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|標題:||Inhibitory effect of nano silicate platelets on the cytotoxicity of fumonisin B1 to mouse embryos
|關鍵字:||伏馬鐮孢毒素B1;小鼠胚胎;黴菌毒素吸附劑;奈米矽片;神經管缺陷;fumonisin B1 (FB1);mouse embryo;mycotoxins adsorbing agents;nano silicate platelets (NSP);neural tube defect||引用:||林秀橤。2009。影響Fusarium proliferatum產生fumonisin B1之因子探討。國立中興大學植物病理學系碩士論文。 鄭閔謙。2009。台灣畜產與水產飼糧中黴菌數與黴菌毒素含量之調查。國立中興大學動物科學系碩士論文。 Alberts, J. F., W. C. Gelderblom, P. G. Thiel, W. F. Marasas, D. J. Van Schalkwyk, and Y. Behrend. 1990. Effects of temperature and incubation period on production of fumonisin B1 by Fusarium moniliforme. Appl. Environ. Microbiol. 56:1729-1733. Alexandre, M., and P. Dubois. 2000. Polymer-layered silicate nanocomposites: preparation, properties and uses of a new class of materials. Mater. Sci. Eng. 28:1-63. Allende, M. L., J. L. Dreier, S. Mandala, and R. L. Proia. 2004. Expression of the sphingosine-1-phosphate receptor, S1P1, on T-cells controls thymic emigration. J. Biol. Chem. 279:15396-15401. Avantaggiato, G., M. Solfrizzo, and A. Visconti. 2005. 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目前已鑑定出300多種黴菌毒素，其中黃麴毒素（alfatoxin）、嘔吐毒素（deoxynivalenol, DON）、赭麴毒素（ochratoxin, OCH）、玉米毒素（F2毒素）（zearalenone, ZEA）與伏馬鐮孢毒素（fumonisins）等5種為飼料中常見之黴菌毒素。其中伏馬鐮孢毒素B1（fumonisin B1, FB1）為Fusarium moniliforme產生之黴菌毒素，直到1988年才被萃取出。飼料中常額外添加黴菌毒素吸附劑（mycotoxin adsorbing agents），解決黴菌毒素汙染之問題。Aluminosilicates為最大宗之黴菌毒素吸附劑，如bentonites、montmorillonites（MMT）、zeolites與hydrated sodium
calcium aluminosilicates（HSCAS）等。本試驗所用之奈米矽片（nano silicate platelets, NSP）係由MMT脫層而來。擁有極佳之非特異性結合能力，因此期能添加於動物飼料中吸附FB1。由於尚未有文獻指出奈米矽片對胚胎發育之影響，試驗先以體外與體內試驗探討奈米矽片對小鼠胚胎發育之毒性。結果顯示，奈米矽片對小鼠著床前胚之體外發育與品質並無任何影響。此外，灌食奈米矽片之母鼠其子代亦未發現任何異常。於吸附FB1能力方面，奈米矽片對FB1之體外吸附效率並不如預期，然而由血液中低濃度之FB1、減少神經管缺陷（neural tube defect, NTD）發生率、增進胎兒體重與提高胎兒longevity assurance homolog 5（LASS5）基因表現量之結果可知，奈米矽片於體內吸附FB1之能力相當高。依上述結果，動物飼料中添加奈米矽片以減少FB1對動物之毒害，應為可行之方法。此外，FB1引起NTD之詳細機制尚未完全明瞭，推測任何引起NTD之途徑皆有可能為FB1之作用機制。懷孕母鼠灌食FB1後分析肝臟、子宮、胎兒，以及胎盤中與NTD發生相關之sphingosine kinase 1（Sphk1）、sphingosine kinase 2（Sphk2）、sphingosine-1-phosphate（Sgpl1）、Sphingosine-1-phosphate lyase（Sgpp1）、N-acylsphingosine amidohydrolase 1（Asah1）、sphingomyelin phosphodiesterase 1（Smpd1）、betaine homocysteine methyltransferase（Bhmt）與Serine hydroxymethyltransferase 1（Shmt1）基因表現，發現神經鞘脂相關基因表現有異常現象。其中子宮之基因表現增幅程度最大，其餘組織之基因表現多呈現減少現象。然而，此結果僅限於基因層面，神經鞘脂本身之濃度是否隨基因表現改變而有所變化，仍需進一步探討。
More than 300 kinds of mycotoxins had been discovered, in which alfatoxin, deoxynivalenol (DON), ochratoxin (OCH), zearalenone (ZEA), and fumonisins are five major mycotoxins in animal feed. Fumonisin B1 (FB1) is a mycotoxin produced by Fusarium moniliforme. Supplementation of mycotoxin adsorbing agents is one of the solutions to prevent adsorption of high contamination of mycotoxins in feed by gastrointestinal tract. Aluminosilicates are the largest groups of mycotoxin adsorbing agents and commercially used, including bentonites, montmorillonites (MMT), zeolites, and hydrated sodium calcium aluminosilicates (HSCAS). In present study, nano silicate platelets (NSP) exfoliated from montmorillonite (MMT) with a high non-specific binding capacity might act as an additive in animal feed for adsorbing mycotoxin of FB1. However, the effect of NSP on the development of embryos is not yet revealed in literature. The toxicity of NSP on the development of mouse embryos was examined by both of in vitro and in vivo approaches. The results showed that NSP did not disturb the development and the quality of intact pre-implantation mouse embryos. No abnormalities for the newborn mice reproduced from the female fed with NSP were found. The NSP had an unexpected low adsorbing efficiency in vitro but a high adsorbing ability on FB1 in vivo, evidenced by a low conentration of FB1 in blood, reduced incidence of neural tube defect (NTD), increased weight of the fetus, and high gene expression of LASS5 (longevity assurance homolog 5). Based on these findings, we concluded that feed supplemented with NSP is a feasible way for reducing the toxicity of FB1 to animals. In addition, the mechanism of NTD caused by FB1 was not fully understood yet. We suspected that perturbation of sphingolipid, choline, folate, and methionine pathway by FB1 induced cytotoxicity leading to NTD. These related gene expresions, including sphingosine kinase 1 (Sphk1), sphingosine kinase 2 (Sphk2), sphingosine-1-phosphate (Sgpl1), Sphingosine-1-phosphate lyase (Sgpp1), N-acylsphingosine amidohydrolase 1 (Asah1), sphingomyelin phosphodiesterase 1 (Smpd1), betaine homocysteine methyltransferase (Bhmt), and Serine hydroxymethyltransferase 1 (Shmt1) in the liver, uterus, fetus, and placenta derived from pregnant FB1-treated mice, were found with abnormal expression pattern. The sphingolipid related gene expressions in uterus were dramatically increased, but that in other tissues were reduced mostly by FB1 treatment. These results were only based on the genetic level. Further studies should focus on the in situ concentration of sphingolipids to examine their change with different gene expressions.
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