Efek antigenotoksik ekstrak etanolik daun sirsak (Annona muricata Linn) terhadap frekuensi mikronukleus mukosa bukal tikus Sprague Dawley

Tyas Prihatiningsih; Tetiana Haniastuti; Dewi Agustina

doi:10.22146/majkedgiind.11794

Efek antigenotoksik ekstrak etanolik daun sirsak (Annona muricata Linn) terhadap frekuensi mikronukleus mukosa bukal tikus Sprague Dawley

https://doi.org/10.22146/majkedgiind.11794

Tyas Prihatiningsih^(1*), Tetiana Haniastuti⁽²⁾, Dewi Agustina⁽³⁾

(1) Post graduate programme, Dentistry faculty, Gadjah mada University
(2) Departemen Biologi Mulut, Fakultas Kedokteran Gigi, Universitas Gadjah Mada, Yogyakarta
(3) Departemen Ilmu Penyakit Mulut, Fakultas Kedokteran Gigi, Universitas Gadjah Mada, Yogyakarta
(*) Corresponding Author

Abstract

The effect of soursop leaves (Annona uricata linn) ethanolic extract on micronucleus frequency of buccal mucosa epithelium of Sprague dawley rats. Polycyclic aromatic hydrocarbons is one of the largest groups of carcinogen in environment. 7,12-Dimetillbez (α) antransena is a compound of PAH class that has genotoxic carcinogen potency. One of the most frequently applied genotoxicity tests is micronucleus test. Soursop is a plant that can grow well in Indonesia. Its leaves contain avonoid and acetogenin assumed to have potential chemopreventive and anticancer activities. The aim of this study was to assess the antigenotoxic effect soursop leaves ethanolic extraction the micronucleus frequency of DMBA-induced buccal mucosa of rat. This research was conducted on 24 male Sprague Dawley rats aged 5 weeks and divided into six groups. Carcinogenesis on the lingual dorsum of group I-III were induced by DMBA topically 3 times a week for 16 weeks, group II and III were not only induced by carcinogenesis, but also were given soursop leaves ethanolic extract of 100 and 200 mg/kg body weight for 18 weeks, group IV was given soursop leaves ethanolic extract 200 mg/kg body weight, group V was given DMSO 1% and group VI was given no treatment. After 18th week, buccal mucosa swab for micronucleus test was conducted and stained with Feulgen-Rossenbeck method. The number of micronucleus is calculated under a light microscope, data were analized using using one-way ANOVA followed by Tukey HSD. The result showed that the average of buccal micronucleus frequency of group II (13 ± 0.82) and group III (12 ± 0.96) were decrease signicantly (p<0,05) than group I (24 ± 1.71). From the experiment, it is concluded that the soursop leaves ethanolic extract has antigenotoxic effect shown by decreasing of the buccal micronucleus frequency of rat.

ABSTRAK

Polycyclic aromatic hydrocarbon atau PAH merupakan salah satu kelompok karsinogen terbesar di lingkungan. 7,12-Dimetillbez(α)antransena merupakan senyawa golongan PAH yang bersifat karsinogen genotoksik. Salah satu uji genotoksisitas yang paling sering dilakukan adalah uji mikronukleus. Sirsak merupakan tanaman yang tumbuh baik di Indonesia. Daun sirsak mengandung avonoid dan acetogenin yang diduga mempunyai potensi kemopreventif dan aktivitas antikanker. Tujuan penelitian ini adalah mengkaji efek antigenotoksik ekstrak etanolik daun sirsak terhadap frekuensi mikronukleus mukosa bukal tikus galur Sprague Dawley yang diinduksi dengan DMBA. Penelitian ini dilakukan pada 24 tikus Sprague Dawley jantan berumur 5 minggu yang dibagi secara acak dalam 6 kelompok. Karsinogenesis dorsum lidah tikus kelompok I – III diinduksi dengan DMBA secara topikal 3 kali dalam seminggu selama 16 minggu, kelompok II dan III selain diinduksi karsinogenesis, juga diberi ekstrak etanolik daun sirsak 100 dan 200 mg/kg BB setiap hari selama 18 minggu dan kelompok IV diberi ekstrak etanolik daun sirsak 200 mg/kg BB, kelompok V diberi DMSO 1% dan kelompok VI tidak diberi perlakuan. Setelah minggu ke-18, swab mukosa bukal dilakukan untuk uji mikronukleus kemudian sampel dicat dengan metode Feulgen-Rossenbeck. Jumlah mikronukleus dihitung di bawah mikroskop cahaya per 500 sel epitel mukosa bukal, lalu data dianalisis menggunakan one way ANOVA diikuti Tukey HSD. Hasil penelitian menunjukkan bahwa rerata frekuensi mikronukleus kelompok II (13 ± 0,82) dan kelompok III (12 ± 0,96) mengalami penurunan secara signikan (p<0,05) dibanding kelompok I (24 ± 1,71). Disimpulkan bahwa ekstrak etanolik daun sirsak mempunyai efek antigenotoksik yang ditunjukkan dengan penurunan frekuensi mikronukleus sel epitel mukosa bukal tikus.

Keywords

DMBA; soursop leaves ethanolic extract; micronucleus frequency; PAH

Full Text:

PDF

References

US. Department of health and human services. Report on Carcinogens, 12th Ed. Public Health Service, National Toxicology Program. 2011.

Sen DJ, Shisio CJ, Lahiri A. Three musketeers of genotoxicity: carcinogen, mutagen and teratogen. NSHM Journal of Pharmacy and Healthcare Management. 2011; 2: 13 – 25.

King RJB. Cancer biology, 2nd Ed. London: Pearson Education; 2000.Tudoran MA, Putz MV. Polycyclic aromatic hydrocarbons: from in cerebro to in silico eco- toxicity fate, chem. Bull. “Politehnica” Univ, (Timisoara). 2012; 57(71): 50 – 53.

IPCS. Environmental health criteria no. 202. selected non-heterocyclic polycyclic aromatic hydrocarbons. international programme on chemical safety. 1998. http://www.inchem.org/documents/ehc/ehc/ehc202.htm, diunduh 9 September 2014.

Okamura M, UnamiA, Moto M, Maqumuru M, Ito T, Jin M, Kashida Y, Mitsumori K. The possible mechanism of enhanced carcinogenesis induced by genotoxic carcinogens in rasH2 mice. Cancer Lett. 2007; 245(1-2): 321 – 30.

Kitakawa D, Cabral LAG, Marques MEA, Salvadori DMF, Ribeiro DA. Medium- term tongue carcinogenesis assays: A comparative study between 4-nitroquinoline 1-oxide (4NQO)-induced rat and dimethylbenzanthracene (DMBA)-induced hamster carcinogenesis. JEANS. 2006; 43: 219 – 227.

Sun Z, Sood S, Li N, Yang P, Newman RA, Yang CS, Chen X. Chemoprevention of 7, 12-dimethylbenz[a]anthracene (DMBA) - induced oral carcinogenesis in hamster cheek pouch by topical application of a dual inhibitor of epidermal growth factor receptor (EGFR) and ErbB2 tyrosine kinases. Oral Oncol. 2008; 44(7): 652 – 657.

Yu T, Wang X, Gong R, Li A, Yang S, Cao Y, Wen Y, Wang C, Yi X. The expression prole of microRNAs in a model of 7,12-dimethyl- benz[a]anthrance-induced oral carcinogenesis in Syrian hamster. JECCR. 2009; 28(64): 1 – 10.

Berta GN, Salamone P, Sprio AE, Scipio FD, Marinos LM, Sapin S, Carlotti ME,
Cavalli R, Carlo FD. Chemoprevention of 7,12-dimethylbenz[a]anthracene (DMBA)- induced oral carcinogenesis in hamster cheek pouch by topical application of resveratrol complexed with 2-hydroxypropyl- b-cyclodextrin. Oral Oncol. 2010; 46: 42 – 48.

Hsu WH, Lee BH, Pan TM. Effects of red mold dioscorea on oral carcinogenesis in dmba- induced hamster animal model. Food Chem Toxicol. 2011; 49: 1292 – 1297.

Balakrisnan S, Menon VP, Manoharan S, Rajalingan K. Antigenotoxic effect of ferulic acid in 7, 12-dimethyl benz(α)anthancene induced genotocicity. Afr. J. Traditional. 2008; 5(1): 32 – 38.

Bolt HM, Stewart JD, Hengstler JG. A comprehensive review about micronuclei: mechanisms of formation and practical aspects in genotoxicity testing. Arch Toxicol. 2011; 85: 861 – 862.

Melendez-Colon VJ, Luch A, Seidel A, Baird WM. Cancer initiation by polycyclic aromatic hydrocarbon results from formation of stable DNA Adducts rather than apurinic sites. Carcinogenesi. 1999; 20(10): 1885 – 1891.

Ray G, Husain SA. Oxidant, antioxidant and carcinogenesis. IJEB. 2002; 40: 1213 – 1232.

Igarashi M, Nagata M, Itoh S, Yamoto T, Tsuda S. Relationship between DNA damage and micronucleus in mouse liver. J. Toxicol. Sci. 2010; 35(6): 881 – 889.

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; 26(1): 125 – 132.

Holland N, Bolognesi C, Bonassi S, Zeiger E, Fenech M, Knasmueller S. The micronucleus assay in human buccal cells as a tool for biomonitoring DNA damage: the HUMN project perspective on current status and knowledge gaps. Mut. Res. 2008; 659(1-2): 93 – 108.

Tanaka T, Ishigamori R. Understanding carcinogenesis for ghting oral cancer: Riview Article. Journal of Oncology. 2011. 1 – 10.

Żelazna K, Rudnicka K, Tejs S. In vitro micronucleus test assessment of polycyclic aromatic hydrocarbons. Environmental Biotechnology. 2011; 7(2): 70 – 80.

Harsvardhan SJ, Alka DK, Mohan KKP. Micronucleus as potential biomarker of oral carcinogenesis. IJDA. 2010; 2(2): 197 – 202.

Hamizah S, Roslida AH, Fezah O, Tan KL, Tor YS, Tan CI. Chemopreventive potential of Annona muricata L. leaves on chemically- induced skin papillomagenesis in mice. APJP. 2012; 13: 2533 – 2539.

Baskar R, Rajeswari V, Kumar TS. In vitro antioxidant studies in leaves of annona species. Indian J. Exp Biol. 2007; 45(5): 480 – 485.

Moghadamtousi ZS, Karimian H, Rouhollahi E, Paydar M, Fadaeinasab M, Kadir HA. Annona muricata leaves induce G₁ cell cycle arrest and apoptosis through mitochondria-mediated pathway in human HCT-116 and HT-29 colon cancer cells. J Ethnopharmacol. 2014; 156: 277 – 289.

Torres MP, Rachagani S, Purohit V, Pandey P, Joshi S, Moore ED, Johansson SJ, Singh PK, Ganti AK, Batra SK. Graviola: A novel promising natural-derived drug that inhibits tumorigenicity and metastasis of pancreatic cancer cells in vitro and in vivo through altering cell metabolism. Cancer Lett. 2012; 323(1): 29 – 40.

Minari JB, Okeke U. Chemopreventive effect of Annona muricata on DMBA-induced cell proliferation in the breast tissues of female albino mice. The Egypt J of Med. Hum Genet. 2014; 15(4): 327 – 334.

Asare GA,Afriye D, Ngala RA, Abutiate H, Doku D, Mahmood SA, Rahman H. Antiproliferative activity of aqueous leaf extract of Annona muricata L. on the prostate, BPH-1 cells, and some target genes. Integr Cancer Ther. 2015; 14(1): 65 – 74.

Moghadamtousi ZS, Fadaeinasab M, Nikzad S, Mohan G, Ali HM, Kadir HA. Annona muricata (Annonaceae): A review of its traditional uses, isolated acetogenins and biological activities. Int. J Mol Sci. 2015; 16: 15625 – 15658.

Shantiningsih RR. Patch gingiva mucoadesive β-carotene sebagai pencegah efek samping paparan radiasi radiogra panoramik (kajian in vivo pada kelinci galur New Zealand. Disertasi. Yogyakarta Universitas: Gadjah Mada. 2014.

Dindgire SL, Gosavi S, Kurmawat RM, Ganvir S, Hazarey V. Comparative study of exfoliated oral mucosal cell micronucleus frequency in potentially malignant and malignant lesions. IJOMP. 2012; 3(2): 15 – 20.

Buajeeb W, Kraivaphan P, Amornchat C, Suthamajaya K. Reduction of micronuclei in oral lichen planus supplemented with beta- carotene. JOS. 2008; 50(4): 461 – 467.

Tolbert PE, Shy CM, Allen JW. Micronuclei and other nuclear anomalies in buccal smears: methods development. Mutant Res. 1992; 271(1): 69 – 77.

Panjamurthy K, Manoharan S, Balakrisnan S, Suresh K, Nirmal MR, Senthi N, Alias M. Protectiveeffectofwithaferin-aonmicronucleus frequency and detoxication agents during experimental oral carcinogenesis. J. Trad. CAM. 2009; 6(1): 1 – 8.

Choudhari SK, Chaudhary M, Bagde S, Gadbail AR, Josh V. Nitric Oxide and Cancer: A Review. World Jurn. of Surg. Oncol. 2013; 11: 118.

Pieme CA, Kumar SG, Dongmo MS, Moukette BM, Boyoum FF, Ngogang JY, Saxena AK. Antiproliferative activity and induction of apoptosis by Annona muricata (Annonaceae) extract on human cancer cells. BMC. 2014. 516.

Batra P, Sharma AK. Anti-cancer potential of avonoids: recent trends and future perspectives. Biotech. 2013; 3: 439 – 459.

Chahar MK, Sharma N, Dobhal MP, Joshi YC. Flavonoids: a versatile source of anticancer drugs. Pharmacogn Rev. 2011; 5(9): 1 – 12.

Shah S.Kaur M. Biomarkers and chemopreventives in oral carcinogenesis and its prevention. Journal of Oral and Maxillofacial Pathology. 2014; 18(1): 114 – 121.

Arthur FKN, Woode E, Larbie C, Terlabi EO. Evaluation of acute and subchronic toxicity of Annona muricata (Linn) aquaeus extract in animals. Euro. J.Exp. Bio. 2011; 1(4): 115 – 124.

DOI: https://doi.org/10.22146/majkedgiind.11794