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In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts

Yıl 2023, Cilt: 27 Sayı: 5, 2115 - 2123, 28.06.2025

Öz

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.

Kaynakça

  • [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
  • [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.
  • [3] Zafar HI, Badiaa L. Ethnopharmacology of the plants of genus Ajuga. Pakistan J Pharm Sci. 2009; 22(4): 425-462.
  • [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.
  • [5] Khare CP. Indian Medicinal Plants-An Illustrated Dictionary. 1st Indian Reprint, Springer (India). 2007; 28.
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
Yıl 2023, Cilt: 27 Sayı: 5, 2115 - 2123, 28.06.2025

Öz

Kaynakça

  • [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
  • [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.
  • [3] Zafar HI, Badiaa L. Ethnopharmacology of the plants of genus Ajuga. Pakistan J Pharm Sci. 2009; 22(4): 425-462.
  • [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.
  • [5] Khare CP. Indian Medicinal Plants-An Illustrated Dictionary. 1st Indian Reprint, Springer (India). 2007; 28.
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
  • [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
Toplam 30 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Farmakognozi
Bölüm Articles
Yazarlar

Ayşegül Çaşkurlu Öz 0000-0001-7277-920X

Şule Nur Karavuş 0000-0001-9265-8042

Sevde Nur Biltekin 0000-0003-1896-2729

Esra Zeynep Helvaci 0000-0003-4765-6026

Ayşe Esra Karadağ

Yayımlanma Tarihi 28 Haziran 2025
Yayımlandığı Sayı Yıl 2023 Cilt: 27 Sayı: 5

Kaynak Göster

APA Çaşkurlu Öz, A., Karavuş, Ş. N., Biltekin, S. N., Helvaci, E. Z., vd. (2025). In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts. Journal of Research in Pharmacy, 27(5), 2115-2123.
AMA Çaşkurlu Öz A, Karavuş ŞN, Biltekin SN, Helvaci EZ, Karadağ AE. In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts. J. Res. Pharm. Temmuz 2025;27(5):2115-2123.
Chicago Çaşkurlu Öz, Ayşegül, Şule Nur Karavuş, Sevde Nur Biltekin, Esra Zeynep Helvaci, ve Ayşe Esra Karadağ. “In Vitro Cytotoxicity Evaluation and Phytochemical Analysis of Ajuga Reptans L. Extracts”. Journal of Research in Pharmacy 27, sy. 5 (Temmuz 2025): 2115-23.
EndNote Çaşkurlu Öz A, Karavuş ŞN, Biltekin SN, Helvaci EZ, Karadağ AE (01 Temmuz 2025) In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts. Journal of Research in Pharmacy 27 5 2115–2123.
IEEE A. Çaşkurlu Öz, Ş. N. Karavuş, S. N. Biltekin, E. Z. Helvaci, ve A. E. Karadağ, “In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts”, J. Res. Pharm., c. 27, sy. 5, ss. 2115–2123, 2025.
ISNAD Çaşkurlu Öz, Ayşegül vd. “In Vitro Cytotoxicity Evaluation and Phytochemical Analysis of Ajuga Reptans L. Extracts”. Journal of Research in Pharmacy 27/5 (Temmuz 2025), 2115-2123.
JAMA Çaşkurlu Öz A, Karavuş ŞN, Biltekin SN, Helvaci EZ, Karadağ AE. In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts. J. Res. Pharm. 2025;27:2115–2123.
MLA Çaşkurlu Öz, Ayşegül vd. “In Vitro Cytotoxicity Evaluation and Phytochemical Analysis of Ajuga Reptans L. Extracts”. Journal of Research in Pharmacy, c. 27, sy. 5, 2025, ss. 2115-23.
Vancouver Çaşkurlu Öz A, Karavuş ŞN, Biltekin SN, Helvaci EZ, Karadağ AE. In vitro cytotoxicity evaluation and phytochemical analysis of Ajuga reptans L. extracts. J. Res. Pharm. 2025;27(5):2115-23.