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Antioxidant, cytokine, Encephalitozoon intestinalis, oxidative stress, U937


This in vitro study was performed to investigate the changes in nitric oxide (NO), malondialdehyde (MDA), total antioxidant capacity (TAC), tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) levels in human monocytic (U937) cells infected with Encephalitozoon intestinalis. E. intestinalis was first cultured in the African green monkey renal epithelial (Vero) cells to obtain sufficient amount of parasite for the study. U937 human macrophage cell line and E. intestinalis 50506 (ATCC) strain were used in the study. U937 macrophages were infected with 5x106 spores after stimulation with Phorbol 12-Myristate 13-Acetate (PMA) and uninfected U937 cells were used as control. Culture media were collected at the 6th, 12th, and 24th hour after infection to determine NO, MDA, TAC, TNF-α and IL-10 levels. NO levels significantly increased at the 6th, 12th and 24th hour. There was no significant difference in MDA and TAC levels at the 6th and 12th hour, but a significant increase in MDA, and a significant decrease in TAC was observed at the 24th hour. TNF-α levels did not differ in all sampling times, and IL-10 level decreased only at the 6th hour. In conclusion, E. intestinalis caused oxidative stress by increasing the levels of oxidants (NO and MDA) and by suppressing TAC level without no significant changes in cytokine levels in U937 cell line.


Aseeja, P., Shaikh, Y., Bajpai, A., Sirsikar, P., Kalra, S.K. (2021): Advancement in our understanding of immune response against Encephalitozoon infection. Parasite Immunol 43.

Han, B., Pan, G., Weiss, L.M. (2021): Microsporidiosis in Humans. Clin Microbiol Rev 34(4): e0001020.

Curry, A. (2005): Microsporidiosis, In: Cox, F.E.G., Wakelin, D., Gillespie, S.H., Despommier, D.D. (eds.) Topley and Wilson’s Microbiology and Microbial Infections; Parasitology. ASM Press. Washington. pp. 529-555.

Cetinkaya, Ü., Caner, A. (2020): Prevalence of microsporidiosis in different hosts in turkey: a meta-analysis. Turkiye Parazitol Derg 44: 232-238.

Han, B., Takvorian, P.M., Weiss, L.M. (2020): Invasion of Host Cells by Microsporidia. Front Microbiol 11: 172.

Moretto, M.M., Khan, I.A. (2022): Immune Response to Microsporidia. Exp Suppl 114: 373-388.

Canton, M., Sánchez-Rodríguez, R., Spera, I., Venegas, F.C., Favia, M., Viola, A., Castegna, A. (2021): Reactive oxygen species in macrophages: sources and targets. Front Immunol 12: 734229.

Polat, M., Kiliç, E., Yazar, S., Çetinkaya. Ü. (2012): Toxoplasma gondii pozitif bireylerde ileri protein oksidasyon ürünü düzeylerinin değerlendirilmesi. ERÜ Sağlık Bil Derg 21: 200-204.

Abu-Dieyeh, Z. (2006): Effect of high temperature per se on growth performance of broilers. Int J Poult Sci 5: 19-21.

Sindhu, R.K., Koo, J., Roberts, K.C., Vaziri, N.D. (2004): Dysregulation of hepatic superoxide dismutase, catalase to insulin to insulin and antioxidant therapies. Clin Exp Hypertens Journal 26: 43-53.

Venditti, P., Daniele, M.C., Masullo, P., Di Meo, S. (1999): Antioxidant-sensitive triiodothyronine effects on characteristics of rat liver mitochondrial population. Cell Physiol Biochem 9: 38-52, 1999.

Feng, Z., Hu, W., Marnett, L.J., Tang, M.S. (2006): Malondialdehyde, a major endogenous lipid peroxidation product, sensitizes human cells to UV and BPDE induced killing and mutagenesis through inhibition of nucleotide excision repair. Mutat Res 601: 125-136.

Deger, Y., Ertekin, A., Değer, S., Mert, H. (2008): Lipid peroxidation and antioxidant potential of sheep liver infected naturally with distomatosis. Turkiye Parazitol Derg 32: 23-26.

Sussmann, R.A.C., Fotoran, W.L., Kimura, E.A., Katzin, A.M. (2017): Plasmodium falciparum uses vitamin E to avoid oxidative stress. Parasit Vectors 10: 461.

Sasani, M., Nabavi, R., Hajinezhad, M., Hasanein, P. (2018): Oxidative stress and hepatic injury induced in mice fed a Sarcocystis hirsuta cyst extract. J Vet Sci 19: 500-504.

Bredt, D.S., Snyder, S.H. (1994): Nitric oxide: a physiologic messenger molecule. Annu Rev Biochem 63: 175-195.

Suzuki, Y. (1999): Genes, cells and cytokines in resistance against development of toxoplasmic encephalitis. Imbio 201: 255-271.

Viviani, B., Bartesaghi, S., Corsini, E., Galli, C.L., Marinovich, M. (2004): Cytokines role in neurodegenerative events. Toxicol Lett 149: 85-89.

Yilmaz, O., Taskiran, D., Aydar, S. (2004): Cytotoxicity in cytokine stimulated astrocyte cultures: role of IL-6 and nitric oxide. Neurosci Res Commun 34: 82-91.

Asadullah, K., Sterry, W., Volk, H.D. (2003): Interleukin-10 therapy–review of a new approach. Pharmacol 55: 241-269.

Franzen, C. (2005): How do microsporidia invade cells? Folia Parasitol (Praha) 52: 36-40.

Franzen, C., Hartmann, P., Salzberger, B. (2005): Cytokine and nitric oxide responses of monocyte-derived human macrophages to microsporidian spores. Exp Parasitol 109: 1-6.

Chandramathi, S., Suresh, K., Anita, Z.B., Kuppusamy, U.R. (2009): Elevated levels of urinary hydrogen peroxide, advanced oxidative protein product (AOPP) and malondialdehyde in humans infected with intestinal parasites. Parasitology 136: 359-363.

Catterall, J.R., Sharma, S.D., Remington, J.S. (1986): Oxygen-independent killing by alveolar macrophages. J Exp Med 163: 1113-1131.

Chang, H.R., Pechere, J.C. (1989): Macrophage oxidative metabolism and intracellular Toxoplasma gondii. Microb Pathog 7: 37-44.

Didier, E.S., Sokolova, Y.Y., Alvarez, X., Bowers, L.C. (2009): Encephalitozoon cuniculi (Microsporidia) suppresses apoptosis in human macrophages. J Immunol 182: 133.

He, Q., Leitch, G.J., Visvesvara, G.S., Wallace, S. (1996): Effects of nifedipine, metronidazole, and nitric oxide donors on spore germination and cell culture infection of the microsporidia Encephalitozoon hellem and Encephalitozoon intestinalis. Antimicrob Agents Chemoth er 40: 179-185.

Rosselli, M., Dubey, R.K., Rosselli, M.A., Macas, E., Fink, D., Lauper, U., Keller, P.J., Imthurn, B. (1996): Identification of nitric oxide synthase in human and bovine oviduct. Mol Hum Reprod 2: 607-612.

Forstermann, U., Schmidt, H.H.W., Pollock, J.S., Sheng, H., Mitchell, J.A., Warner, T.D., Nakane, M., Murad, F. (1991): Isoforms of nitric oxide synthase characterization and purification from different cell types. Biochem Pharmacol 42: 1849-1857.

Forstermann, U., Boissel, J.P., Kleinert, H. (1998): Expressional control of the constitutive isoforms of nitric oxide synthase (NOS I and NOS III). FASEB J 12: 773-790.

Nathan, C., Xie, Q.W. (1994): Regulation of biosynthesis of nitric oxide. J Biol Chem 269: 13725-13728.

Foley, E., O'farrell, P.H. (2003): Nitric oxide contributes to induction of innate immune responses to gram-negative bacteria in Drosophila. Genes Dev 17: 115-125.

Flores-Villegas, A.L., Salazar-Schettino, P.M., Córdoba-Aguilar, A., Gutiérrez-Cabrera, A.E., Rojas-Wastavino, G.E., Bucio-Torres, M.I., Cabrera-Bravo, M. (2015): Immune defence mechanisms of triatomines against bacteria, viruses, fungi and parasites. Bull Entomol Res 105: 523-532.

Abd Ellah, M.R. (2013): Involvement of free radicals in parasitic infestations. J Appl Behav Anal 41: 69-76.

Smith, A.R., Visioli, F., Hagen, T.M. (2002): Vitamin C matters: Increased oxidative stress in culture human aortic endothelial cells without supplemental ascorbic acid. FASEB J 16: 1102 1104.

Halliwell, B. (2014): Cell culture, oxidative stress, and antioxidants: avoiding pitfalls. Biomed J 37: 99-105.

Launois, P., Tacchini-Cottier, F., Parra-Lopez, C., Louis, J.A. (1998): Cytokines in parasitic diseases: the example of cutaneous leishmaniasis. Intern Rev Immunol 17: 157-180.

Nevárez-Garza, A.M., Castillo-Velázquez, U., Soto-Domínguez, A., Montes-De-Oca-Luna, R., Zamora-Ávila, D.E., Wong-González, A., Rodríguez-Tovar, L.E. (2018): Quantitative analysis of TNF-α, IL-4, and IL-10 expression, nitric oxide response, and apoptosis in Encephalitozoon cuniculi-infected rabbits. Dev Comp Immunol 81: 235-243.

Rodríguez-Tovar, L.E., Castillo-Velázquez, U., Arce-Mendoza, A.Y., Nevárez-Garza, A.M., Zarate-Ramos, J.J., Hernández-Vidal, G., Rodríguez-Ramírez, H.G., Trejo-Chávez, A. (2016): Interferon γ and interleukin 10 responses in immunocompetent and immunosuppressed New Zealand White rabbits naturally infected with Encephalitozoon cuniculi. Dev Comp Immunol 62: 82-88.




How to Cite

Şentürk, M., Çetinkaya, Ülfet, Eren, M., & Charyyeva , A. (2023). THE EFFECT OF ENCEPHALITOZOON INTESTINALIS ON OXIDATIVE STRESS AND CYTOKINE LEVELS IN U937 CELLS: AN IN VITRO STUDY. Journal of Applied Biological Sciences, 17(1), 155–162. Retrieved from




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