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作为火鸡(吐绶鸡)硒状态和硒需求生物标志物的硒蛋白转录水平和酶活性

Selenoprotein Transcript Level and Enzyme Activity as Biomarkers for Selenium Status and Selenium Requirements in the Turkey (Meleagris gallopavo).

作者信息

Taylor Rachel M, Sunde Roger A

机构信息

Department of Nutritional Sciences, University of Wisconsin, Madison, Wisconsin, United States of America.

出版信息

PLoS One. 2016 Mar 23;11(3):e0151665. doi: 10.1371/journal.pone.0151665. eCollection 2016.

Abstract

The current National Research Council (NRC) selenium (Se) requirement for the turkey is 0.2 μg Se/g diet. The sequencing of the turkey selenoproteome offers additional molecular biomarkers for assessment of Se status. To determine dietary Se requirements using selenoprotein transcript levels and enzyme activities, day-old male turkey poults were fed a Se-deficient diet supplemented with graded levels of Se (0, 0.025, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.75, 1.0 μg Se/g diet) as selenite, and 12.5X the vitamin E requirement. Poults fed less than 0.05 μg Se/g diet had a significantly reduced rate of growth, indicating the Se requirement for growth in young male poults is 0.05 μg Se/g diet. Se deficiency decreased plasma GPX3 (glutathione peroxidase), liver GPX1, and liver GPX4 activities to 2, 3, and 7%, respectively, of Se-adequate levels. Increasing Se supplementation resulted in well-defined plateaus for all blood, liver and gizzard enzyme activities and mRNA levels, showing that these selenoprotein biomarkers could not be used as biomarkers for supernutritional-Se status. Using selenoenzyme activity, minimum Se requirements based on red blood cell GPX1, plasma GPX3, and pancreas and liver GPX1 activities were 0.29-0.33 μg Se/g diet. qPCR analyses using all 10 dietary Se treatments for all 24 selenoprotein transcripts (plus SEPHS1) in liver, gizzard, and pancreas found that only 4, 4, and 3 transcripts, respectively, were significantly down-regulated by Se deficiency and could be used as Se biomarkers. Only GPX3 and SELH mRNA were down regulated in all 3 tissues. For these transcripts, minimum Se requirements were 0.07-0.09 μg Se/g for liver, 0.06-0.15 μg Se/g for gizzard, and 0.13-0.18 μg Se/g for pancreas, all less than enzyme-based requirements. Panels based on multiple Se-regulated transcripts were effective in identifying Se deficiency. These results show that the NRC turkey dietary Se requirement should be raised to 0.3 μg Se/g diet.

摘要

美国国家研究委员会(NRC)目前规定的火鸡硒(Se)需求量为每克日粮0.2微克硒。火鸡硒蛋白组的测序为评估硒状态提供了额外的分子生物标志物。为了利用硒蛋白转录水平和酶活性来确定日粮硒需求量,给1日龄雄性火鸡雏鸡饲喂一种缺硒日粮,并添加不同水平的硒(0、0.025、0.05、0.1、0.2、0.3、0.4、0.5、0.75、1.0微克硒/克日粮)作为亚硒酸盐,以及12.5倍的维生素E需求量。饲喂低于0.05微克硒/克日粮的雏鸡生长速率显著降低,这表明幼龄雄性火鸡雏鸡生长所需的硒量为0.05微克硒/克日粮。缺硒使血浆谷胱甘肽过氧化物酶3(GPX3)、肝脏谷胱甘肽过氧化物酶1(GPX1)和肝脏谷胱甘肽过氧化物酶4(GPX4)的活性分别降至硒充足水平的2%、3%和7%。增加硒的添加量会使所有血液、肝脏和肌胃酶活性以及mRNA水平出现明确的平台期,这表明这些硒蛋白生物标志物不能用作超营养硒状态的生物标志物。根据红细胞GPX1、血浆GPX3以及胰腺和肝脏GPX1的活性,利用硒酶活性确定的最低硒需求量为0.29 - 0.33微克硒/克日粮。对肝脏、肌胃和胰腺中所有24种硒蛋白转录本(加上SEP1HS)采用所有10种日粮硒处理进行qPCR分析发现,缺硒分别仅使4种、4种和3种转录本显著下调,可作为硒生物标志物。只有GPX3和SELH mRNA在所有3种组织中均下调。对于这些转录本,肝脏的最低硒需求量为0.07 - 0.09微克硒/克,肌胃为0.06 - 0.15微克硒/克,胰腺为0.13 - 0.18微克硒/克,均低于基于酶的需求量。基于多种硒调节转录本的组合有效地识别了缺硒情况。这些结果表明,NRC规定的火鸡日粮硒需求量应提高到0.3微克硒/克日粮。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/56ce/4805259/c1059e506aeb/pone.0151665.g001.jpg

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