šanta S. Chemical composition, antioxidant and anticancer activity of licorice from Fruska Gora locality. Ind Crops Prod. 2018; 112: 217-224. https://doi.org/10.1016/j.indcrop.2017.11.050
CR  - [19] Imani A, Maleki N, Bohlouli S, Kouhsoltani M, Sharifi S, Maleki Dizaj S. Molecular mechanisms of anticancer effect of rutin. Phytother Res. 2021; 35(5): 2500-2513. https://doi.org/10.1002/ptr.6977
CR  - [20] Satari A, Ghasemi S, Habtemariam S, Asgharian S, Lorigooini Z. Rutin: a flavonoid as an effective sensitizer for anticancer therapy; Insights into multifaceted mechanisms and applicability for combination therapy. Evid Based Complement Altern Med. 2021; 2021: 9913179. https://doi.org/10.1155/2021/9913179
CR  - [21] Alonso-Castro AJ, Domínguez F, García-Carrancá A. Rutin exerts antitumor effects on nude mice bearing SW480 tumor. Arch Med Res. 2013; 44(5): 346-351. https://doi.org/10.1016/j.arcmed.2013.06.002
CR  - [22] Agmon E, Solon J, Bassereau P, Stockwell BR. Modeling the effects of lipid peroxidation during ferroptosis on membrane properties. Sci Rep. 2018; 8(1): 1-11. https://doi.org/10.1038/s41598-018-23408-0
CR  - [23] Gutteridge JM. Lipid peroxidation and antioxidants as biomarkers of tissue damage. Clin Chem. 1995; 41(12): 1819-1828. https://doi.org/10.1093/clinchem/41.12.1819
CR  - [24] Patel K, Patel DK. The beneficial role of rutin, a naturally occurring flavonoid in health promotion and disease prevention: a systematic review and update. Bioactive Food as Dietary Interventions for Arthritis and Related Inflammatory Diseases. 2019: 457-479. https://doi.org/10.1016/B978-0-12-813820-5.00026-X
CR  - [25] Abotaleb M, Liskova A, Kubatka P, Büsselberg D. Therapeutic potential of plant phenolic acids in the treatment of cancer. Biomolecules. 2020; 10(2): 221-226. https://doi.org/10.3390/biom10020221
CR  - [26] Huang MT, Ma W, Yen P, Xie JG, Han J, Frenkel K, Conney AH. Inhibitory effects of caffeic acid phenethyl ester (CAPE) on 12-O-tetradecanoylphorbol-13-acetate-induced tumor promotion in mouse skin and the synthesis of DNA, RNA and protein in HeLa cells. Carcinogenesis. 1996; 17(4): 761-765. https://doi.org/10.1093/carcin/17.4.761
CR  - [27] Jayaprakasam B, Vanisree M, Zhang Y, Dewitt DL, Nair MG. Impact of alkyl esters of caffeic and ferulic acids on tumor cell proliferation, cyclooxygenase enzyme, and lipid peroxidation. J Agric Food Chem. 2006; 54(15): 5375-5381. https://doi.org/10.1021/jf060899p
CR  - [28] Eroğlu C, Seçme M, Bağcı G, Dodurga Y. Assessment of the anticancer mechanism of ferulic acid via cell cycle and apoptotic pathways in human prostate cancer cell lines. Tumor Biol. 2015; 36(12): 9437-9446. https://doi.org/10.1007/s13277-015-3689-3
CR  - [29] Okur ME, Ayla Ş, Karadağ AE, Polat DÇ, Demirci S, Seçkin İ. Opuntia ficus indica fruits ameliorate cisplatin-induced nephrotoxicity in mice. Biol Pharm Bull. 2020; 43(5): 831-838. https://doi.org/10.1248/bpb.b19-01044
CR  - [30] Sahin Z, Biltekin SN, Yurttas L, Berk B, Özhan Y, Sipahi H, Gao ZG, Jacobson KA, Demirayak Ş. Novel cyanothiouracil and cyanothiocytosine derivatives as concentration-dependent selective inhibitors of U87MG glioblastomas: Adenosine receptor binding and potent PDE4 inhibition. Eur J Med Chem. 2021; 212: 113125. https://doi.org/10.1016/j.ejmech.2020.113125
UR  - https://dergipark.org.tr/en/pub/jrespharm/issue//1690629
L1  - https://dergipark.org.tr/en/download/article-file/4833679
ER  -