POTENTIAL GENOTOXICITY AND HISTOPATHOLOGICAL ALTERATION EVALUATION OF HEPTEXÂ®
Keywords:Heptex, Genotoxicity, In vivo, In vitro, Chromosome aberrations, Micronucleus, Bacterial reverse mutation, Histopathology
Objective: The present study was performed in order to evaluate potential genotoxicity and the histopathological alteration of a traditional herbal prescription Heptex that used in the treatment of liver disease.
Methods: The genotoxicity were evaluated using the in vivo chromosome aberration and micronucleus assays in bone marrow cells of male and female Sprague-Dawley rats. In addition, in vitro chromosome aberration assay in Chinese hamster ovary (CHO) cells and bacterial reverse mutations assay in Salmonella typhimurium strains and Escherichia coli (WP2-uvrA/) with and without metabolic activation system (S9 mix) were performed. Histopathological study was conducted in liver, ovary and testis tissue of Sprague-Dawley rats.
Results: The genotoxicity assessment showed that Heptex did not significantly increase the number of chromosomal aberrations and frequencies of micro nucleated polychromatic erythrocytes in bone marrow cells of both male and female rats. In addition, there were no increases in the number of revertant colonies at any concentrations of Heptex used in the study. Heptex did not produce any structural aberration in CHO cells in the presence or absence of S9 mix. In addition, there were no histopathological changes induced by Heptex in rat liver, ovary and testis.
Conclusion: Based on abovementioned findings, we can conclude that Heptex is generally non-toxic and does not exhibit genotoxicity or histopathological alteration.
Samuel R, Kathriarachchi H, Hoffmann P, Barfuss M, Wurdack K, Davis C, et al. Molecular phylogenetics of Phyllanthaceae: evidence from plastid matK and nuclear PHYC sequences. Am J Bot 2005;92:132.
Ahmeda A. Isolation and characterization of gallic acid, Quercetin and methyl dehydrochebulate from phyllanthus niruri (Dukung Anak). Malaysian J Sci 2005;24:121-3.
Markom M, Hasan M, Daud W, Singh H, Jahim J. Extraction of hydrolysable tannins from Phyllanthus niruri L. In: Effects of solvents and extraction methods. Sep Purif Technol 2007;52:487-96.
Fang S, Rao Y, Tzeng Y. Anti-oxidant and inflammatory mediator's growth inhibitory effects of compounds isolated from Phyllanthus urinaria. J Ethnopharmacol 2008;116:333-40.
Murugaiyah V, Chan K. Determination of four lignans in Phyllanthus niruri L. by a simple high-performance liquid chromatography method with fluorescence detection. J Chromatogr A 2007;1154(1-2):198-204.
Chularojmontri L, Wattanapitayakul S, Herunsalee A, Charuchongkolwongse S, Niumsakul S, Srichairat S. Antioxidative and cardioprotective effects of Phyllanthus urinaria L. on doxorubicin-induced cardiotoxicity. Biol Pharm Bull 2005;28:1165-71.
Harish R, Shivanandappa T. Antioxidant activity and hepatoprotective potential of phyllanthus niruri. Food Chem 2006;95:180â€“5.
Ambali S, Adeniyi S, Makinde A, Shittu M, Yaqub L. Methanol extract of Phyllanthus niruri attenuates chlorpyrifos-evoked erythrocyte fragility and lipoperoxidative changes in Wister rats. Arch Appl Sci Res 2010;2:191-8.
Krithika R, Mohankumar R, Verma R, Shrivastav P, Mohamad I, Gunasekaran P, et al. Isolation, characterization and antioxidative effect of phyllanthin against CCl4-induced toxicity in HepG2 cell line. Chem-Biol Interact 2009;181:351-8.
Calixto JB, Santos AR, Filho VC, Yunes RA. A review of the plants of the genus Phyllanthus: their chemistry, pharmacology, and therapeutic potential. Med Res Rev 1998;18:225-58.
Hiraganahalli BD, Chinampudur VC, Dethe S, Mundkinajeddu D, Pandre MK, Balachandran J, et al. Hepatoprotective and antioxidant activity of standardized herbal extracts. Pharmacogn Mag 2012;8:116-23.
Pramyothin P, Ngamtin C, Poungshompoo S, Chaichantipyuth C. Hepatoprotective activity of Phyllanthus amarus Schum. Thonn. extract in ethanol treated rats: in vitro and in vivo studies. J Ethnopharmacol 2007;114:169-73.
Khatoon S, Rai V, Rawat AK, Mehrotra S. Comparative pharmacognostic studies of three Phyllanthus species. J Ethnopharmacol 2006;104:79-86.
Asha VV, Akhila S, Wills PJ, Subramoniam A. Further studies on the antihepatotoxic activity of Phyllanthus maderaspatensis Linn. J Ethnopharmacol 2004;92:67-70.
Tatiya AU, Surana SJ, Sutar MP, Gamit NH. Hepatoprotective effect of poly herbal formulation against various hepatotoxic agents in rats. Pharmacogn Res 2012;4:50-6.
Sabir SM, Rocha JBT. Water-extractable phytochemicals from Phyllanthus niruri exhibit distinct in vitro antioxidant and in vivo hepatoprotective activity against paracetamol-induced liver damage in mice. Food Chem 2008;111:845-51.
Khanna AK, Rizvi F, Chander R. Lipid lowering activity of Phyllanthus niruri in hyperlipemic rats. J Ethnopharmacol 2002;82:19-22.
Manjrekar AP, Jisha V, Bag PP, Adhikary B, Pai MM, Hegde A, et al. Effect of Phyllanthus niruri Linn. treatment on liver, kidney and testes in CCl4 induced hepatotoxic rats. Indian J Exp Biol 2008;46:514-20.
Comelli MC, Mengs U, Schneider C. Toward the defi nition of the mechanism of action of silymarin: activities related to cellular protection from toxic damage induced by chemo-therapy. Integr Cancer Ther 2007;6:120-9.
Kaleeswaran S, Sriram P, Vijay kumar C, Mathuram LN. Genotoxic and antigenotoxic activity of silymarin by an in vivo bone marrow micronucleus assay and comet assay. Iran J Toxicol 2009;2(3):203-10.
Kidd PM. Altern. Bioavailability and activity of phytosome complexes from botanical polyphenols: the silymarin, curcumin, green tea, and grape seed extracts. Med Rev 2009;14:226-46.
Ladas EJ, Kelly KM. Milk thistle: is there a role for its use as an adjunct therapy in patients with cancer. J Altern Complement Med 2003;9:411-6.
Post-White J, Ladas EJ, Kelly KM. Advances in the use of milk thistle (Silybum marianum). Integr Cancer Ther 2007;6(2):104-9.
Saller R, Brignoli R, Melzer J, Meier R. An updated systematic review with meta-analysis for the clinical evidence of silymarin. Forsch Komplement Med 2008;15:9-20.
Calixto JB. Efficacy, safety, quality control, marketing and regulatory guidelines for herbal medicines (phytotherapeutic agents). Braz J Med Biol Res 2000;33:179-89.
Yosida TH, Amano K. Autosomal polymorphism in laboratory bred and wild Norway rats, Rattus norvegius, found in Misima. Chromosoma 1965;16:658-67.
Nwanjo HU. Studies on the effect of aqueous extract of Phyllanthus niruri on plasma glucose level and some hepatospecific markers in diabetic Wistar rats. Internet J Lab Med 2007;2(2):1-18.
Asare GA, Addo P, Bugyei K, Gyan B, Adjei S, Otu-Nyarko LS, et al. Acute toxicity studies of aqueous leaf extract of Phyllanthus niruri. Interdiscip Toxicol 2011;4(4):206-10.
Razak MFA, Aidoo KE, Candlish AGG. Mutagenic and cytotoxic properties of three herbal plants from Southeast Asia. Tropic Biomed 2007;24(2):49-59.
Valerio LG, Gonzales GF. Toxicological aspects of the south american herbs Cat's Claw (Uncariatomentosa) and Maca (Lepidi-ummeyenii): a critical synopsis. Toxicol Rev 2005;24(1):11-35.
Wanyoike GN, Chhabra SC, Langâ€™at-Thoruwa CC, Omar SA. Brine shrimp toxicity and antiplasmodial activity of five Kenyan medicinal plants. J Ethnopharmacol 2004;90(1):129-33.
Gatehouse D, Haworth S, Cebula T, Gocke E, Kier L, Matsushima T, et al. Report from the working group on bacterial mutation assays: international workshop on standardization of genotoxicity test procedures. Mutat Res 1994;312:217-33.
Asare GA, Bugyei K, Sittie A, Yahaya ES, Gyan B, Adjei S, et al. Genotoxicity, cytotoxicity and toxicological evaluation of whole plant extracts of the medicinal plant Phyllanthus niruri (Phyllanthaceae). Genet Mol Res 2012;11(1):100-11.
De Queiroz FM, Matias KWO, Da Cunha MMF, Schwarz A. Evaluation of (anti)genotoxic activities of Phyllanthus niruri L. in rat bone marrow using the micronucleus test. Braz J Pharm Sci 2013;49(1):135-8.
Tona L, Mesia K, Ngimbi NP, Chrimwami B, Okond A, Cimanga K, et al. In vivo antimalarial activity of Cassia occidentalis, Morinda morindoides and Phyllanthus niruri. Ann Trop Med Parasitol 2001;95:47-57.
Makoshi MS, Adanyeguh IM, Nwatu LI. Hepatoprotective effect of Phyllanthus niruri aqueous extract in acetaminophen sub-acutec exposure rabbits. J Vet Med Anim Health 2013;5(1):8-15.
Gami B, Kothari IL. Antioxidant and antimicrobial activity of in vivo and in vitro grown plants of phyllanthus niruri L. Int J Pharm Biol Sci 2011;2(2):78-89.
Siddique YH, Ara G, Beg T, Afzal M. Protective role of nordihydroguaiaretic acid (NDGA) against the genotoxic damage induced by ethynodiol diacetate in human lymphocytes in vitro. J Environ Biol 2007;28:279-82.
Dixit N, Baboota S, Kohli K, Ahmad S, Ali J. Silymarin: a review of pharmacological aspects and bioavailability enhancement approaches. Indian J Pharmacol 2007;39:172-9.
Varzi HN, Esmailzadeh S, Morovvati H, Avizeh R, Shahriari A, Givi ME. Effect of silymarin and vitamin E on gentamicin induced nephrotoxicity in dogs. J Vet Pharmacol Ther 2007;30(5):477-81.
Toklu HZ, Tunali Akbay T, Velioglu-Ogunc A, Ercan F, Gedik N, Keyer-Uysal M, et al. Silymarin, the antioxidant component of Silybum marianum, prevents sepsis-induced acute lung and brain injury. J Surg Res 2008;145(2):214-22.
El-Kamary SS, Shardell MD, Abdel-Hamid M, Ismail S, El-Ateek M, Metwally M, et al. A Randomized Controlled Trial to Assess the Safety and Efficacy of Silymarin on Symptoms, Signs and Biomarkers of Acute Hepatitis. Phytomedicine 2009;16(5):391â€“400.
Shaker E, Mahmoud H, Mnaa S. Silymarin, the antioxidant component and Silybum marianum extracts prevent liver damage. Food Chem Toxicol 2010;48(3):803-6.