Main Article Content
The leaves of Perilla frutescens (PF) (Nga-Kee-Mon) have been widely used as a functional food and a traditional medicine for various therapeutic applications. This plant possesses many biological activities such as antioxidant, antibacterial, anti-inflammatory, and anticancer properties. However, safety information, especially toxicity studies are very limited. This study aimed to investigate the genotoxic effect of PF leaf extract on fibroblast V79 cell line using micronucleus (MN) test. The cytokinesis-block proliferation index (CBPI), percent cytostasis and MN frequencies were determined in V79 cells treated with the extract (100-250 µg/mL). The results showed that in the test condition without S9 enzyme fraction, the extract at the highest concentration (250 µg/mL) exhibited a higher cytotoxic effect than that with S9, as shown in percent cytostasis of the cells (57.47% and 34.00%, respectively). However, PF extract at all concentrations, with or without S9, did not significantly increase the MN frequency of the cells when compared to the negative control. The results suggested that PF leaf extract had neither direct nor indirect (metabolite form) genotoxic effect on V79 cells, thus enabling its use as a safe food supplement and medicine. Nevertheless, subchronic and chronic toxicity studies in animals should be further performed to provide more information regarding its long-term safety.
Upon acceptance of an article, the Pharmacological and Therapeutic Society of Thailand will have exclusive right to publish and distribute the article in all forms and media and grant rights to others. Authors have rights to use and share their own published articles.
Kongkeaw S, Riebroy S, Chaijan M. Comparative studies on chemical composition, phenolic compounds and antioxidant activities of brown and white perilla (Perilla frutescens) seeds. Chiang Mai J Sci. 2015;42(4):896-906.
Igarashi M, Miyazaki Y. A review on bioactivities of perilla: progress in research on the functions of perilla as medicine and food. Evid Based Complement Alternat Med. 2013;2013:925342.
Chen YP. Application and prescriptions of perilla in traditional Chinese medicine. In: Kosuna K, Haga M, Yu HC, editors. Perilla: The genus Perilla. Amsterdam: Harwood Academic Publishers; 1997. p. 37-45.
Meng L, Lozano Y, Bombarda I, Gaydou EM, Li B. Polyphenol extraction from eight Perilla frutescens cultivars. C R Chim. 2009 May;12(5):602-11.
Ahmed HM. Ethnomedicinal, phytochemical and pharmacological investigations of Perilla frutescens (L.) Britt. Molecules. 2018 Dec 28;24(1):102.
Saita E, Kishimoto Y, Tani M, Iizuka M, Toyozaki M, Sugihara N, et al. Antioxidant activities of Perilla frutescens against low-density lipoprotein oxidation in vitro and in human subjects. J Oleo Sci. 2012 Mar;61(3):113-20.
Izumi Y, Matsumura A, Wakita S, Akagi K, Fukuda H, Kume T, et al. Isolation, identification, and biological evaluation of Nrf2-ARE activator from the leaves of green perilla (Perilla frutescens var. crispa f. viridis). Free Radic Biol Med. 2012 Aug;53(4):669-79.
Qunqun G, Guicai DU, Ronggui LI. Antibacterial activity of Perilla frutescens leaf essential oil. Sci Technol Food Ind. 2003;9:25-27.
Huang BP, Lin CH, Chen YC, Kao SH. Anti-inflammatory effects of Perilla frutescens leaf extract on lipopolysaccharide-stimulated RAW264.7 cells. Mol Med Rep. 2014 Aug;10(2):1077-83.
Omer EA, Khattab ME, Ibrahim ME. First cultivation trial of Perilla frutescens L. in Egypt. Flavour Fragr J. 1998 Jul;13(4):221-5.
Banno N, Akihisa T, Tokuda H, Yasukawa K, Higashihara H, Ukiya M, et al. Triterpene acids from the leaves of Perilla frutescens and their anti-inflammatory and antitumor-promoting effects. Biosci Biotechnol Biochem. 2004 Jan;68(1): 85-90.
Lin CS, Kuo CL, Wang JP, Cheng JS, Huang ZW, Chen CF. Growth inhibitory and apoptosis inducing effect of Perilla frutescens extract on human hepatoma HepG2 cells. J Ethnopharmacol. 2007 Jul;112(3):557-67.
Pintha K, Tantipaiboonwong P, Yodkeeree S, Chaiwangyen W, Chumphukam O, Khantamat O, et al. Thai perilla (Perilla frutescens) leaf extract inhibits human breast cancer invasion and migration. Maejo Int J Sci Technol. 2018 May;12(2):112-23.
Khanaree C, Pintha K, Tantipaiboonwong P, Suttajit M, Chewonarin T. The effect of Perilla frutescens leaf on 1, 2‐dimethylhydrazine‐induced initiation of colon carcinogenesis in rats. J Food Biochem. 2018:e12493.
Kangwan N, Pintha K, Lekawanvijit S, Suttajit M. Rosmarinic acid enriched fraction from Perilla frutescens leaves strongly protects indomethacin-induced gastric ulcer in rats. Biomed Res Int. 2019 Mar 4;2019:9514703.
Fenech M. The micronucleus assay determination of chromosomal level DNA damage. Methods Mol Biol. 2008;410:185-216.
Ren N, Atyah M, Chen WY, Zhou CH. The various aspects of genetic and epigenetic toxicology: testing methods and clinical applications. J Transl Med. 2017 May 22;15(1):110.
Luzhna L, Kathiria P, Kovalchuk O. Micronuclei in genotoxicity assessment: from genetics to epigenetics and beyond. Front Genet. 2013 Jul 11;4:131.
Fenech M, Morley AA. Cytokinesis-block micronucleus method in human lymphocytes: effect of in vivo ageing and low dose X-irradiation. Mutat Res. 1986 Jul;161(2):193-8.
Organization of Economic Co-operation and Development. OECD guideline for testing of chemicals. Test No. 487: In vitro mammalian cell micronucleus test. [internet]. Paris: OECD publishing; 2016 [update 2016; cited 2019 Dec 20]. Available from: https://www.oecd.org/chemicalsafety/test-no-487-in-vitro-mammalian-cell-micronucleus-test-9789264264861-en.htm
Thomas P, Umegaki K, Fenech M. Nucleoplasmic bridges are a sensitive measure of chromosome rearrangement in the cytokinesis-block micronucleus assay. Mutagenesis. 2003 Mar;18(2):187-94.
Countryman PI, Heddle JA. The production of micronuclei from chromosome aberrations in irradiated cultures of human lymphocytes. Mutat Res. 1976 Dec; 41(2-3):321-32.
Himakoun L, Tuchinda P, Puchadapirom P, Tammasakchai R, Leardkamolkarn V. Evaluation of gonotoxic and anti-mutagenic properties of cleistanthin A and cleistanthoside A tetraacetate. Asian Pacific J Cancer Prev. 2011 Jan;12(12): 3271-5.
Bryce SM, Avlasevich SL, Bemis JC, Lukamowicz M, Elhajouji A, Goethem FV, et al. Interlaboratory evaluation of a flow cytometric, high content in vitro micronucleus assay. Mutat Res. 2008 Feb 29;650(2):181-95.
Krause AW, Carley WW, Webb WW. Fluorescent erythrosine B is preferable to trypan blue as a vital exclusion dye for mammalian cells in monolayer culture. J Histochem Cytochem. 1984 Oct;32(10):1084-90.
Weisenthal LM, Dill PL, Kurnick NB, Lippman ME. Comparison of dye exclusion assays with a clonogenic assay in the determination of drug-induced cytotoxicity. Cancer Res. 1983 Jan;43(1):258-64.
Kim SI, Kim HJ, Lee HKL, Hong D, Lim H, Cho K, et al. Application of non-hazardous vital dye for cell counting with automated cell counters. Anal Biochem. 2016 Jan 1;492:8-12.
Son YO, Kim J, Kim JC, Chung Y, Chung GH, Lee JC. Ripe fruits of Solanim nigrum L. inhibits cell growth and induces apoptosis in MCF-7 cells. Food Chem Toxicol. 2003 Oct;41(10):1421-8.
Lee JC, Lee KY, Son YO, Choi KC, Kim J, Truong TT, et al. Plant-originated glycoprotein, G-120, inhibits the growth of MCF-7 cells and induces their apoptosis. Food Chem Toxicol. 2005 Jun;43(6):961-8.
Yamamoto M, Motegi A, Seki J, Miyamae Y. The optimized conditions for the in vitro micronucleus (MN) test procedures using chamber slides. Environ Mutagen Res. 2005 Jul;27(3):145-51.
Lorge E, Hayashi M, Albertini S, Kirkland D. Comparison of different methods for an accurate assessment of cytotoxicity in the in vitro micronucleus test. I. theoretical aspects. Mutat Res. 2008 Aug-Sep;655(1-2):1-3.
Organization of Economic Co-operation and Development. OECD guideline for testing of chemicals. Proposal for updating test guideline 473: In vitro mammalian chromosome aberration test. [Internet]. Paris: OECD publishing; 2012 [update 2012; cited 2020 Jun 23]. Available from: http://www.oecd.org/ chemicalsafety/ testing/50108781.pdf.