Evaluate the Effect of Ethanol Fraction of Iraqi Hibiscus tiliaceus L. Leaves Extract on Mitotic Index and Micronucleus Frequency in Comparison with Metformin in Male Rats
DOI:
https://doi.org/10.31351/vol34iss4pp141-149Keywords:
Hibiscus tiliaceus L, Mitotic index, Micronucleus appearance, Methotrexate, MetforminAbstract
Hibiscus tiliaceus L. is a member of the Malvaceae family. It has been documented to exhibit antibacterial, antioxidant, anti-inflammatory, expectorant, anthelmintic, and anticancer properties. The objective of this study was to assess the impact of ethanol leaf extract from Hibiscus tiliaceus L. on methotrexate-induced chromosomal aberrations, specifically the Mitotic Index and Micronucleus Frequency, in spleen and bone marrow cells.
The comparison was made with metformin. The study involves the categorization of the animals into five groups, with each group consisting of six rats. The rats are then subjected to the following treatments: Group I (negative control) consisted of six rats that were administered a daily oral dose of 1ml/kg/day of distilled water for seven consecutive days. Group II (positive control): Six rats were administered a solitary dose of 20mg/kg of methotrexate on the initial day. Group III consisted of six rats who were administered a daily oral dose of 100 mg/kg of metformin for a period of seven consecutive days. Group IV consisted of six rats who were administered a daily oral dose of 250 mg/kg of the ethanol fraction for seven consecutive days. In Group V, six rats were administered a daily oral dose of 500 mg/kg of the ethanol fraction for seven consecutive days. The results indicate that the ethanol extract of Hibiscus tiliaceus leaves, at both doses, significantly reduced the mitotic index in bone marrow cells and spleen cells compared to the negative control group. Additionally, it significantly increased the appearance of micronuclei in bone marrow cells. In conclusion, the ethanol extract of Hibiscus tiliaceus leaves exhibited a strong effect on the mitotic index and micronucleus appearance in bone marrow and spleen cells of rats.
How to Cite
Publication Dates
Received
Revised
Accepted
Published Online First
References
Wang Z, Qi F, Cui Y, Zhao L, Sun X, Tang W, Cai P. An update on Chinese herbal medicines as adjuvant treatment of anticancer therapeutics. Bioscience trends. 2018 Jun 30;12(3):220-39.
Bedi O, Bijjem KR, Kumar P, Gauttam V. Herbal Induced Hepatoprotection and Hepatotoxicity: A Critical Review. Indian journal of physiology and pharmacology. 2016 Jan 1;60(1):6-21.
Gupta R, Kannan GM, Sharma M, Flora SJ. Therapeutic effects of Moringa oleifera on arsenic-induced toxicity in rats. Environmental toxicology and pharmacology. 2005 Nov 1;20(3):456-64.
Ramproshad S, Afroz T, Mondal B, Haque A, Ara S, Khan R, Ahmed S. Antioxidant and antimicrobial activities of leaves of medicinal plants Hibiscus tiliaceus L. Pharmacologyonline. 2012 Dec 30;3:82-7.
Samsudin MS, Andriani Y, Sarjono PR, Syamsumir DF. Study on Hibiscus tiliaceus leaves as antibacterial and antioxidant agents. Alotrop. 2019 Dec 19;3(2).
Vijay T, Rajendra B. Phytochemical screening and anthelmintic activity of wood and leaves of Hibiscus tiliaceus Linn. World Journal of Pharmacy and Pharmaceutical Sciences. 2014;3(10):880-9.
Mikkelsen TS, Thorn CF, Yang JJ, Ulrich CM, French D, Zaza G, Dunnenberger HM, Marsh S, McLeod HL, Giacomini K, Becker ML. PharmGKB summary: methotrexate pathway. Pharmacogenetics and genomics. 2011 Oct 1;21(10):679-86.
Homady MH, Kadhim HA, Al-Kelaby KK, Aziz DZ, Kadhim NJ. Cytotoxic activity of compounded anthracycline against rhabdomyosarcoma cancer cell line.Plant Archives, (2018) 18(1), 941-946.
Xin N, Fen Z, Li C, Yan X, Runming J. Intracranial hemorrhage following oral low-dose methotrexate after multiple toxicities caused by high-dose methotrexate in childhood acute lymphoblastic leukemia. Frontiers in Pharmacology. 2019 Sep 19;10:1072.
Nisbet JC, Sturtevant JM, Prins JB. Metformin and serious adverse effects. The Medical Journal of Australia. 2004 Jul 5;181(1):56.
Rababa'h AM, Alzoubi KH, Khabour OF, Ababneh M. Ameliorative effect of metformin on methotrexate-induced genotoxicity: An in vitro study in human cultured lymphocytes. Biomedical Reports. 2021 Jul 1;15(1):1-7.
Luong RH, Baer KE, Craft DM, Ettinger SN, Scase TJ, Bergman PJ. Prognostic significance of intratumoral microvessel density in canine soft-tissue sarcomas. Veterinary pathology. 2006 Sep;43(5):622-31.
Tapia C, Kutzner H, Mentzel T, Savic S, Baumhoer D, Glatz K. Two mitosis-specific antibodies, MPM-2 and phospho-histone H3 (Ser28), allow rapid and precise determination of mitotic activity. The American journal of surgical pathology. 2006 Jan 1;30(1):83-9.
Beresford MJ, Wilson GD, Makris A. Measuring proliferation in breast cancer: practicalities and applications. Breast Cancer Research. 2006 Dec;8:1-1.
Mai X, Zhou F, Lin P, Lin S, Gao J, Ma Y, Fan R, Ting W, Huang C, Yin D, Kang Z. Metformin scavenges formaldehyde and attenuates formaldehyde‐induced bovine serum albumin crosslinking and cellular DNA damage. Environmental toxicology. 2020 Nov;35(11):1170-8.
Fenech M, Chang WP, Kirsch-Volders M, Holland N, Bonassi S, Zeiger E. HUMN project: detailed description of the scoring criteria for the cytokinesis-block micronucleus assay using isolated human lymphocyte cultures. Mutation Research/Genetic Toxicology and Environmental Mutagenesis. 2003 Jan 10;534(1-2):65-75.
Fenech M, Kirsch-Volders M, Natarajan AT, Surralles J, Crott JW, Parry J, Norppa H, Eastmond DA, Tucker JD, Thomas P. Molecular mechanisms of micronucleus, nucleoplasmic bridge and nuclear bud formation in mammalian and human cells. Mutagenesis. 2011 Jan 1;26(1):125-32.
Kasamoto S, Masumori S, Hayashi M. In vivo micronucleus assay in mouse bone marrow and peripheral blood. Genotoxicity Assessment: Methods and Protocols. 2013:179-89.
Mahire SP, Patel SN. Extraction of phytochemicals and study of its antimicrobial and antioxidant activity of Helicteres isora L. Clin Phytoscience. 2020;6(1):1–6.
Rawool PP, Parulekar BC. Phytochemical screening of Hibiscus tiliaceus by FTIR spectroscopic analysis. Int. J. Pharm. Biol. Sci. 2019;9(3):1308-19.
Mutlag SH, Hamad MN, Abbas IS, Ismael SH. The evaluation of ethyl acetate fraction of Cressa cretica effect on mitotic index and micronucleous Frequency in Mice. Int J Pharm Sci Rev Res. 2017;45(28):147-50.
Abd El-Rahim AH, Abd-Elmoneim OM, Hafiz NA. Assessment of antigenotoxic effect of nanoselenium and metformin on diabetic rats. Jordan J Biol Sci. 2017 Sep;10(3):159-65.
Narender KS, Kumar D, Kumar V. Antinociceptive and anti-inflammatory activity of Hibiscus tiliaceus leaves. International Journal of Pharmacognosy and Phytochemical Research. 2009;1(1):15-7.
Allen JW, Shuler CF, Mendes RW, Latt SA. A simplified technique for in vivo analysis of sister-chromatid exchanges using 5-bromodeoxyuridine tablets. Cytogenetic and Genome Research. 1977 May 2;18(4):231-7.
Hayashi M, Sofuni T, Ishidate Jr M. An application of acridine orange fluorescent staining to the micronucleus test. Mutation Research Letters. 1983 Jun 1;120(4):241-7.
Chan EW, Wong SK, Chan HT. A review on the phytochemistry and pharmacology of two Hibiscus species with spectacular flower colour change: H. tiliaceus and H. mutabilis. International Journal of Pharmacognosy and Phytochemical Research. 2016;8(7):1200-8.
Martin SA, McCarthy A, Barber LJ, Burgess DJ, Parry S, Lord CJ, Ashworth A. Methotrexate induces oxidative DNA damage and is selectively lethal to tumour cells with defects in the DNA mismatch repair gene MSH2. EMBO molecular medicine. 2009 Sep 28;1(6‐7):323-37.
El–Alfy NZ, Mahmoud MF, El-Ashry SR, Alqosaibi AI. Genotoxic effect of methotrexate on bone marrow chromosomes and DNA of male albino mice (Mus musculus). The Egyptian journal of hospital medicine. 2016 Jul 1;64(1):350-63.
Giordano A, Tommonaro G. Curcumin and cancer. Nutrients. 2019 Oct 5;11(10):2376.
Luo X, Hu R, Zheng Y, Liu S, Zhou Z. Metformin shows anti‐inflammatory effects in murine macrophages through Dicer/microribonucleic acid‐34a‐5p and microribonucleic acid‐125b‐5p. Journal of Diabetes Investigation. 2020 Jan;11(1):101-9.
Nna VU, Bakar AB, Lazin MR, Mohamed M. Antioxidant, anti-inflammatory and synergistic anti-hyperglycemic effects of Malaysian propolis and metformin in streptozotocin–induced diabetic rats. Food and chemical toxicology. 2018 Oct 1;120:305-20.
Yang X, Ding H, Qin Z, Zhang C, Qi S, Zhang H, Yang T, He Z, Yang K, Du E, Liu C. Metformin prevents renal stone formation through an antioxidant mechanism in vitro and in vivo. Oxidative medicine and cellular longevity. 2016;2016(1):4156075.
Kolivand S, Motevaseli E, Cheki M, Mahmoudzadeh A, Shirazi A, Fait V. The anti-apoptotic mechanism of metformin against apoptosis induced by ionizing radiation in human peripheral blood mononuclear cells. Klin Onkol. 2017 Jan 1;30(5):372-9.
Chukwunonso Obi B, Chinwuba Okoye T, Okpashi VE, Nonye Igwe C, Olisah Alumanah E. Comparative study of the antioxidant effects of metformin, glibenclamide, and repaglinide in alloxan‐induced diabetic rats. Journal of diabetes research. 2016;2016(1):1635361.
Tang G, Yang H, Chen J, Shi M, Ge L, Ge X, Zhu G. Metformin ameliorates sepsis-induced brain injury by inhibiting apoptosis, oxidative stress and neuroinflammation via the PI3K/Akt signaling pathway. Oncotarget. 2017 Nov 11;8(58):97977.
Cahova M, Palenickova E, Dankova H, Sticova E, Burian M, Drahota Z, Cervinkova Z, Kucera O, Gladkova C, Stopka P, Krizova J. Metformin prevents ischemia reperfusion-induced oxidative stress in the fatty liver by attenuation of reactive oxygen species formation. American Journal of Physiology-Gastrointestinal and Liver Physiology. 2015 Jul 15;309(2):G100-11.
Luzhna L, Kathiria P, Kovalchuk O. Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond. Frontiers in genetics. 2013 Jul 11;4:131.
Rosa RM, Moura DJ, Melecchi MI, dos Santos RS, Richter MF, Camarao EB, Henriques JA, de Paula Ramos AL, Saffi J. Protective effects of Hibiscus tiliaceus L. methanolic extract to V79 cells against cytotoxicity and genotoxicity induced by hydrogen peroxide and tert-butyl-hydroperoxide. Toxicology in vitro. 2007 Dec 1;21(8):1442-52.
Abdul-Azeez ZM, Mutlag SH. Possible Protective Anticancer Effect of Ethanol Fraction of Iraqi Hibiscus tiliaceus L. Leaves Extract on Diethylnitrosamine-induced Hepatocarcinogenesis in Male Rats. Iraqi Journal of Pharmaceutical Sciences (P-ISSN 1683-3597 E-ISSN 2521-3512). 2023 Nov 3;32(Suppl.):145-55.
Abdul-Azeez ZM, Mutlag SH. Possible protective anticancer effect of chloroform fraction of Iraqi Hibiscus tiliaceus L. leaves extract on diethylnitrosamine-induced hepatocarcinogenesis in male rats. Journal of Complementary and Integrative Medicine. 2024 Jan 19(0).
Abdul-Azeez ZA, Mutlag SH. Possible protective anticancer effect of ethyl acetate fraction of iraqi hibiscus tiliaceus l. Leaves extract on diethylnitrosamine-induced hepatocarcinogenesis in male rats. Farmacia. 2024 Jan 1;72(1).
Alcaraz M, Olivares A, Achel DG, García-Gamuz JA, Castillo J, Alcaraz-Saura M. Genoprotective effect of some flavonoids against genotoxic damage induced by X-rays in vivo: relationship between structure and activity. Antioxidants. 2021 Dec 30;11(1):94.
Umran MA, Aziz GM, Yaseen NY. The ability of polyphenols extracted from green tea Camellia sinensis in influence of genetic cytotoxicity of catechol on mice bone marrow cells (in vivo). Journal of Biotechnology Research Center. 2009 Jun 1;3(2):76-90.
Azqueta A, Collins A. Polyphenols and DNA damage: A mixed blessing. Nutrients. 2016 Dec 3;8(12):785.
Abdul-Awal SM, Nazmir S, Nasrin S, Nurunnabi TR, Uddin SJ. Evaluation of pharmacological activity of Hibiscus tiliaceus. Springerplus. 2016 Dec;5:1-6.
Mohammed SK, Mutlag SH. Potential anti-obesity effects of two-graded doses of Iraqi Hibiscus tiliaceus leaves extract, alone and in combination with orlistat, on high-fat diet-induced obesity in male rats. Journal of Medicine and Life. 2023 Sep;16(9):1338.
Bergman ME, Davis B, Phillips MA. Medically useful plant terpenoids: biosynthesis, occurrence, and mechanism of action. Molecules. 2019 Nov 1;24(21):3961.
Downloads
Published
Issue
Section
License
Copyright (c) 2025 Iraqi Journal of Pharmaceutical Sciences
This work is licensed under a Creative Commons Attribution 4.0 International License.
