نوع مقاله : مقاله پژوهشی
نویسندگان
1 گروه زیست شناسی دریا، دانشکده علوم دریایی و اقیانوسی، دانشگاه علوم و فنون دریایی خرمشهر، خرمشهر، ایران.
2 گروه زیست شناسی دریا، دانشکده علوم دریایی و محیطی، دانشگاه مازندران، بابلسر، ایران.
3 گروه آناتومی و جنین شناسی، مرکز تحقیقات سلولی و مولکولی خلیج فارس، دانشگاه جندی شاپور اهواز، ایران
چکیده
کبد یکی از مهم ترین اندام های بدن جانوران محسوب می شود که به عنوان اندام سم زدا و پالایشگر سموم بدن شناخته می شود و در اغلب مطالعات توکسیکولوژیک به عنوان اندام هدف مورد استفاده قرار می گیرد. برای انجام این مطالعه، از 8 عدد ماهی هامور معمولی نابالغ (نر و ماده به تعداد مساوی) استفاده شد. سپس ماهی بیهوش و سر آن ها از بدن جدا شد. بدن ماهی توسط الکل اتانول 70% ضدعفونی و پس از جدا کردن بافت کبد، به قطعات ریز تقسیم و توسط آنزیم کلاژناز تیپ 4 هضم شدند. سلول های کبدی به مدت 2 هفته در دمای c °30 و میزان 20% (FBS: Fetal bovine serum) به همراه ITS (Insulin- transferin- selenium) کشت داده شدند، سپس دوزهای آلودگی (µM 100، µM 100*2 ،µM100*3) به نمونه ها اضافه و در زمان های صفر، 6، 12 و 24 ساعت مطالعه شدند. مرفولوژی سلول ها توسط میکروسکوپ معکوس و درصد بقا توسط سنجش MTT (Methyl thiazolyl tetrazolium: متیل تیازول تترازولیوم) ارزیابی گردید. آنالیز مرفولوژی نشان داد که دوز 100µm موجب القا 50 درصد مرگ سلولی می شود و به کارگیری غلظت های پایین آلاینده بنزوآلفاپایرن طی تیمار 24 ساعت اثر مهار تکثیر وابسته به دوز و زمان داشته و در دوزهای بالاتر باعث لیز شدن سلولها و نکروزیس می شود.
کلیدواژه‌ها
موضوعات
Au, D.W.T., 2004. The application of histo-cytopathological biomarkers in marine pollution monitoring: a review. Marine pollution bulletin, 48(9-10), pp.817-834.
Babich, H. and Borenfreund, E., 1991. Cytotoxicity and genotoxicity assays with cultured fish cells: a review. Toxicology in vitro, 5(1), pp.91-100.
Banaee, M., Sureda, A., Mirvaghefi, A.R. and Ahmadi, K., 2011. Effects of diazinon on biochemical parameters of blood in rainbow trout (Oncorhynchus mykiss). Pesticide biochemistry and physiology, 99(1), pp.1-6. (In Persian)
Benson, W. and Schlenk, D., 2001. Target Organ Toxicity in Marine and Freshwater Teleosts, Volume 1-Organs. Taylor & Francis.
Borenfreund, E. and Puerner, J.A., 1985. Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicology letters, 24(2-3), pp.119-124.
Brüschweiler, B.J., Würgler, F.E. and Fent, K., 1995. Cytotoxicity in vitro of organotin compounds to fish hepatoma cells PLHC-1 (Poeciliopsis lucida). Aquatic toxicology, 32(2-3), pp.143-160.
Caminada, D., Escher, C. and Fent, K., 2006. Cytotoxicity of pharmaceuticals found in aquatic systems: comparison of PLHC-1 and RTG-2 fish cell lines. Aquatic toxicology, 79(2), pp.114-123.
Castaño, A. and Becerril, C., 2004. In vitro assessment of DNA damage after short-and long-term exposure to benzo (a) pyrene using RAPD and the RTG-2 fish cell line. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 552(1-2), pp.141-151.
Castaño, A. and Gómez-Lechón, M.J., 2005. Comparison of basal cytotoxicity data between mammalian and fish cell lines: a literature survey. Toxicology in Vitro, 19(5), pp.695-705.
Chae, Y.J., Pham, C.H., Lee, J., Bae, E., Yi, J. and Gu, M.B., 2009. Evaluation of the toxic impact of silver nanoparticles on Japanese medaka (Oryzias latipes). Aquatic toxicology, 94(4), pp.320-327.
Chi, S.C., Hu, W.W. and Lo, B.J., 1999. Establishment and characterization of a continuous cell line (GF‐1) derived from grouper, Epinephelus coioides (Hamilton): a cell line susceptible to grouper nervous necrosis virus (GNNV). Journal of Fish Diseases, 22(3), pp.173-182.
Dabiri, M., 2013. Environmental pollution (air-water-soil-sound), Etihad Publication. Tehran. 399 p.
Di Giulio, R.T. and Hinton, D.E., 2008. The toxicology of fishes. Crc Press.
Esmaili Sari, A., 2002. Pollutants, health and standards in the environment. Naqsh Mehr Publishing, Tehran. 767 p.
Fent, K. and Hunn, J., 1996. Cytotoxicity of organic environmental chemicals to fish liver cells (PLHC-1). Marine Environmental Research, 42(1-4), pp.377-382.
Fent, K., 2001. Fish cell lines as versatile tools in ecotoxicology: assessment of cytotoxicity, cytochrome P4501A induction potential and estrogenic activity of chemicals and environmental samples. Toxicology in vitro, 15(4-5), pp.477-488.
Fernandes, C., Fontaínhas-Fernandes, A., Rocha, E. and Salgado, M.A., 2008. Monitoring pollution in Esmoriz–Paramos lagoon, Portugal: Liver histological and biochemical effects in Liza saliens. Environmental Monitoring and Assessment, 145, pp.315-322.
Forth, W., Henschler, D., Rummel, W. and Starke, K., 2001. Allgemeine und spezielle Pharmakologie und Toxikologie. Aufl. München: Urban und Fischer.
Genten, F., Terwinghe, E. and Danguy, A., 2009. Atlas of fish histology. Science publisher, 92-98, 215p.
Gernhöfer, M., Pawert, M., Schramm, M., Müller, E. and Triebskorn, R., 2001. Ultrastructural biomarkers as tools to characterize the health status of fish in contaminated streams. Journal of Aquatic Ecosystem Stress and Recovery, 8, pp.241-260.
Health, A. G., 1995. Water Pollution and Fish Physiology, seconded. CRC Lewis Publishers, Boca Ratonal, pp.125-140.
Hellawell, J. M., 1986. Biological indicators of freshwater pollution and environmental stressors on fisheshes. Elsevier Applied Science Publishers, 8, pp.51-66.
Hinton, D. E. and Lauren, D. J., 1990. Integrative histopathological approaches to detecting effects of environmental stressors on fishes. In: Adams, S. M. (Ed.), Biological Indicators of Stress in Fish: American Fisheries Symposium 8. American Fisheries Society, Bethesda, Maryland, 51-66.
Kemnitzer, W., Drewe, J., Jiang, S., Zhang, H., Zhao, J. and Crogan- Grundy, C., 2007. Discovery of 4-Aryl-4 H-chromenes as a New Series of Apoptosis Inducers Using a Cell- and Caspase- Based High- Throughput Screening Assay. 3. Structure- Activity Relationships of Fused Rings at the 7. 8- Positions. Jurnal of Medical Chemistry, 50 (12), pp.2858-64.
Landis, W.G., Sofield, R.M. and Yu, M.H., 2017. Introduction to environmental toxicology: molecular substructures to ecological landscapes. CRC Press.
Laville, N., Aıt-Aıssa, S., Gomez, E., Casellas, C. and Porcher, J.M., 2004. Effects of human pharmaceuticals on cytotoxicity, EROD activity and ROS production in fish hepatocytes. Toxicology, 196(1-2), pp.41-55.
Law, J.M., 2003. Issues related to the use of fish models in toxicologic pathology: session introduction. Toxicologic pathology, 31(1_suppl), pp.49-52.
Li, H. and Zhang, S., 2002. In vitro cytotoxicity of the organophosphorus insecticide methylparathion to FG-9307, the gill cell line of flounder (Paralichthys olivaceus). Cell Biology and Toxicology, 18, pp.235-241.
Momeni, H. R., Soliemani Mehranjani, M., Abnosi, M. H. and Nasimi, P., 2010. Evaluation of viability in adult mouse spinal cord slices by MTT. Iranian Journal of biology, 23(2), pp. 267-274. (In Persian)
Mosmann, T., 1983. Rapid colorimetric assay for cellular growth and survival application to proliferation and cytotoxicity assays. Journal of Immunolgiae Methods, 63. pp.55-65.
Ogundiran, M.A., Fawole, O.O., Adewoye, S.O. and Ayandiran, T.A., 2010. Toxicological impact of detergent effluent on juvenile of African Catfish (Clarias gariepinus)(Buchell, 1822). Agriculture and biology journal of North America, 1(3), pp.330-342.
Oliveira, M., Pacheco, M. and Santos, M.A., 2008. Organ specific antioxidant responses in golden grey mullet (Liza aurata) following a short-term exposure to phenanthrene. Science of the Total Environment, 396(1), pp.70-78.
Parameswaran, V., Ishaq Ahmed, V.P., Shukla, R., Bhonde, R.R. and Sahul Hameed, A.S., 2007. Development and characterization of two new cell lines from milkfish (Chanos chanos) and grouper (Epinephelus coioides) for virus isolation. Marine Biotechnology, 9, pp.281-291.
Reynaud, S. and Deschaux, P., 2006. The effects of polycyclic aromatic hydrocarbons on the immune system of fish: a review. Aquatic toxicology, 77(2), pp.229-238.
Roesijadi, G. and Robinson, W. E., 1994. Metal regulation in aquatic animals: mechanism of uptake, accumulation and release. In: Baykan, U., Atli, G. and Canli, M., 2007. The effects of temperature and metal exposures on the profiles of metallothionein-like proteins in Oreochromis niloticus. Environmental Toxicology and Pharamacology, 23, pp.33-38.
Romijn, J.C., Verkoelen, C.F. and Schroeder, F.H., 1988. Application of the MTT assay to human prostate cancer cell lines in vitro: establishment of test conditions and assessment of hormone‐stimulated growth and drug‐induced cytostatic and cytotoxic effects. The prostate, 12(1), pp.99-110.
Roy, S., Chattoraj, A. and Bhattacharya, S., 2006. Arsenic-induced changes in optic tectal histoarchitecture and acetylcholinesterase–acetylcholine profile in Channa punctatus: amelioration by selenium. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 144(1), pp.16-24.
Rohn, M., Tenhagen, B.A. and Hofmann, W., 2004. Survival of dairy cows after surgery to correct abomasal displacement: 1. Clinical and laboratory parameters and overall survival. Journal of Veterinary Medicine Series A, 51(6), pp.294-299.
Schirmer, K., Dixon, D.G., Greenberg, B.M. and Bols, N.C., 1998. Ability of 16 priority PAHs to be directly cytotoxic to a cell line from the rainbow trout gill. Toxicology, 127(1-3), pp.129-141.
Sikkema, J., de Bont, J.A. and Poolman, B., 1994. Interactions of cyclic hydrocarbons with biological membranes. Journal of biological Chemistry, 269(11), pp.8022-8028.
Stentiford, G.D., Longshaw, M., Lyons, B.P., Jones, G., Green, M. and Feist, S.W., 2003. Histopathological biomarkers in estuarine fish species for the assessment of biological effects of contaminants. Marine Environmental Research, 55(2), pp.137-159.
Suresh, N., 2009. Effect of cadmium chloride on liver, spleen and kidney melano macrophage centres in Tilapia mossambica. Journal of environmental biology, 30(4), pp.505-508.
Taju, G., Majeed, S.A., Nambi, K.S.N., Farook, M.A., Vimal, S. and Hameed, A.S., 2014. In vitro cytotoxic, genotoxic and oxidative stress of cypermethrin on five fish cell lines. Pesticide Biochemistry and Physiology, 113, pp.15-24.
Taju, G., Majeed, S.A., Nambi, K.S.N. and Hameed, A.S., 2014. In vitro assay for the toxicity of silver nanoparticles using heart and gill cell lines of Catla catla and gill cell line of Labeo rohita. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 161, pp.41-52.
Thophon, S., Kruatrachue, M., Upatham, E.S., Pokethitiyook, P., Sahaphong, S. and Jaritkhuan, S., 2003. Histopathological alterations of white seabass, Lates calcarifer, in acute and subchronic cadmium exposure. Environmental pollution, 121(3), pp.307-320.
Van Dyk, J.C., Pieterse, G.M. and Van Vuren, J.H.J., 2007. Histological changes in the liver of Oreochromis mossambicus (Cichlidae) after exposure to cadmium and zinc. Ecotoxicology and environmental safety, 66(3), pp.432-440.
Weber, L.P. and Janz, D.M., 2001. Effect of β-naphthoflavone and dimethylbenz [a] anthracene on apoptosis and HSP70 expression in juvenile channel catfish (Ictalurus punctatus) ovary. Aquatic Toxicology, 54(1-2), pp.39-50.
Velma, V. and Tchounwou, P.B., 2010. Chromium-induced biochemical, genotoxic and histopathologic effects in liver and kidney of goldfish, Carassius auratus. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 698(1-2), pp.43-51.
Wen, C.M., Lee, C.W., Wang, C.S., Cheng, Y.H. and Huang, H.Y., 2008. Development of two cell lines from Epinephelus coioides brain tissue for characterization of betanodavirus and megalocytivirus infectivity and propagation. Aquaculture, 278(1-4), pp.14-21.
Zhou, B., Liu, C., Wang, J., Lam, P.K. and Wu, R.S., 2006. Primary cultured cells as sensitive in vitro model for assessment of toxicants-comparison to hepatocytes and gill epithelia. Aquatic toxicology, 80(2), pp.109-118.
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