The alterations in cytokine mRNA expressions and productions by fluoride in a murine macrophage cell line evaluated by real-time PCR and ELISA
We hypothesized that macrophages have a role in the alterations in bone metabolism induced by fluoride via cytokines. In this study, interleukin-10 (IL-10) and IL-12 in addition to tumor necrosis factor α (TNFα) and IL-1β were evaluated for their mRNA expressions in J774.1 cells by quantitative real...
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| Published in | Biomedical Research on Trace Elements Vol. 22; no. 1; pp. 27 - 33 |
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| Main Authors | , , , , , , , |
| Format | Journal Article |
| Language | English Japanese |
| Published |
Osaka
Japan Society for Biomedical Research on Trace Elements
01.01.2011
Japan Science and Technology Agency |
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| Online Access | Get full text |
| ISSN | 0916-717X 1880-1404 |
| DOI | 10.11299/brte.22.27 |
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| Abstract | We hypothesized that macrophages have a role in the alterations in bone metabolism induced by fluoride via cytokines. In this study, interleukin-10 (IL-10) and IL-12 in addition to tumor necrosis factor α (TNFα) and IL-1β were evaluated for their mRNA expressions in J774.1 cells by quantitative real-time PCR and the protein levels in the supernatant from the cell culture by ELISA at 0, 100, 300, and 1000 µM fluoride. At 18 hours after incubation, J774.1 cells were activated by lipopolysaccharide. At 6 hours after the activation, RNAs of the cells were sampled. The mRNA expressions for TNFα, IL-1β, IL-10, and IL-12p40 were analyzed by real-time PCR. The supernatant from the cell culture were sampled at 24 hours after the activation and determined for these cytokine levels by ELISA. The cell viability in the 1000 µM group at 6 hours after the activation was significantly lower than that in the control. The mRNA expressions of TNFα, IL-1β and IL-10 in the 1000 µM group were significantly higher than those in the control. Although it did not reach the significant level, the mRNA expression of IL-12p40 in the 1000 µM group was elevated more than that in the control. The mean values of concentrations of TNFα and IL-1β in the supernatant in the 1000 µM group were significantly lower than those in the control. While, there was no significant difference for the concentration of IL-10 in the supernatant among the groups. For IL-12p40, the mean value of the concentration in the 1000 µM group was significantly higher than that in the control. The effects of fluoride on the cytokines produced by macrophages were not common for all types of cytokines. In conclusion, the effects of fluoride on the immune system may vary via alterations in cytokine productions by macrophages, resulting in the alterations in bone metabolism. |
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| AbstractList | We hypothesized that macrophages have a role in the alterations in bone metabolism induced by fluoride via cytokines. In this study, interleukin-10 (IL-10) and IL-12 in addition to tumor necrosis factor α (TNFα) and IL-1β were evaluated for their mRNA expressions in J774.1 cells by quantitative real-time PCR and the protein levels in the supernatant from the cell culture by ELISA at 0, 100, 300, and 1000 µM fluoride. At 18 hours after incubation, J774.1 cells were activated by lipopolysaccharide. At 6 hours after the activation, RNAs of the cells were sampled. The mRNA expressions for TNFα, IL-1β, IL-10, and IL-12p40 were analyzed by real-time PCR. The supernatant from the cell culture were sampled at 24 hours after the activation and determined for these cytokine levels by ELISA. The cell viability in the 1000 µM group at 6 hours after the activation was significantly lower than that in the control. The mRNA expressions of TNFα, IL-1β and IL-10 in the 1000 µM group were significantly higher than those in the control. Although it did not reach the significant level, the mRNA expression of IL-12p40 in the 1000 µM group was elevated more than that in the control. The mean values of concentrations of TNFα and IL-1β in the supernatant in the 1000 µM group were significantly lower than those in the control. While, there was no significant difference for the concentration of IL-10 in the supernatant among the groups. For IL-12p40, the mean value of the concentration in the 1000 µM group was significantly higher than that in the control. The effects of fluoride on the cytokines produced by macrophages were not common for all types of cytokines. In conclusion, the effects of fluoride on the immune system may vary via alterations in cytokine productions by macrophages, resulting in the alterations in bone metabolism. We hypothesized that macrophages have a role in the alterations in bone metabolism induced by fluoride via cytokines. In this study, interleukin-10 (IL-10) and IL-12 in addition to tumor necrosis factor α (TNFα) and IL-1β were evaluated for their mRNA expressions in J774.1 cells by quantitative real-time PCR and the protein levels in the supernatant from the cell culture by ELISA at 0, 100, 300, and 1000 μM fluoride. At 18 hours after incubation, J774.1 cells were activated by lipopolysaccharide. At 6 hours after the activation, RNAs of the cells were sampled. The mRNA expressions for TNFα, IL-1β, IL-10, and IL-12p40 were analyzed by real-time PCR. The supernatant from the cell culture were sampled at 24 hours after the activation and determined for these cytokine levels by ELISA. The cell viability in the 1000 μM group at 6 hours after the activation was significantly lower than that in the control. The mRNA expressions of TNFα, IL-1β and IL-10 in the 1000μM group were significantly higher than those in the control. Although it did not reach the significant level, the mRNA expression of IL-12p40 in the 1000 μM group was elevated more than that in the control. The mean values of concentrations of TNFα and IL-1β in the supernatant in the 1000 μM group were significantly lower than those in the control. While, there was no significant difference for the concentration of IL-10 in the supernatant among the groups. For IL-12p40, the mean value of the concentration in the 1000 μM group was significantly higher than that in the control. The effects of fluoride on the cytokines produced by macrophages were not common for all types of cytokines. In conclusion, the effects of fluoride on the immune system may vary via alterations in cytokine productions by macrophages, resulting in the alterations in bone metabolism. |
| Author | Ito, Kyoko Tsunoda, Masashi Aizawa, Yoshiharu Chen, Xia-Fen Tsunoda, Humio Inoue, Yoko Katagiri, Hiroshi Hosokawa, Mayuko |
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| References | 6) Blair HC, Sidonio RF, Friedberg RC, Khan NN, Dong SS: Proteinase expression during differentiation of human osteoclasts in vitro. J Cell Biochem 78: 627-637, 2000. 15) Tsunoda M, Yoshida T, Tsuji M, Zhan Y, Sugaya C, Inoue Y, Miki T, Kudo Y, Satoh T, Aizawa Y: The effects of dibutyltin (DBT) dichloride on the viability and the productions of tumor necrosis factor α and interleukin-12 in murine macrophage cell line, J774.1. Biomed Res Trace Elements 19: 67-71, 2008. 18) Tamer MN, Kale Koroglu B, Arslan C, Akdogan M, Koroglu M, Cam H, Yildiz M: Osteosclerosis due to endemic fluorosis. Sci Total Environ 373: 43-48, 2007. 11) Nakano K, Tsunoda M, Konno N: Tributylin (TBT) increases TNFα mRNA expression and induces apoptosis in the murine macrophage cell line in vitro. Environ Health Prev Med 9: 266-271, 2004. 17) Kono K: Health effects of fluoride and its compounds. Jpn J Hyg 49: 852-860, 1994. (in Japanese 10) Sorgdrager B, Pal TM, Loff de AJ, Dubois AE, Monchy de JG: Occupational asthma in aluminum potroom workers related to pre-employment eosinophil count. Eur Respir J 8: 1520-1524, 1995. 13) Tsunoda M, Sharma RP: Modulation of tumor necrosis factor a expression in mouse brain after exposure to aluminum in drinking water. Arch Toxicol 73: 419-426, 1999. 8) Hosokawa M, Sugaya C, Inoue Y, Tsunoda M, Aizawa Y: Cytotoxicity and inhibition of the production of tumor necrosis factor α and interleukin-1β induced macrophage cell line. Fluoride 42: 188-197, 2009. 14) Reiner SL, Zheng S, Corry DB, Locksley RM: Constructing polycompetitor cDNAs for quantitative PCR. J Immunol Methods 165: 37-46, 1993. 1) Qureshi R, House R, Uhlig E, Genesov L, Holness DL: Hydrofluoric acid burn: case report. J Can Assoc Emerg Physician 4: 292-295, 2002. 5) Matsuo S, Kiyomiya K, Kurebe M: Mechanism of toxic action of fluoride in dental fluorosis: whether trimeric G proteins participate in the disturbance of intracellular transport of secretory ameloblast exposed to fluoride. Arch Toxicol 72: 798-806, 1998. 9) Yata J: Cytokines and their roles. Yata J: Textbook of Immunology for Medical Students and Physicians. 11th edition. Chugai Igakusya, Tokyo, 2009, pp.306-361. (in Japanese 19) Itai, K. Micro-determination of fluoride in biological samples by pyrohydrolysis and flow-injection analysis using a fluoride ion-selective electrode. Jpn J Hyg 45: 1061-1073, 1991. (in Japanese 3) Zhang B, Hong M, Zhang B, Zhang XL, Zhao YS: Fluorine distribution in aquatic environment and its health effect in the Western Region of the Songnen Plain, Northeast China. Environ Monit Assess 133: 379-386, 2007. 7) Teitelbaum SL: Bone resorption by osteoclasts. Science 289: 1504-1508, 2000. 4) Choubisa SL, Sompura K, Bhatt SK, Choubisa DK, Pandya H, Joshi SC, Choubisa L: Prevalence of fluorosis in some villages of Dungarpur district of Rajasthan. Indian J Environ Health 38: 119-126: 1996. 2) Dote T, Kono K: An acute lethal case of exposure during a washing down operation of a hydrogen fluoride liquefying tank. Jpn J Occup Med Traumatol 52: 189-192, 2004. (in Japanese 12) Tsunoda M, Yamamoto M, Ito K, Inoue Y, Miki T, Kudo Y, Satoh T, Aizawa Y: Dibutyltin (DBT) dichloride inhibits cytokine productions in murine macrophage cell line, J774.1. Biomed Res Trace Elements 17: 417-422, 2006. 16) Ando M, Tadano M, Yamamoto S, Tamura K, Asanuma S, Watanabe T, Kondo T, Sakurai S, Ji R, Liang C, Chen X, Hong Z, Cao S: Health effects of fluoride pollution caused by coal burning. Sci Total 271: 107-116, 2001. |
| References_xml | – reference: 10) Sorgdrager B, Pal TM, Loff de AJ, Dubois AE, Monchy de JG: Occupational asthma in aluminum potroom workers related to pre-employment eosinophil count. Eur Respir J 8: 1520-1524, 1995. – reference: 14) Reiner SL, Zheng S, Corry DB, Locksley RM: Constructing polycompetitor cDNAs for quantitative PCR. J Immunol Methods 165: 37-46, 1993. – reference: 16) Ando M, Tadano M, Yamamoto S, Tamura K, Asanuma S, Watanabe T, Kondo T, Sakurai S, Ji R, Liang C, Chen X, Hong Z, Cao S: Health effects of fluoride pollution caused by coal burning. Sci Total 271: 107-116, 2001. – reference: 17) Kono K: Health effects of fluoride and its compounds. Jpn J Hyg 49: 852-860, 1994. (in Japanese) – reference: 8) Hosokawa M, Sugaya C, Inoue Y, Tsunoda M, Aizawa Y: Cytotoxicity and inhibition of the production of tumor necrosis factor α and interleukin-1β induced macrophage cell line. Fluoride 42: 188-197, 2009. – reference: 18) Tamer MN, Kale Koroglu B, Arslan C, Akdogan M, Koroglu M, Cam H, Yildiz M: Osteosclerosis due to endemic fluorosis. Sci Total Environ 373: 43-48, 2007. – reference: 1) Qureshi R, House R, Uhlig E, Genesov L, Holness DL: Hydrofluoric acid burn: case report. J Can Assoc Emerg Physician 4: 292-295, 2002. – reference: 7) Teitelbaum SL: Bone resorption by osteoclasts. Science 289: 1504-1508, 2000. – reference: 15) Tsunoda M, Yoshida T, Tsuji M, Zhan Y, Sugaya C, Inoue Y, Miki T, Kudo Y, Satoh T, Aizawa Y: The effects of dibutyltin (DBT) dichloride on the viability and the productions of tumor necrosis factor α and interleukin-12 in murine macrophage cell line, J774.1. Biomed Res Trace Elements 19: 67-71, 2008. – reference: 6) Blair HC, Sidonio RF, Friedberg RC, Khan NN, Dong SS: Proteinase expression during differentiation of human osteoclasts in vitro. J Cell Biochem 78: 627-637, 2000. – reference: 2) Dote T, Kono K: An acute lethal case of exposure during a washing down operation of a hydrogen fluoride liquefying tank. Jpn J Occup Med Traumatol 52: 189-192, 2004. (in Japanese) – reference: 5) Matsuo S, Kiyomiya K, Kurebe M: Mechanism of toxic action of fluoride in dental fluorosis: whether trimeric G proteins participate in the disturbance of intracellular transport of secretory ameloblast exposed to fluoride. Arch Toxicol 72: 798-806, 1998. – reference: 12) Tsunoda M, Yamamoto M, Ito K, Inoue Y, Miki T, Kudo Y, Satoh T, Aizawa Y: Dibutyltin (DBT) dichloride inhibits cytokine productions in murine macrophage cell line, J774.1. Biomed Res Trace Elements 17: 417-422, 2006. – reference: 13) Tsunoda M, Sharma RP: Modulation of tumor necrosis factor a expression in mouse brain after exposure to aluminum in drinking water. Arch Toxicol 73: 419-426, 1999. – reference: 4) Choubisa SL, Sompura K, Bhatt SK, Choubisa DK, Pandya H, Joshi SC, Choubisa L: Prevalence of fluorosis in some villages of Dungarpur district of Rajasthan. Indian J Environ Health 38: 119-126: 1996. – reference: 11) Nakano K, Tsunoda M, Konno N: Tributylin (TBT) increases TNFα mRNA expression and induces apoptosis in the murine macrophage cell line in vitro. Environ Health Prev Med 9: 266-271, 2004. – reference: 19) Itai, K. Micro-determination of fluoride in biological samples by pyrohydrolysis and flow-injection analysis using a fluoride ion-selective electrode. Jpn J Hyg 45: 1061-1073, 1991. (in Japanese) – reference: 3) Zhang B, Hong M, Zhang B, Zhang XL, Zhao YS: Fluorine distribution in aquatic environment and its health effect in the Western Region of the Songnen Plain, Northeast China. Environ Monit Assess 133: 379-386, 2007. – reference: 9) Yata J: Cytokines and their roles. Yata J: Textbook of Immunology for Medical Students and Physicians. 11th edition. Chugai Igakusya, Tokyo, 2009, pp.306-361. (in Japanese) |
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| Title | The alterations in cytokine mRNA expressions and productions by fluoride in a murine macrophage cell line evaluated by real-time PCR and ELISA |
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