بررسی آزمایشگاهی اثر سایتوتوکسیسیتی آفلاتوکسین‌هایB1 و G1 برروی لنفوسیت‌های T موشی

نوع مقاله : مقاله کامل

نویسندگان

1 گروه بیوشیمی، دانشگاه پیام نور، اصفهان، ایران

2 گروه زیست شناسی، دانشگاه پیام نور، تهران، ایران

3 موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران

چکیده

آفلاتوکسین‌ها ترکیبات شیمیائی سمی مشتق از قارچ هستند که توسط سویه‌هایی از قارچ آسپرژیلوس تولید می‌شوند. این مطالعه با هدف جداسازی آفلاتوکسین‌های ناشی از آسپرژیلوس‌های مولد آفلاتوکسین جداشده ازخوراک دام و بررسی سمیت سلولی (سایتوتوکسیسیتی) آن‌ها برروی رده سلولی لنفوسیت T موش انجام شده است. در این تحقیق از قارچ‌های آسپرژیلوس فلاووس (Aspergillus flavus) و آسپرژیلوس پارازیتیکوس(Aspergillus parasiticus) استاندارد و جدا شده از خوراک دام استفاده شد. قابلیت تولید آفلاتوکسین توسط این سویه‌ها به روش‌های کیفی (TLC) و کمی (HPLC) مورد بررسی قرارگرفت. سپس رده سلولی لنفوسیت T موش (EL4) درمواجهه با غلظت‌های مختلف آفلاتوکسین قرار گرفت و میزان توکسیسیتی آن به روش MTT اندازه‌گیری شد. میزان سلول‌های کشته شده نیز به روش فلوسایتومتری ارزیابی شد. بیشترین میزان تولید آفلاتوکسین G1 و B1 در سویه استاندارد آ. پارازیتیکوس و آ. فلاووس جدا شده از خوراک دام و بیشترین میزان توکسی سیتی آفلاتوکسین‌های فوق در رقت 1/0 از آفلاتوکسین مشتق از آ. پارازیتیکوس مشاهده شد. نتایج به دست آمده در این تحقیق نشان داد که سویه‌های آسپرژیلوس جدا شده از خوراک دام قابلیت تولید آفلاتوکسین‌های G1 و B1 رادارند که اثر کشندگی قابل ملاحظه‌ای بر روی رده لنفوسیتی T موش نشان دادند که این امر می‌تواند بیانگر اهمیت حفظ سلامت و بهداشت غذای دام باشد.

کلیدواژه‌ها


عنوان مقاله [English]

The In-vitro Cytotoxicity Effect of Aflatoxins B1 and G1 on Mice T lymphocyte

نویسندگان [English]

  • N. Keikha 1
  • N. Sohrabi 2
  • M. Taghizadeh 3
1 Department of Biochemistry, Payame Noor University, Tehran, Iran
2 Department of Biology, Payame Noor University, Tehran, Iran
3 Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extention Organization (AREEO), Karaj, Iran
چکیده [English]

Aflatoxins are chemical toxic metabolites which are produced by Aspergillus species. This study was designed to investigate the aflatoxines cytotoxicity effects on mice T lymphocyte. In this work, four different standard and feed derived strains of A. parasiticus and A. flavus were selected. The levels of G1 and B1 aflatoxins were determined by TLC and HPLC methods. For evaluation of cytotoxity effect, the different concentration of aflatoxins were mixed by mice T cell line (EL4) on different time and the percentage of cell death were been evaluated by MTT assay and flowcytometry (FCM) method. Among different strains, the standard A. parasiticus and feed derived A. flavus have highest levels of G1 and B1. The most cytotoxity effect were seen on 1/10 concentration of A. parasiticus extracted aflatoxins. These results shown feed derived Aspergillus could produce G1 and B1 aflatoxins which have considerable cytotoxicity effect on mice T lymphocytes.

کلیدواژه‌ها [English]

  • Aspergillus
  • Aflatoxin
  • Cytotoxicity
  • Cattle feed
  • T lymphocyte
1- Abbes, S., Ben Salah-Abbes, J., Abdel-Wahhab, M.A., Ouslati, R. 2010. Immunotoxicological and biochemical effects of aflatoxins in rats prevented by Tunisian montmorillonite with reference to HSCAS. Immunopharmacology and Immunotoxicology, 32(3):514-522.
2- Bruneau, J.C., Stack, E., O'Kennedy, R., Loscher, C.E. 2012. Aflatoxins B (1), B (2) and G (1) modulate cytokine secretion and cell surface marker expression in J774A.1 murine macrophages. Toxicology in Vitro, 26(5):686-93.
3- Eaton, D.L., Gallagher, E.P. 1994. Mechanisms of aflatoxin carcinogenesis. Annual Review of Pharmacology and Toxicology, 34:135-72.
4- Gnonlonfin, G.J., Hell, K., Adjovi, Y., Fandohan, P., Koudande, D.O., Mensah, G.A. 2013. A review on aflatoxin contamination and its implications in the developing world: a sub-Saharan African perspective. Critical Reviews in Food Science and Nutrition, 53(4):349–65.
5- Hinton, D.M., Myers, M.J., Raybourne, R.A., Francke-Carroll, S., Sotomayor, R.E., Shaddock, J., Warbritton, A., Chou, M.W. 2003. Immunotoxicity of aflatoxin B1 in rats: effects on lymphocytes and the inflammatory response in a chronic intermittent dosing study. Toxicological Sciences, 73(2):362-77.
6- Hou, Y.J., Zhao, Y.Y., Xiong, B., Cui, X.S., Kim, N.H., Xu, Y.X., Sun, S.C. 2013. Mycotoxin-containing diet causes oxidative stress in the mouse. PLoS One, 8(3):e60374.
7- Jiang, M., Peng, X., Fang, J., Cui, H., Yu, Z., Chen, Z. 2015. Effects of aflatoxin b1 on T-cell subsets and mRNA expression of cytokines in the intestine of broilers. International Journal of Molecular Sciences, 16(4):6945-59.
8- Kumar, P., Mahato, D.K., Kamle, M., Mohanta, T.K., Kang, S.G. 2017. Aflatoxins: A Global Concern for Food Safety, Human Health and Their Management. Frontiers in microbiology, 7:2170.
9- Lewis, C., Smith, J., Anderson, J., Freshney, R. 1999. Increased cytotoxicity of food borne mycotoxins toward human cell lines in vitro via enhanced cytochrome P450 expression using the MTT bioassay. Mycopathologia, 148(2):97-102.
10- Luongo, D., Russo, R., Balestrieri, A., Marzocco, S., Bergamo, P., Severino, L. 2013. In vitro study of AFB1 and AFM1 effects on human lymphoblastoid Jurkat T-cell model. Journal of Immunotoxicology, 11:353–358.
11- Marchese, S., Polo, A., Ariano, A., Velotto, S., Costantini, S., Severino, L. 2018. Aflatoxin B1 and M1: Biological Properties and Their Involvement in Cancer Development. Toxins (Basel), 10(6).pii: E214
12- Meissonnier, G.M., Pinton, P., Laffitte, J., Cossalter, A.M., Gong, Y.Y., Wild, C.P., Bertin, G., Galtier, P., Oswald, I.P. 2008. Immunotoxicity of aflatoxin B1: impairment of the cell-mediated response to vaccine antigen and modulation of cytokine expression. Toxicology and Applied Pharmacology, 231(2):142-9.
13- Mohsenzadeh, M.S., Hedayati, N., Riahi-Zanjani, B., Karimi, G. 2016. Immunosuppression following dietary aflatoxin B1 exposure: a review of the existing evidence. Toxin Reviews, 35:121–127.
14- Moon, E.Y., Rhee, D.K., Pyo, S. 1999. In vitro suppressive effect of aflatoxin B1 on murine peritoneal macrophage functions. Toxicology, 133(2-3):171-9.
15- Rahimi, S., Sohrabi, N., Ebrahimi, M.A., Tebyanian, M., Taghizadeh, M. 2016. Studying the effect of aflatoxin genes Aflp and Aflq on Aspergillus flavus and Aspergillus parasiticus in the cattle feed used in industrial animal husbandries. Acta Medica Mediterranea, 32(6):2091-2100.
16- Raisuddin, S., Singh, K.P., Zaidi, S.I., Paul, B.N., Ray, P.K. 1993. Immunosuppressive effects of aflatoxin in growing rats. Mycopathologia, 124(3):189-94.
17- Rushing, B.R., Selim, M.I. 2019. Aflatoxin B1: A review on metabolism, toxicity, occurrence in food, occupational exposure, and detoxification methods. Food and Chemical Toxicology, 124:81-100.
18- Shen, H., Shi, C., Shen, Y.I., Ong, C. 1996. Detection of elevated reactive oxygen species level in cultured rat hepatocytes treated with aflatoxin B1. Free Radical Biology & Medicine, 21(2):139-146.
19- Shirani, K., Zanjani, B.R., Mahmoudi, M., Jafarian, A.H., Hassani, F.V., Giesy, J.P, Karimi G. 2018. Immunotoxicity of aflatoxin M1 : as a potent suppressor of innate and acquired immune systems in a subacute study. Journal of the Science of Food and Agriculture, 98(15):5884-5892.
20- Sohrabi, N., Taghizadeh, M. 2018. Molecular identification of aflatoxigenic Aspergillus species in feedstuff samples. Current Medical Mycology, 4(2):1-6.
21- Tomková, I., Sevcíková, Z., Levkut, M., Revajová, V., Conková, E., Laciaková, A., Lenhardt, L. 2002. Effect of aflatoxin B1 on CD3 T cells and alkaline phosphatase in the intestine of mice. Mycopathologia, 154(1):15-9.
22- Umesha, S., Manukumar, H.M., Chandrasekhar, B., Shivakumara, P., ShivaKumar, J., Raghava, S., Avinash, P., Shirin, M., Bharathi, T.R., Rajini, S.B., Nandhini, M., Vinaya Rani, G.G., Shobha, M., Prakash, H.S. 2017. Aflatoxins and food pathogens: impact of biologically active aflatoxins and their control strategies. Journal of the Science of Food and Agriculture, 97(6):1698-1707.
23- Wild, C.P., Turner, P.C. 2002. The toxicology of aflatoxins as a basis for public health decisions. Mutagenesis, 17(6):471-81.
24- Williams, J., Phillips, T., Jolly, P., Stiles, J., Jolly, C., Aggarwal, D. 2004. Human aflatoxicosis in developing countries: a review of toxicology, exposure, Potential health consequences, and interventions. The American Journal of Clinical Nutrition, 80(5):1106–22.
25- Zimmermann, C.E.P., Machado, A.K., Cadoná, F.C., Jaques, J.A.S., Schlemmer, K.B., Lautert, C., Cruz, I.B.M., Zanette, R.A., Leal, D.B.R., Santurio, J.M. 2014. Invitro cytotoxicity of aflatoxin B1 to broiler lymphocytes of broiler chickens. Brazilian Journal of Poultry Science, 16(3), 307-312.