Mutagenic and antimutagenic evaluation of Asparagus laricinus Burch., Senecio asperulus DC., and Gunnera perpensa L. to hepatic cells




neutral red uptake, VITOTOX, Alkaline Comet assay, VITOTOX , Alkaline Comet assay, DNA mutation toxicity


Introduction. The use of traditional medicinal plant concoctions to cure or treat different diseases daily in African folk medicine. However, the effects of most medicinal plants on human health or genetic material remain unknown. This study thus aimed to evaluate the mutagenic and antimutagenic potentials of Asparagus laricinus Burch. cladodes, Senecio asperulus DC., and Gunnera perpensa L. roots extract in vitro.

Material and methods. Neutral red uptake assay, alkaline comet assay, and the VITOTOX test was used with plant extract dilutions of 4, 20, 50, and 100 µg/ml, respectively, on hepatic (C3A) cells and Salmonella Typhimurium TA104 strains. Ethyl methane-sulfonate and 4-nitroquinoline oxide were used as positive controls for the comet and VITOTOX assays, respectively.

Results.  In vitro cytotoxicity and genotoxicity were not observed from all tested extracts, except for the two dichloromethane (DCM) extracts of S. asperulus and G. perpensa, which appeared to be cytotoxic with S9 metabolic activation, but not genotoxic or mutagenic. From the VITOTOX test results, none of the extracts appeared to have antimutagenic properties after treating S. Typhimurium strains with a known mutagen.

Conclusions. These results confirm that previously reported anticarcinogenic properties of A. laricinus, S. asperulus, and G. perpensa did not result from the protective mechanism against genotoxicity but from other ones. Moreover, the negative mutagenic and cytotoxic activities of the tested plants highlighted the safe use of these medicinal plants in vitro. Therefore, S. asperulus and G. perpensa DCM extracts require further investigation for their possible in vivo cytotoxic effects on humans.


Download data is not yet available.


Abd Razak, M, F., K.E. Aidoo and A.G. Candlish, 2007. Mutagenic and cytotoxic properties of three herbal plants from Southeast Asia. Trop Biomed, 24(2): 49-59.

Roy, A., T. Attre and N. Bharadvaja, 2017. Anticancer agent from medicinal plants: a review. In book: New aspects in medicinal plants and pharmacognosy, 1st edition.;1(3): 54-73.

Verschaeve, L., H. Edziri, R. Anthonissen, D. Boujnah, F. Skhiri and H Chehab, 2017. In vitro toxicity and genotoxic activity of aqueous leaf extracts from four varieties of Olea europea (L). Pharmacog Magazine, 13(S1): S63-S80.

Verschaeve, L., J. Van Gompel, L. Thilemans, L. Regniers, P. Vanparys and D. Van der Lelie, 1999. VITOTOX® bacterial genotoxicity and toxicity test for the rapid screening of chemicals. Environ and Mol Mutagen, 33(3): 240-8.;2-R

Griffiths, A.J., J.H. Miller, D.T. Suzuki, R.C. Lewontin and W.M. Gelbart, 2000. Quantifying heritability. In Freeman WH, ed. An Introduction to Genetic Analysis, 7th edition..

Bouguellid, G., C. Russo, M. Lavorgna, C. Piscitelli, K. Ayouni, E. Wilson, H.K. Kim, R. Verpoorte, Y.H. Choi, D. Kilani-Atmani, D. Atmani, M. Isidori, 2020. Antimutagenic, antigenotoxic and antiproliferative activities of Fraxinus angustifolia Vahl. leaves and stem bark extracts and their phytochemical composition. PLoS One, 16;15(4):e0230690.

Mustapha, A.A, 2014. Medicinal plants with possible anti-HIV activities: A review. Int J Med Plants, 106: 439-53

Mashele, S.S. and N. Kolesnikova, 2010. In vitro anticancer screening of Asparagus laricinus extracts. Pharmacologyonline, 2(1): 246–252.

Mfengwana, P.H. and S.S Mashele, 2019. Medicinal Properties of Selected Asparagus Species: A Review. In: Roa V, Mans D, Roa L, eds. Phytochemicals in Human Health. London. Intechopen.

Fuku, S., A.M. Al-Azzawi, I.T. Madamombe-Manduna and S. Mashele, 2013. Phytochemistry and free radical scavenging activity of Asparagus laricinus. Int J Pharmacol.9(5): 312-17.

Ntsoelinyane, P.H. and S.S Mashele, 2014. Phytochemical screening, antibacterial and antioxidant activities of Asparagus laricinus leaf and stem extracts. Bangladesh J Pharmacol, 9: 10-4.

Mfengwana, P.H., S.S. Mashele and I.T. Manduna, 2019. Cytotoxicity and cell cycle analysis of Asparagus laricinus Burch. and Senecio asperulus DC. on breast and prostate cancer cell lines. Heliyon, 5(5):e01666.

Mugomeri, E., P. Chatanga, S. Hlapisi, L. Rahlao, 2014. Phytochemical characterization of selected herbal products in Lesotho. Lesotho Med Asso J, 12: 38–47.

Moteetee, A. and B. Van Wyk, 2011. The medical ethnobotany of Lesotho: a review. Bothalia, 41(1): 209-228.

Kose, L.S., A. Moteetee, S. Van Vuuren, 2015. Ethnobotanical survey of medicinal plants used in the Maseru district of Lesotho. J Ethnopharmacol, 170: 184–200.

Quattrocchi, U, 2016. CRC World Dictionary of Medicinal and Poisonous Plants. Common Names, Scientific Names, Eponyms, Synonyms, and Etymology. Boca Raton: Taylor & Francis Group.

Mammo, F., V. Mohanlall and F. Shode, 2017. Gunnera perpensa L.: a multi-use ethnomedicinal plant species in South Africa. Afr J Sci Technol Innov Dev. DOI: 10.1080/20421338.2016.1269458

Maroyi, A, 2016. From Traditional Usage to Pharmacological Evidence: Systematic Review of Gunnera perpensa L. J Evid Based Complement Altern Med, 17: 14-25.

Simelane, M., O.A. Lawal, T.G. Djarova, C.T. Musabayane, M. Singh, and A.R. Opoku, 2012. Lactogenic activity of rats stimulated by Gunnera perpensa L. (Gunneraceae) from South Africa. Afr J Tradit, Complement Altern Med, 9(4): 561–573.

Chigor, C.B, 2014. Development of conservation methods for Gunnera perpensa L.: an overexploited medicinal plant in the Eastern Cape, South Africa. PhD Thesis, University of Fort Hare..

Khan, F., X.K. Peter, R.M. Mackenzie, L. Katsoulis, R. Gehring, O.Q. Munro, R. van Heerden and S.E. Drewes, 2004. “Venusol From Gunnera perpensa: Structural and Activity Studies.” Phytochemistry, 65(8): 1117–21.

Ndhlala, A.R., J.F. Finnie and J. Van Staden, 2011. Plant composition, pharmacological properties and mutagenic evaluation of a commercial Zulu herbal mixture: Imbiza ephuzwato. J Ethnopharmacol, 133(2): 663-74.

Efferth, T. and B. Kaina, 2011. Toxicities by herbal medicines with emphasis to traditional Chinese medicine. Curr Drug Metab, 12(10): 989-96.

Cardoso, C.R., I.M. de Syllos Cólus, C.C. Bernardi, M. Sannomiya, W. Vilegas, E.A. Varanda, 2006. Mutagenic activity promoted by amentoflavone and methanolic extract of Byrsonima crassa Niedenzu. Toxicol, 225(1): 55-63.

Déciga-Campos, M., I. Rivero-Cruz, M. Arriaga-Alba, G. Castañeda-Corral, G.E. Angeles-López, A. Navarrete. And R. Mata, 2007. Acute toxicity and mutagenic activity of Mexican plants used in traditional medicine. J Ethnopharmacol, 110(2): 334-42.

Repetto, G., A. Del Peso. and J.L. Zurita, 2008. Neutral red uptake assay for the estimation of cell viability/cytotoxicity. Nat Protoc, 3(7): 1125-31.

Olive, P.L. and J.P. Banáth, 2006. The comet assay: a method to measure DNA damage in individual cells. Nat Protoc, 1: 23–29

Verschaeve, L., 2005. The VITOTOX® genotoxicity test. In Pandalai SG, ed. Recent research developments in applied microbiology and biotechnology. India: Research Signpost;.pp. 33-49.

Collins, A.R., 2011. The use of bacterial repair endonucleases in the comet assay. In Gautier JC, ed. Drug Safety Evaluation. Methods in Molecular Biology, Vol 691.New Jersey: Humana Press; pp. 137-47.

Makhuvele, R., K. Foubert, S. Apers, L. Pieters, L. Verschaeve and E. Elgorashi, 2018. Antimutagenic constituents from Monanthotaxis caffra (Sond.) Verdc. J Pharm Pharmacol, 70(7): 976-84.

Vaghasiya, Y., R. Dave and S. Chanda, 2011. Phytochemical analysis of some medicinal plants from western region of India. Res J Medicinal Plant, 5(5): 567-76.

Biran, A., H.B. Yoav, S. Yagur-Kroll, R. Pedahzur, S. Buchinger, Y. Shacham-Diamand, G. Reifferscheid and S. Belkin, 2011. Microbial genotoxicity bioreporters based on sulA activation. Anal Bioanal Chem, 400(9): 3013-24.

Słoczyńska, K., B. Powroźnik, E. Pękala and A.M. Waszkielewicz, 2014. Antimutagenic compounds and their possible mechanisms of action. J Appl Genet, 55(2): 273-85.

Azqueta, A. and A.R. Collins, 2013. The essential comet assay: a comprehensive guide to measuring DNA damage and repair. Arch Toxicol, 87(6): 949-68.

Chang, J.B., M.F. Wu, H.F. Lu, J, Chou, M.K. Au, N.C. Liao, C.H. Chang, Y.P. Huang, C.T. Wu, and J.G. Chung, 2013. Toxicological evaluation of Antrodia cinnamomea in BALB/c mice. In Vivo, 27(6): 739-45.

Mashele, S.S. and S. Fuku, 2011. Evaluation of the antimutagenic and mutagenic properties of Asparagus laricinus. J Med Technol, 2: 33-36.

Mfengwana, P, 2019. Evaluation of Pharmacological Properties of Traditional Medicinal Plants Used For The Treatment Of Cancer By South African And Lesotho Communities.

Mokgawa, S.D, 2016. Toxicology of Asparagus laricinus in rats.

Edziri, H., M. Mastouri, A. Mahjoub, R. Anthonissen, B. Mertens, S. Cammaerts, L. Gevaert and L. Verschaeve, 2011. Toxic and mutagenic properties of extracts from Tunisian traditional medicinal plants investigated by the neutral red uptake, VITOTOX and alkaline comet assays. S Afr J Bot, 77: 703–710.

Gautam, S., S. Saxena and S. Kumar, 2016. Fruits and vegetables as dietary sources of antimutagens. J Food Chem Nanotechnol, 2(3): 97-114.

El-Sayed, W.M. and W.A. Hussin, 2013. Antimutagenic and antioxidant activity of novel 4-substituted phenyl-2, 2′-bichalcophenes and aza-analogs. Drug Des Dev Ther, 7: 73-81.

Ferguson, L.R. and M. Philpott, 2008. Nutrition and mutagenesis. Annu. Rev. Nutr, 28:313-29.

Fronza, G., P. Campomenosi, R. lannone, and A. Abbondandolo, 1992. The 4-nitroquinoline 1-oxide mutational spectrum in single-stranded DNA is characterized by guanine to pyrimidine transversions. Nucleic Acids Res, 20(6): 1283-7.

Brookes, K.B. and M.F. Dutton, 2007. Bioactive Components of the Uteroactive Medicinal Plant, Gunnera Perpensa (or Ugobo). S Afr J Sci, 103 (5): 187–189.

How to Cite

Mfengwana P-M-AH. Mutagenic and antimutagenic evaluation of Asparagus laricinus Burch., Senecio asperulus DC., and Gunnera perpensa L. to hepatic cells. JMS [Internet]. 2022 Dec. 27 [cited 2023 Feb. 4];91(4):e745. Available from:



Original Papers
Received 2022-09-25
Accepted 2022-10-23
Published 2022-12-27