اثرات نانوذرات اکسید روی بر بیان ژن‌‌های انتقال‌دهنده روی 1-4 در سل لاین هیپوکامپ موش‌‌های صحرایی نر
محورهای موضوعی : سلولی ملکولی
مائده نیله چی
1
(دانشجوی دکتری، گروه زیست‌شناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران.)
اکرم عیدی
2
(استاد، گروه زیست‌شناسی، واحد علوم و تحقیقات، دانشگاه آزاد اسلامی، تهران، ایران)
حمید گله داری
3
(استاد، گروه زیست‌شناسی، دانشگاه شهید چمران اهواز، اهواز، ایران)
مهناز کسمتی
4
(استاد، گروه زیست‌شناسی، دانشگاه شهید چمران اهواز، اهواز، ایران.)
کلید واژه: هیپوکامپ, Znt, ZnO, موش‌‌های صحرایی, Real-Time RT-PCR, نانوذرات, بیان ژن,
چکیده مقاله :
سابقه و هدف: عنصر روی نقش مهمی در کارکرد ارگان‌‌های حیاتی، به ویژه سیستم عصبی مرکزی ایفا می‌‌نماید. اختلال در هموستازی روی موجب ایجاد و پیشرفت بیماری‌‌های سیستم عصبی نظیر آلزایمر، افسردگی، اختلال در یادگیری و استرس می‌‌گردد. هموستازی روی در بدن توسط پروتئین‌‌های ZnT و ZIP صورت می‌‌پذیرد. هدف مطالعه حاضر بررسی اثر نانوذرات اکسید روی در بیان ژن‌‌های انتقال‌دهنده روی 1-4 موسوم به Znt1،Znt2، Znt3 و Znt4در سلول‌‌های هیپوکامپ به عنوان یکی از بافت‌‌هایی که تراکم بالایی از روی را در خود جای داده، می‌‌باشد. روش کار: ابتدا پاساژ سلولی رده سلولی هیپوکامپ انجام شد، سپس تست MTT assay برای نانوذره اکسید روی صورت گرفت. در مرحله بعد استخراج RNA و سنتز CDNA انجام شده و جهت اطمینان از خلوص نمونه RNA از اسپکتروفتومتر نانودراپ استفاده گردید و پرایمرهای اختصاصی و مناسب ژن‌‌های مورد نظر طراحی و سنتز شد. سپس با استفاده از Real-Time PCR تغییرات بیان ژن‌‌های Znt1، Znt2، Znt3 و Znt4 بررسی گردید. یافته‌‌ها: غلظت‌‌های 10 و 20 μg/mL از نانوذره اکسید روی ضمن ایجاد کم‌ترین سیتوتوکسیتی، موجب افزایش بیان معنادار هر چهار ژن Znt1،Znt2، Znt3 و Znt4 در رده سلولی هیپوکامپ موش صحرایی گردید. نتیجه‌گیری: نانوذره اکسید روی می‌‌تواند با افزایش بیان ژن‌‌های انتقال‌دهنده روی Znt1،Znt2، Znt3 و Znt4 در درمان بیماری‌‌های ناشی از اختلال هموستازی روی نظیر آلزایمر، افسردگی، اختلال در یادگیری و استرس مورد بررسی داروشناسی قرار گیرد.
Introduction: Zinc plays an important role in the function of vital organs, especially the central nervous system. Zinc homeostasis disorder causes and progresses nervous system diseases such as Alzheimer, depression, learning disabilities and stress. Zinc homeostasis in the body is mediated by ZnT and ZIP proteins. The aim of this study was to investigate the effect of zinc oxide nanoparticles on the expression of Znt1, Znt2, Znt3, and Znt4 genes in hippocampus cells as one of the tissues with high zinc density. Material and methods: First, the cell passage of the hippocampus cell line was performed, then the MTT assay test was performed for zinc oxide nanoparticles. In the next step, RNA extraction and CDNA synthesis were performed, and nanodrop spectrophotometer was used to ensure the purity of the RNA samples. Specific and appropriate primers of the desired genes were designed and synthesized. Then, changes in the expression of Znt1, Znt2, Znt3, and Znt4 genes were investigated using Real-Tim e RT-PCR. Results: Concentrations of 10 and 20 μg/mL of zinc oxide nanoparticles, significantly increased the expression of Znt1, Znt2, Znt3, and Znt4 genes in the hippocampus cell line of rat, while creating the lowest cytotoxicity. Conclusion: Zinc oxide nanoparticles can be investigated pharmacologically by increasing the expression of Znt1, Znt2, Znt3, and Znt4 genes in the treatment of zinc homeostasis disorders such as Alzheimer, depression, learning disabilities and stress.
Gower-Winter SD, Levenson CW. Zinc in the central nervous system: From molecules to behavior. Biofactors. 2012; 38(3): 186-93.
Takeda A, Tamano H, Ogawa T, Takada S, Ando M, Oku N, Watanabe M. Significance of serum glucocorticoid and chelatable zinc in depression and cognition in zinc deficiency. Behav Brain Res. 2012; 226(1): 259-64.
Dou X, Tian X, Zheng Y, Huang J, Shen Z, Li H & et al. Psychological stress induced hippocampus zinc dyshomeostasis and depression-like behavior in rats. Behav Brain Res. 2014; 273: 133-8.
Prasad AS. Impact of the discovery of human zinc deficiency on health. J Am Coll Nutr. 2009; 28: 257-65.
Song Y, Elias V, Wong CP, Scrimgeour AG, Ho E & et al. Zinc transporter expression profiles in the rat prostate following alterations in dietary zinc. Biometals. 2010; 23(1): 51-8.
Pfaender S, Föhr K, Lutz AK, Putz S, Achberger K, Linta L & et al. Cellular zinc homeostasis contributes to neuronal differentiation in human induced pluripotent stem Cells. Neural Plast. 2016; 3760702.
Chowanadisai W, Graham DM, Keen CL, Rucker RB, Messerli MA. A zinc transporter gene required for development of the nervous system. Commun Integr Biol. 2013; 6(6): e26207.
Gower-Winter SD, Corniola RS, Morgan Jr TJ, Levenson CW. Zinc deficiency regulates hippocampal gene expression and impairs neuronal differentiation. Nutr Neurosci. 2013; 16(4): 174-82.
Bin BH, Seo J, Kim ST. Function, structure, and transport aspects of ZIP and ZnT zinc transporters in immune cells. J Immunol Res. 2018; 9365747.
Downey AM, Hales BF, Robaire B. Zinc transport differs in rat spermatogenic cell types and is affected by treatment with cyclophosphamide. Biol Reprod. 2006; 95(1): 22.
Jobarteh ML, Mcardle HJ, Holtrop G, Sise EA, Prentice AM, Moor SE. mRNA levels of placental iron and zinc transporter genes are upregulated in gambian women with low iron and zinc status. J Nutr. 2017; 1401-9.
Lee S, Hennigar SR, Alam S, Nishida K, Kelleher SL. Essential role for zinc transporter
2 (ZnT2)-mediated zinc transport in mammary gland development and function during lactation. J Biol Chem. 2015; 290 (21): 13064-78.
Lee JY, Kim JS, Byun HR, Palmiter RD, Koh JY. Dependence of the histofluorescently reactive zinc pool on zinc transporter-3 in the normal brain. Brain Res. 2011; 1418: 12-22.
McCormick NH, Kelleher SL. ZnT4 provides zinc to zinc-dependent proteins in the
trans-Golgi network critical for cell function and Zn export in mammary epithelial cells. Am J Physiol Cell Physiol. 2012; 303 (3): 291-7.
Jiang J, Pi J, Cai J. The advancing of zinc oxide nanoparticles for biomedical applications. Bioinorg Chem Appl. 2011; 1062562.
Scherzad A, Meyer T, Kleinsasser N, Hackenberg S. Molecular mechanisms of zinc
oxide nanoparticle-induced genotoxicity short running title: genotoxicity of ZnO NPs. Materials. 2017; 10(12): 1427.
Chevallet M, Gallet B, Fuchs A, Jouneau PH, Um K, Mintz E. Metal homeostasis disruption and mitochondrial dysfunction in hepatocytes exposed to sub-toxic doses of zinc oxide nanoparticles. Nanoscale. 2016; 8(43): 18495-506.
Hanagata N, Xu M, Takemura T, Zhuang F. Cellular response to ZnO nanoparticle toxicity inferred from global gene expression profiles. Nano Biomed. 2010; 2(2): 153-69.
Zheng Y, Huang J, Tao L, Shen Z, Li H, Mo F & et al. Corticosterone increases intracellular Zn (2+) release in hippocampal HT-22 cells. Neurosci Lett. 2015; 588: 172-7.
Portbury SD, Adlard PA. Zinc signal in brain diseases. Int J Mol Sci. 2017; 18(2): 2506.
Torabi M, Kesmati M, Pourreza N, Varzi HN, Galehdari H. Neurobehavioral and biochemical modulation following administration of MgO and ZnO nanoparticles in the presence and absence of acute stress. Life Sci. 2018; 203: 72-82.
Wang C, Lu J, Zhou L, Li J, Xu J, Li W & et al. Effects of long-term exposure to zinc oxide nanoparticles on development, zinc metabolism and biodistribution of minerals (Zn, Fe, Cu, Mn) in mice. PLoS. 2016; 11(10): e0164434.
Lehvy AI, Horev G, Golan Y, Shammai Y, Assaraf YG. Alterations in ZnT1 expression and function lead to impaired intracellular zinc homeostasis in cancer. Cell Death Discov. 2019; 5: 144.
Chowanadisai W, Kelleher SL, Lönnerdal B. Zinc deficiency is associated with increased brain zinc import and LIV-1 expression and decreased ZnT-1 expression in neonatal rats. Nutr Neurosci. 2005; 135(5): 1002-7.
Liuzzi JP, Blanchard RK, Cousins RO. Differential regulation of zinc transporter 1, 2, and 4 mRNAexpression by dietary zinc in rats. J Nutr. 2001; 131:46-52.
Pfaffl MW, Windisch W. Influence of zinc deficiency on the mRNA expression of zinc transporters in adult rats. J Trace Elem Med Biol. 2003; 17(2): 97-106.
_||_Gower-Winter SD, Levenson CW. Zinc in the central nervous system: From molecules to behavior. Biofactors. 2012; 38(3): 186-93.
Takeda A, Tamano H, Ogawa T, Takada S, Ando M, Oku N, Watanabe M. Significance of serum glucocorticoid and chelatable zinc in depression and cognition in zinc deficiency. Behav Brain Res. 2012; 226(1): 259-64.
Dou X, Tian X, Zheng Y, Huang J, Shen Z, Li H & et al. Psychological stress induced hippocampus zinc dyshomeostasis and depression-like behavior in rats. Behav Brain Res. 2014; 273: 133-8.
Prasad AS. Impact of the discovery of human zinc deficiency on health. J Am Coll Nutr. 2009; 28: 257-65.
Song Y, Elias V, Wong CP, Scrimgeour AG, Ho E & et al. Zinc transporter expression profiles in the rat prostate following alterations in dietary zinc. Biometals. 2010; 23(1): 51-8.
Pfaender S, Föhr K, Lutz AK, Putz S, Achberger K, Linta L & et al. Cellular zinc homeostasis contributes to neuronal differentiation in human induced pluripotent stem Cells. Neural Plast. 2016; 3760702.
Chowanadisai W, Graham DM, Keen CL, Rucker RB, Messerli MA. A zinc transporter gene required for development of the nervous system. Commun Integr Biol. 2013; 6(6): e26207.
Gower-Winter SD, Corniola RS, Morgan Jr TJ, Levenson CW. Zinc deficiency regulates hippocampal gene expression and impairs neuronal differentiation. Nutr Neurosci. 2013; 16(4): 174-82.
Bin BH, Seo J, Kim ST. Function, structure, and transport aspects of ZIP and ZnT zinc transporters in immune cells. J Immunol Res. 2018; 9365747.
Downey AM, Hales BF, Robaire B. Zinc transport differs in rat spermatogenic cell types and is affected by treatment with cyclophosphamide. Biol Reprod. 2006; 95(1): 22.
Jobarteh ML, Mcardle HJ, Holtrop G, Sise EA, Prentice AM, Moor SE. mRNA levels of placental iron and zinc transporter genes are upregulated in gambian women with low iron and zinc status. J Nutr. 2017; 1401-9.
Lee S, Hennigar SR, Alam S, Nishida K, Kelleher SL. Essential role for zinc transporter
2 (ZnT2)-mediated zinc transport in mammary gland development and function during lactation. J Biol Chem. 2015; 290 (21): 13064-78.
Lee JY, Kim JS, Byun HR, Palmiter RD, Koh JY. Dependence of the histofluorescently reactive zinc pool on zinc transporter-3 in the normal brain. Brain Res. 2011; 1418: 12-22.
McCormick NH, Kelleher SL. ZnT4 provides zinc to zinc-dependent proteins in the
trans-Golgi network critical for cell function and Zn export in mammary epithelial cells. Am J Physiol Cell Physiol. 2012; 303 (3): 291-7.
Jiang J, Pi J, Cai J. The advancing of zinc oxide nanoparticles for biomedical applications. Bioinorg Chem Appl. 2011; 1062562.
Scherzad A, Meyer T, Kleinsasser N, Hackenberg S. Molecular mechanisms of zinc
oxide nanoparticle-induced genotoxicity short running title: genotoxicity of ZnO NPs. Materials. 2017; 10(12): 1427.
Chevallet M, Gallet B, Fuchs A, Jouneau PH, Um K, Mintz E. Metal homeostasis disruption and mitochondrial dysfunction in hepatocytes exposed to sub-toxic doses of zinc oxide nanoparticles. Nanoscale. 2016; 8(43): 18495-506.
Hanagata N, Xu M, Takemura T, Zhuang F. Cellular response to ZnO nanoparticle toxicity inferred from global gene expression profiles. Nano Biomed. 2010; 2(2): 153-69.
Zheng Y, Huang J, Tao L, Shen Z, Li H, Mo F & et al. Corticosterone increases intracellular Zn (2+) release in hippocampal HT-22 cells. Neurosci Lett. 2015; 588: 172-7.
Portbury SD, Adlard PA. Zinc signal in brain diseases. Int J Mol Sci. 2017; 18(2): 2506.
Torabi M, Kesmati M, Pourreza N, Varzi HN, Galehdari H. Neurobehavioral and biochemical modulation following administration of MgO and ZnO nanoparticles in the presence and absence of acute stress. Life Sci. 2018; 203: 72-82.
Wang C, Lu J, Zhou L, Li J, Xu J, Li W & et al. Effects of long-term exposure to zinc oxide nanoparticles on development, zinc metabolism and biodistribution of minerals (Zn, Fe, Cu, Mn) in mice. PLoS. 2016; 11(10): e0164434.
Lehvy AI, Horev G, Golan Y, Shammai Y, Assaraf YG. Alterations in ZnT1 expression and function lead to impaired intracellular zinc homeostasis in cancer. Cell Death Discov. 2019; 5: 144.
Chowanadisai W, Kelleher SL, Lönnerdal B. Zinc deficiency is associated with increased brain zinc import and LIV-1 expression and decreased ZnT-1 expression in neonatal rats. Nutr Neurosci. 2005; 135(5): 1002-7.
Liuzzi JP, Blanchard RK, Cousins RO. Differential regulation of zinc transporter 1, 2, and 4 mRNAexpression by dietary zinc in rats. J Nutr. 2001; 131:46-52.
Pfaffl MW, Windisch W. Influence of zinc deficiency on the mRNA expression of zinc transporters in adult rats. J Trace Elem Med Biol. 2003; 17(2): 97-106.