第72期：丁酸对饲喂以玉米或小麦为基础日粮的肉鸡胰岛素稳态的影响

肠动力研究院Service

Effects of butyrate on the insulin homeostasis of chickens kept on maize-or wheat-based diets

作者：Anna KULCSÁR1^*, Gábor MÁTIS¹ , Andor MOLNÁR², Janka PETRILLA¹ , Ferenc HUSVÉTH² , Korinna HUBER^3a, Károly DUBLECZ² and Zsuzsanna NEOGRÁDY¹
¹Department of Physiology and Biochemistry, University of Veterinary Medicine,
István u. 2, H-1078 Budapest, Hungary;
²Department of Animal Sciences and Animal Husbandry, Georgikon Faculty, University of Pannonia, Keszthely, Hungary;
³Department of Physiology, University of Veterinary Medicine, Hanover, Germany;
^aPresent address: Institute of Animal Science, University of Hohenheim,
Stuttgart-Hohenheim, Germany

来源：Acta Veterinaria Hungarica 64 (4), pp. 482-496 (2016)
DOI: 10.1556/004.2016.045

翻译：肠动力研究院梁琦

【摘要】丁酸已被证明可通过供给胃肠道上皮细胞能量，调节细胞的增殖和分化，增强肠道的吸收能力和维持肠道菌群稳态等途径来改善动物肠道健康。然而，当前尚未有人阐明日粮类型（以玉米或小麦为基础饲粮）和/或饲料添加剂丁酸通过影响胰岛素信号传导来调控鸡内分泌代谢的分子机制，本实验旨在研究饲料添加剂丁酸对饲喂以玉米或小麦为基础饲粮的Ross 308肉鸡胰岛素信号蛋白（代谢和生长的强效调节剂）表达的影响。176羽1日龄的Ross 308肉用仔鸡被随机分成8个处理组（22羽/组），试验准备了两种不同的基础饲粮，分别是含有玉米（MB）和小麦（WB）的基础日粮。以MB日粮为主的处理组如下，1）MBC组：喂食玉米饲粮；2）MBB1.5组：饲喂玉米饲粮+1.5g/kg未保护的丁酸；3）MBB3组：喂食玉米饲粮+3g/kg未保护的丁酸；4）MBPB组：饲喂玉米饲粮+0.2g/kg微囊化包被的丁酸。以WB日粮为主的处理组如下，1）WBC组：喂食小麦饲粮；2）WBB1.5组：饲喂小麦日粮+1.5g/kg未保护的丁酸；3）WBB3组：喂食小麦日粮+3g/kg未保护的丁酸；4）WBPB组：饲喂小麦日粮+0.2g/kg微囊化包被的丁酸，试验为期42天。试验第42天，采取部分肉鸡的血液样品用于分析葡萄糖和胰岛素浓度，并取组织样品（肝脏，腓肠肌和皮下脂肪组织）进行蛋白质印迹（WB）检查。通过对所述组织的半定量WB来评估关键胰岛素信号传导蛋白（IRβ，PKCζ和mTOR）的表达。结果显示，日粮类型对鸡的胰岛素稳态有显著影响。与饲喂玉米日粮的动物相比，喂食小麦日粮可显著增加肝脏中IRβ和mTOR的表达以及脂肪组织中的mTOR和PKCζ的表达。除此外，还发现较低剂量（1.5g/kg）的未受保护的丁酸可刺激肝脏中IRβ的表达，这表明饲料添加剂丁酸可影响胰岛素的敏感性。研究结果表明，通过比较饲料添加剂丁酸的特殊效果和日粮类型，显示出丁酸作为营养因素的新的应用前景，试验猜测日粮中非淀粉性多糖（NSP）主要通过促进肠道中短链脂肪酸（SCFAs，主要是丁酸）的释放来影响鸡的胰岛素稳态。与哺乳动物相比，鸡的组织在生理上具有较低的胰岛素敏感性，因此，通过日粮调控胰岛素信号传导途径来改善肉鸡的生长和代谢健康可能特别重要。
【关键词】丁酸；胰岛素信号；肉用仔鸡

以下是试验相关图表

表1：在研究的各个阶段的试验日粮成分和计算的营养成分

表2：不同日粮喂养的肉鸡的平均体重和平均日采食量

图1：日粮类型和添加一定量的丁酸对肉用仔鸡血浆中葡萄糖和胰岛素浓度的影响。A：葡萄糖；B：胰岛素。MBC组：喂食玉米饲粮；MBB1.5组：饲喂玉米饲粮+1.5g/kg未保护的丁酸；MBB3组：喂食玉米饲粮+3g/kg未保护的丁酸；MBPB组：饲喂玉米饲粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。WBC组：喂食小麦饲粮；WBB1.5组：饲喂小麦日粮+1.5g/kg未保护的丁酸；WBB3组：喂食小麦日粮+3g/kg未保护的丁酸；WBPB组：饲喂小麦日粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。列中的星号表示通过试验测试获得的显著差异，括号中的星号表示日粮的显著主要影响。^*** P <0.001；^** P <0.01；^*P <0.05；^#P <0.10

图2.1：日粮类型和添加一定量的丁酸对肉用仔鸡肝脏中胰岛素信号蛋白的影响。纵轴（y）表示（A）IRβ，（B）mTOR和（C）PKCζ蛋白表达的标准化痕量值。MBC组：喂食玉米饲粮；MBB1.5组：饲喂玉米饲粮+1.5g/kg未保护的丁酸；MBB3组：喂食玉米饲粮+3g/kg未保护的丁酸；MBPB组：饲喂玉米饲粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。WBC组：喂食小麦饲粮；WBB1.5组：饲喂小麦日粮+1.5g/kg未保护的丁酸；WBB3组：喂食小麦日粮+3g/kg未保护的丁酸；WBPB组：饲喂小麦日粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。列中的星号表示通过试验测试获得的显著差异，括号中的星号表示日粮的显著主要影响。 ^*** P <0.001；^** P <0.01；^*P <0.05；^#P <0.10。

图2.2：日粮类型和添加一定量的丁酸对肉用仔鸡脂肪组织中胰岛素信号蛋白的影响。纵轴（y）表示（A）IRβ，（B）mTOR和（C）PKCζ蛋白表达的标准化痕量值。MBC组：喂食玉米饲粮；MBB1.5组：饲喂玉米饲粮+1.5g/kg未保护的丁酸；MBB3组：喂食玉米饲粮+3g/kg未保护的丁酸；MBPB组：饲喂玉米饲粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。WBC组：喂食小麦饲粮；WBB1.5组：饲喂小麦日粮+1.5g/kg未保护的丁酸；WBB3组：喂食小麦日粮+3g/kg未保护的丁酸；WBPB组：饲喂小麦日粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。列中的星号表示通过试验测试获得的显著差异，括号中的星号表示日粮的显著主要影响。^***P <0.001；^** P <0.01；^*P <0.05；^#P <0.10。

图2.3:日粮类型和添加一定量的丁酸对肉用仔鸡骨骼肌胰岛素信号蛋白的影响。纵轴（y）表示（A）IRβ，（B）mTOR和（C）PKCζ蛋白表达的标准化痕量值。MBC组：喂食玉米饲粮；MBB1.5组：饲喂玉米饲粮+1.5g/kg未保护的丁酸；MBB3组：喂食玉米饲粮+3g/kg未保护的丁酸；MBPB组：饲喂玉米饲粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。WBC组：喂食小麦饲粮；WBB1.5组：饲喂小麦日粮+1.5g/kg未保护的丁酸；WBB3组：喂食小麦日粮+3g/kg未保护的丁酸；WBPB组：饲喂小麦日粮+0.2g/kg微囊化包被的丁酸（Butipearl®）。列中的星号表示通过试验测试获得的显著差异，括号中的星号表示日粮的显著主要影响。^***P <0.001；^** P <0.01；^*P <0.05；^#P <0.10。

结论

总之，本研究描述了通过营养影响鸡胰岛素稳态的新方法，例如作为饲料添加剂的丁酸，以及应用各种谷物作为具有不同NSP水平的膳食碳水化合物来源。应该强调的是，使用基于小麦的饲粮（具有较高的可溶性NSP水平）可通过刺激包括丁酸在内的SCFA在肠道内的生成，这可能对胰岛素稳态具有强烈影响。基于这些发现，通过比较不同的应用形式和描述一些潜在的分子机制不难得出SCFA（主要是丁酸）是一种内分泌代谢的有效调节物。研究发现，添加较高的膳食可溶性NSP水平和/或饲料添加剂丁酸可能是家禽养殖中一种有效工具，它们主要通过影响胰岛素稳态，从而改善新陈代谢，生长发育和动物健康。

Abstract

The aim of the present study was to investigate the effects of butyrate as a feed supplement on the expression of insulin signalling proteins as potent regulators of metabolism and growth in Ross 308 broiler chickens fed maize- or wheat-based diets. Both diets were supplemented with non-protected butyrate (1.5 and 3.0 g/kg of diet, respectively) or with protected butyrate (0.2 g/kg of diet); the diet of the control groups was prepared without any additives (control). On day 42 of life, systemic blood samples were drawn for analyses of glucose and insulin concentrations, and tissue samples (liver, gastrocnemius muscle and subcutaneous adipose tissue) were taken for Western blotting examinations. The expression of key insulin signalling proteins (IRβ, PKCζ and mTOR) was assessed by semiquantitative Western blotting from the tissues mentioned. The type of diet had a remarkable influence on the insulin homeostasis of chickens. The wheat-based diet significantly increased IRβ and mTOR expression in the liver as well as mTOR and PKCζ expression in the adipose tissue when compared to animals kept on a maize-based diet. IRβ expression in the liver was stimulated by the lower dose of non-protected butyrate as well, suggesting the potential of butyrate as a feed additive to affect insulin sensitivity. Based on the results obtained, the present study shows new aspects of nutritional factors by comparing the special effects of butyrate as a feed additive and those of the cereal type, presumably in association with dietary non-starch polysaccharide- (NSP-) driven enteric short-chain fatty acid release including butyrate, influencing insulin homeostasis in chickens. As the tissues of chickens have physiologically lower insulin sensitivity compared to mammals, diet-associated induction of the insulin signalling pathway can be of special importance in improving growth and metabolic health.
Key words: Butyrate, insulin signalling, broiler chicken

Conclusion

Concluding our results, the present study describes new ways of influencing insulin homeostasis of chickens by nutrition, such as by butyrate as a feed additive and by applying various cereals as dietary carbohydrate sources with different NSP levels. It should be stressed that the application of a wheat-based diet (with higher soluble NSP levels) may have a strong influence on insulin homeostasis by stimulating the intestinal production of SCFA including butyrate. Based on these findings, the role of SCFA as potent effectors of endocrine metabolic regulation, and primarily that of butyrate, was highlighted by comparing different application forms and by describing some underlying molecular mechanisms. Concerning the data obtained, applying higher dietary soluble NSP levels and/or butyrate as a feed additive can be a promising tool in poultry farming to influence insulin homeostasis and thus improve metabolism, growth and animal health.

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