动物氨基酸转运载体的研究进展

氨基酸转运载体(AAT)是一类介导氨基酸从细胞外转运到细胞内的重要蛋白,也是一类能介导氨基酸相关的信号通路的重要营养物质感受分子,在机体的生长代谢、营养健康等方面具有重要作用。动物机体中存在多种类型的AAT,它们能感知机体内相关氨基酸水平的变化,介导细胞氨基酸感知信号通路——哺乳动物雷帕霉素靶蛋白复合体1(mTORC1)和一般性调控阻遏蛋白激酶2(GCN2)的激活,从而引起通路下游发挥作用。在不同组织细胞中,发挥主导作用的AAT存在差异,表明AAT具有组织特异性,同时,AAT也受多种因素的影响,比如动物机体本身、营养物质水平、激素水平等。作者主要从AAT的类型及转运机制、介导营养信号启动及对mTORC1通路和GCN通路的影响、在不同组织中的作用及AAT表达的调控4个方面进行综述,从宏观方面介绍了AAT,旨在为AAT的研究提供一些参考。     

动物胃肠道氨基酸感应与转运

动物胃肠道中不同类型的内分泌细胞构成了"胃肠道内分泌系统"。饲粮中的蛋白质在动物胃肠道被分解为氨基酸后,能够被内分泌细胞膜上的氨基酸感应受体所识别,介导激素的分泌,调控胃肠道生理活动。位于肠上皮细胞的氨基酸转运载体通过哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)和一般性调控阻遏蛋白激酶2(GCN2)信号通路调控细胞氨基酸代谢。本文就动物胃肠道内分泌系统、氨基酸感应受体与转运载体以及氨基酸感知信号通路等方面研究进展进行综述。     

动物氨基酸转运与感知系统研究进展

氨基酸转运载体既有转运活性,又可作为感受器发挥胞外氨基酸感知功能。细胞膜上的氨基酸转运载体,尤其是转运大中性氨基酸包括亮氨酸的转运载体,能够通过胞内营养信号通路,包括调控细胞生长的哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)通路以及被氨基酸饥饿所激活的一般性调控阻遏蛋白激酶(GCN)通路,调控细胞代谢。鉴于氨基酸转运载体的研究对动物营养学的重要性,本文对氨基酸转运载体的分类、氨基酸转运载体介导的氨基酸感知功能及氨基酸转运载体的组织特异性进行综述,以期更好的协助相关研究的发展。     

乳腺对氨基酸的摄取及其调控机制研究进展

乳蛋白是乳中重要的营养成分之一,超过90%的乳蛋白是乳腺利用从血液中摄取的氨基酸从头合成,因此在保证氨基酸充足供给的前提下,乳腺对氨基酸摄取率的高低是影响乳蛋白产量的关键因素。血液中的氨基酸不能自由扩散进出乳腺,需要由乳腺上皮细胞膜上特异的氨基酸转运载体(AAT)协助完成。而乳腺AAT活性受到营养物质和激素水平的调节,当乳腺感知到营养物质和激素水平变化的信号,能够通过激活或抑制以哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)和一般性调控阻遏蛋白激酶2(GCN2)为核心的2条信号通路的活性,进而影响AAT活性,调节乳腺对氨基酸的摄取。本文主要从乳腺AAT的分类和功能、影响乳腺摄取氨基酸的主要因素以及调控乳腺氨基酸摄取的信号通路机制3个方面作一综述,旨在从氨基酸摄取的角度为提高乳蛋白的合成提供参考。     

GCN2和mTORC1信号路径对动物机体氨基酸平衡的调节作用

氨基酸平衡对哺乳动物健康生长具有重要意义,而在维持机体氨基酸平衡的过程中,GCN2和mTORC1信号路径发挥着重要作用。GCN2路径能有效感应胞内氨基酸缺乏,而mTORC1路径则能对胞外氨基酸水平的变化做出响应。本文结合近年来有关GCN2和mTORC1的研究进展,阐述了氨基酸充足和氨基酸饥饿条件下,GCN2和mTORC1这两条信号路径的感应特点以及随后启动的相关应答机制,包括蛋白质合成、拒食行为、氨基酸转运体表达增加、氨基酸合成酶增加和自噬启动等。了解动物在不同的氨基酸营养状态下维持细胞内氨基酸平衡的这些调节机制,有助于人们更好的对动物进行氮营养调控,从而改善肠道健康。     

氨基酸调节哺乳动物雷帕霉素靶蛋白复合体1信号通路的分子机制

哺乳动物雷帕霉素靶蛋白复合体1(mTORC1)信号通路能够感受一系列细胞内外环境因素的变化,如氨基酸浓度、能量水平、生长因子等进而调节细胞生长。氨基酸不仅是合成蛋白质的底物,也可作为信号分子激活mTORC1信号通路,促进蛋白质合成。溶酶体是氨基酸激活mTORC1信号通路过程中一个重要细胞器,mTORC1感应氨基酸的上游信号通路需要溶酶体相关蛋白及胞浆蛋白的参与完成。本文综述了氨基酸调节mTORC1信号通路的分子机制,为营养因子调控蛋白质合成的关键通路提供参考。     

葡萄糖感应机制与肠道内分泌调控的研究进展

动物肠道具有感应肠腔葡萄糖的功能,机体通过葡萄糖激酶(GCK)、味觉受体、葡萄糖转运蛋白GLUT2、mTORC1信号通路及AMPK等机制感应葡萄糖,影响肠道内分泌细胞(enteroendocrine cells,EECs)分泌激素,形成复杂的内分泌调控网络,调节机体营养物质代谢和采食行为等重要生理活动。文章综述了动物葡萄糖的感应机制及其对肠道内分泌调控的影响。     

谷氨酸吸收转运及对肠道发育影响的研究进展

谷氨酸作为肠上皮主要的能源物质,为肠黏膜生理功能的正常实现(包括营养物质的吸收转运和信号传导的进行以及黏膜上皮细胞的自身更新和高度有序结构的维持)供给能量。同时,其作为一种重要的信号分子和功能性氨基酸,可激活哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)信号通路,参与蛋白质合成,促进细胞增殖,增强肠道抗氧化能力,进而保护肠上皮结构和功能的完整性,促进肠道发育。本文就谷氨酸的吸收转运和代谢系统及其对mTORC1信号通路和动物肠道黏膜屏障功能的影响作一综述,旨在为谷氨酸功能的挖掘及调控动物肠道发育的生产应用提供参考。     

谷氨酸吸收转运及对肠道发育影响的研究进展北大核心CSCD

谷氨酸作为肠上皮主要的能源物质,为肠黏膜生理功能的正常实现(包括营养物质的吸收转运和信号传导的进行以及黏膜上皮细胞的自身更新和高度有序结构的维持)供给能量。同时,其作为一种重要的信号分子和功能性氨基酸,可激活哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)信号通路,参与蛋白质合成,促进细胞增殖,增强肠道抗氧化能力,进而保护肠上皮结构和功能的完整性,促进肠道发育。本文就谷氨酸的吸收转运和代谢系统及其对mTORC1信号通路和动物肠道黏膜屏障功能的影响作一综述,旨在为谷氨酸功能的挖掘及调控动物肠道发育的生产应用提供参考。     

小肽转运载体的生物学特征、影响因素及对肠道稳态的调控

小肽转运载体介导的小肽的吸收在促进动物的生长发育和提高动物生产性能中发挥着重要作用。肠道作为动物营养物质消化吸收的主要部位，肠道内环境的稳态对动物机体的健康和生长发育至关重要。由于小肽转运载体参与营养物质转运及调控肠道稳态与肠道炎症，所以肽转运蛋白成为了营养学、生理学、药理学上的研究焦点。本文就小肽转运载体的结构、转运机制、功能、表达及活性调控进行了综述，特别总结了小肽转运载体1在肠道炎症与调控肠道稳态中的作用。     

氨基酸缺乏诱导细胞自噬的哺乳动物雷帕霉素靶蛋白复合体C1信号通路机制研究进展

自噬是机体维持自身稳态的一种重要生理活动,当体内氨基酸或葡萄糖等营养缺乏时,细胞会启动自噬。自噬受到多种信号通路的调节,哺乳动物雷帕霉素靶蛋白复合体C1(m TORC1)信号通路是其中重要的一条,它可以使自噬相关基因13(Atg13)磷酸化,抑制自噬起始。本文将围绕近年来报道的氨基酸缺乏诱导细胞自噬的m TORC1信号通路,包括小G蛋白、腺苷酸活化蛋白激酶(AMPK)、微小RNA(miRNA)、氨基酰-tRNA合成酶在其中的作用等研究进展进行综述。     

Regulation of mTORC1 by amino acids in mammalian cells: A general picture of recent advances

The mechanistic target of rapamycin complex 1 (mTORC1) integrates various types of signal inputs, such as energy, growth factors, and amino acids to regulate cell growth and proliferation mainly through the 2 direct downstream targets, eukaryotic translation initiation factor 4E-binding protein 1 (4EBP1) and ribosomal protein S6 kinase 1 (S6K1). Most of the signal arms upstream of mTORC1 including energy status, stress signals, and growth factors converge on the tuberous sclerosis complex (TSC) − Ras homologue enriched in brain (Rheb) axis. Amino acids, however, are distinct from other signals and modulate mTORC1 using a unique pathway. In recent years, the transmission mechanism of amino acid signals upstream of mTORC1 has been gradually elucidated, and some sensors or signal transmission pathways for individual amino acids have also been discovered. With the help of these findings, we propose a general picture of recent advances, which demonstrates that various amino acids from lysosomes, cytoplasm, and Golgi are sensed by their respective sensors. These signals converge on mTORC1 and form a huge and complicated signal network with multiple synergies, antagonisms, and feedback mechanisms.     

亮氨酸对猪胎盘滋养层细胞增殖及氨基酸转运的影响

为研究亮氨酸(Leu)对猪胎盘滋养层细胞(pTr)增殖、凋亡以及氨基酸转运载体表达的影响及其机制,本试验用不同浓度Leu(0、1、10 mmol/L)分别处理pTr细胞24 h和48 h后,使用荧光定量PCR技术检测pTr细胞增殖和凋亡相关基因、氨基酸转运载体以及mTOR信号通路关键蛋白等的mRNA表达水平。结果表明:Leu处理pTr细胞24 h后,1 mmol/L试验组的SNAT1(P<0.01)、4E-BP1 (P<0.05)和eIF4G(P<0.05)的mRNA相对表达量低于对照组;Leu处理pTr细胞48 h后,1 mmol/L试验组LAT1(P<0.05)、4E-BP1(P<0.01)的mRNA相对表达量低于对照组,10 mmol/L试验组CDK4(P<0.05)、4E-BP1 (P <0.01)、SNAT1 (P <0.01)、SNAT2 (P <0.01)、LAT1 (P <0.01)以及rBAT (P <0.05)的mRNA相对表达量也低于对照组;Leu处理pTr细胞24 h和48 h后,10 mmol/L组mTORC1的mRNA相对表达量较对照组和1 mmol/L组均极显著提高(P<0.01)。可见,10 mmol/L Leu会抑制pTr细胞的增殖活力,并可能通过mTOR信号通路的介导,降低了pTr细胞氨基酸转运载体的表达。     

Postnatal development of nutrient transport in the intestine of dogs

OBJECTIVE: To measure nutrient absorption by the intestine during postnatal development of dogs. ANIMAL: 110 Beagles ranging from neonatal to adult dogs. PROCEDURE: Rates of absorption for sugars (glucose, galactose, and fructose), amino acids (aspartate, leucine, lysine, methionine, and proline), a dipeptide (glycyl-sarcosine), and linoleic acid by the proximal, mid, and distal regions of the small intestine were measured as functions of age and concentration (kinetics) by use of intact tissues and brush-border membrane vesicles. Absorption of octanoic acid by the proximal portion of the colon was measured in intact tissues. RESULTS: Rates of carrier-mediated transport by intact tissues decreased from birth to adulthood for aldohexoses and most amino acids but not for fructose and aspartate. Kinetics and characteristics of absorption suggest that there were changes in the densities, types, and proportions of various carriers for sugars and amino acids. Saturable absorption of linoleic acid in the small intestine and octanoic acid in the proximal portion of the colon increased after weaning. CONCLUSIONS AND CLINICAL RELEVANCE: Rates of absorption decreased between birth and adulthood for most nutrients. However, because of intestinal growth, absorption capacities of the entire small intestine remained constant for leucine and proline and increased for glucose, galactose, fructose, aspartate, and proline but were less than predicted from the increase in body weight. Although postnatal ontogeny of nutrient absorption was consistent with changes in the composition of the natural and commercial diets of growing dogs, rates of amino acid and peptide absorption were lower than expected.     

日粮组成对牛奶中乳成分合成与调控机制影响的研究

牛奶中乳蛋白和乳脂肪含量的高低直接关系到牛奶的品质和风味。牛奶中乳蛋白主要可以分为酪蛋白和乳清蛋白两种类型,其合成代谢过程均受到mTOR信号分子通路、JAK-STAT信号分子通路、GCN2-eIF2a信号分子通路的影响。牛奶中乳脂肪主要为三酰基甘油酯、磷脂等,对于牛奶的营养和风味均有重要影响,其受到脂肪酸相关酶ACACA、FAS、SCD1的调控。饲料营养是影响牛奶中乳蛋白和乳脂肪的关键因素之一,包括精粗饲料配比及饲料添加剂的应用等方面。本文对牛奶中乳蛋白和乳脂肪的合成调控机理,及日粮组成对其的影响机制进行阐述,为改善乳品质提供参考。     

系统分析多组织转录组鉴定影响猪脂肪沉积的关键基因

旨在挖掘影响松辽黑猪脂肪沉积的关键基因及脂肪、肝和肌肉在体内的功能。本研究选择体重100 kg左右健康且背膘厚差异显著的6头(高、低各3头)松辽黑猪为试验动物,利用高通量转录组测序技术检测其脂肪、肝和背最长肌组织中基因的表达水平,鉴定不同脂肪沉积猪和不同组织中的差异表达基因,并分析差异表达基因的生物学功能。结果表明,在不同分组的猪中发现135个差异表达基因,其中部分参与了PPAR信号通路、AMPK信号通路、代谢通路、脂肪酸代谢和甘油代谢等通路。经生物学功能分析发现,EHHADH、ME1、SCD、OLR1、PHGDH、ACLY、LEP和CYP超家族基因等基因为影响猪脂肪沉积的关键基因。在不同组织的差异表达表达基因中,脂肪组织中高表达的基因显著富集在胰岛素信号通路、MAPK信号通路、三羧酸循环、氧化磷酸化等通路;肝中高表达基因显著富集在多种物质的代谢、脂肪酸的降解、氨基酸的合成等通路;背最长肌中高表达的基因主要参与了蛋白质的降解、PI3K-Akt信号通路、氧化磷酸化通路、Wnt信号通路、磷脂酰肌醇信号通路等通路。不同组间差异表达基因分析结果提示,EHHADH、ME1、SCD、OLR1、PHGDH、ACLY、LEP和CYP超家族基因等基因是影响脂肪沉积的关键基因;不同组织间差异表达基因表明,脂肪组织是脂肪合成的主要部位,而肝和肌肉组织主要涉及脂肪酸的降解。本研究结果对脂肪性状的遗传改良、机理解析有一定意义。     

The market for amino acids: understanding supply and demand of substrate for more efficient milk protein synthesis

For dairy production systems, nitrogen is an expensive nutrient and potentially harmful waste product. With three quarters of fed nitrogen ending up in the manure, significant research efforts have focused on understanding and mitigating lactating dairy cows' nitrogen losses. Recent changes proposed to the Nutrient Requirement System for Dairy Cattle in the US include variable efficiencies of absorbed essential AA for milk protein production. This first separation from a purely substrate-based system, standing on the old limiting AA theory, recognizes the ability of the cow to alter the metabolism of AA. In this review we summarize a compelling amount of evidence suggesting that AA requirements for milk protein synthesis are based on a demand-driven system. Milk protein synthesis is governed at mammary level by a set of transduction pathways, including the mechanistic target of rapamycin complex 1(mTORC1), the integrated stress response(ISR), and the unfolded protein response(UPR). In tight coordination, these pathways not only control the rate of milk protein synthesis, setting the demand for AA, but also manipulate cellular AA transport and even blood flow to the mammary glands, securing the supply of those needed nutrients. These transduction pathways, specifically mTORC1, sense specific AA, as well as other physiological signals, including insulin, the canonical indicator of energy status. Insulin plays a key role on mTORC1 signaling, controlling its activation, once AA have determined mTORC1 localization to the lysosomal membrane.Based on this molecular model, AA and insulin signals need to be tightly coordinated to maximize milk protein synthesis rate. The evidence in lactating dairy cows supports this model, in which insulin and glucogenic energy potentiate the effect of AA on milk protein synthesis. Incorporating the effect of specific signaling AA and the differential role of energy sources on utilization of absorbed AA for milk protein synthesis seems like the evident following step in nutrient requirement systems to further improve N efficiency in lactating dairy cow rations.     

Amino acids regulate energy utilization through mammalian target of rapamycin complex 1 and adenosine monophosphate activated protein kinase pathway in porcine enterocytes

As major fuels for the small intestinal mucosa, dietary amino acids (AA) are catabolized in the mitochondria and serve as sources of energy production. The present study was conducted to investigate AA metabolism that supply cell energy and the underlying signaling pathways in porcine enterocytes. Intestinal porcine epithelial cells (IPEC-J2) were treated with different concentrations of AA, inhibitor, or agonist of mammalian target of rapamycin complex 1 (mTORC1) and adenosine monophosphate activated protein kinase (AMPK), and mitochondrial respiration was monitored. The results showed that AA treatments resulted in enhanced mitochondrial respiration, increased intracellular content of pyruvic acid and lactic acid, and increased hormone-sensitive lipase mRNA expression. Meanwhile, decreased citrate synthase, isocitrate dehydrogenase alpha, and carnitine palmitoyltransferase 1 mRNA expression were also observed. We found that AA treatments increased the protein levels of phosphorylated mammalian target of rapamycin (p-mTOR), phosphorylated-p70 ribosomal protein S6 kinase, and phosphorylated-4E-binding protein 1. What is more, the protein levels of phosphorylated AMPK α (p-AMPKα) and nicotinamide adenine dinucleotide (NAD)-dependent protein deacetylase sirtuin-1 (SIRT1) were decreased by AA treatments in a time depending manner. Mitochondrial bioenergetics and the production of tricarboxylic acid cycle intermediates were decreased upon inhibition of mTORC1 or AMPK. Moreover, AMPK activation could up-regulate the mRNA expressions of inhibitor of nuclear factor kappa-B kinase subunit beta (Ikbkβ), integrin-linked protein kinase (ILK), unconventional myosin-Ic (Myo1c), ribosomal protein S6 kinase beta-2 (RPS6Kβ2), and vascular endothelial growth factor (VEGF)-β, which are downstream effectors of mammalian target of rapamycin (mTOR). The mRNA expressions of phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit delta isoform (PIK3CD) and 5′-AMP-activated protein kinase subunit gamma-1 (PRKAG1), which are upstream regulators of mTOR, were also up-regulated by AMPK activation. On the other hand, AMPK activation also down-regulated FK506-binding protein 1A (FKBP1A), serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B beta isoform, phosphatase and tensin homolog (PTEN), and unc-51 like autophagy activating kinase 1 (Ulk1), which are up-stream regulators of mTORC1. Taken together, these data indicated that AA regulated cellular energy metabolism through mTOR and AMPK pathway in porcine enterocytes. These results demonstrated interactions of AMPK and mTORC1 pathways in AA catabolism and energy metabolism in intestinal mucosa cells of piglets, and also provided reference for using AA to remedy human intestinal diseases.     

Novel metabolic and physiological functions of branched chain amino acids: a review

It is widely known that branched chain amino acids(BCAA) are not only elementary components for building muscle tissue but also participate in increasing protein synthesis in animals and humans. BCAA(isoleucine, leucine and valine) regulate many key signaling pathways, the most classic of which is the activation of the m TOR signaling pathway. This signaling pathway connects many diverse physiological and metabolic roles. Recent years have witnessed many striking developments in determining the novel functions of BCAA including:(1) Insufficient or excessive levels of BCAA in the diet enhances lipolysis.(2) BCAA, especially isoleucine, play a major role in enhancing glucose consumption and utilization by up-regulating intestinal and muscular glucose transporters.(3)Supplementation of leucine in the diet enhances meat quality in finishing pigs.(4) BCAA are beneficial for mammary health, milk quality and embryo growth.(5) BCAA enhance intestinal development, intestinal amino acid transportation and mucin production.(6) BCAA participate in up-regulating innate and adaptive immune responses.In addition, abnormally elevated BCAA levels in the blood(decreased BCAA catabolism) are a good biomarker for the early detection of obesity, diabetes and other metabolic diseases. This review will provide some insights into these novel metabolic and physiological functions of BCAA.