Interpretation of Fiber Supplementation on Offffspring Testicular Development in a Pregnant Sow Model from a Proteomics Perspective
作者：Yan Lin 1,2,*,†, Lujie Li 1,2,†, Yang Li 1,2,†, Ke Wang 1,2, Dongqin Wei 1,2, Shengyu Xu 1,2, Bin Feng 1,2 , Lianqiang Che 1,2, Zhengfeng Fang 1,2, Jian Li 1,2, Yong Zhuo 1,2 and De Wu 1,2,*
1 Key Laboratory of Animal Disease-Resistance Nutrition and Feed Science, Ministry of Agriculture, Sichuan Agricultural University, Chengdu 611130, China;
2 Key Laboratory of Animal Disease-Resistance Nutrition, Ministry of Education, Wenjiang 611130, China
来源：Int. J. Mol. Sci. 2019, 20, 4549; doi:10.3390/ijms20184549
翻译：肠动力研究院 梁琦
【摘要】膳食纤维是一种重要的营养素，研究发现膳食纤维在维持人和动物的健康、繁殖性能上扮演着一个重要的角色。但是，目前尚不清楚妊娠期间补充母体纤维营养对后代睾丸发育的影响，本文旨在研究妊娠母猪补充膳食纤维对雄性仔猪睾丸发育的影响及其可能机制。36头青年母猪（大白×约克夏，9月龄）被随机均分成2组，试验分组如下，1）CON组：妊娠期饲喂玉米-豆粕基础日粮；2）Fiber组：妊娠期喂食基础日粮+22.6g/kg菊粉+181.6g/kg纤维素；每组18头，试验为期118天。与此同时，母猪在妊娠期间限饲以防止体重超标。母猪分娩后，记录7日龄仔猪的体重和睾丸重量，同时收集睾丸用于进一步分析。结果显示，结果显示，Fiber组仔猪的睾丸器官指数和单个输精管中的精原细胞数量显著高于CON组（p <0.05）。另外，在Fiber组中还发现仔猪睾丸中的葡萄糖，乳酸和脂质的浓度显著增加（p <0.05）。TMT定量蛋白质组学分析表明，CON组和Fiber组的仔猪之间在糖脂转运，代谢，氧化和雄性生殖相关蛋白表达方面存在显著差异（p <0.05）。通过KEGG数据库鉴定出Fiber组仔猪的睾丸中存在最丰富的信号传导通路包括淀粉和蔗糖代谢，脂肪酸代谢，谷胱甘肽代谢和肾素-血管紧张素系统。实时定量PCR对mRNA表达分析进一步证实了某些信号通路在母体纤维营养调节后代睾丸发育中的重要性。总之，研究结果揭示了母体纤维营养对后代睾丸发育的潜在分子机制，并为今后探究母体纤维营养对人生殖性能的影响提供了富有意义的借鉴与参考。
【关键词】膳食纤维；母体；蛋白质组学；睾丸发育
以下是试验相关表格
图1：母体纤维营养对后代体重的影响（A），（B）后代睾丸器官指数。该值表示为平均值±SEM。与正常对照组相比，** p <0.01。Con =对照组，Fiber =纤维组。
图2: HE染色的仔猪睾丸组织横截面。（A）对照组小猪的睾丸，×400。（B）纤维组仔猪的睾丸，400×。（a）代表睾丸支持细胞，（b）代表精原细胞。
图3：母体纤维营养对后代睾丸中每个生精小管中精子和精细胞数量的影响。该值表示为平均值±SEM。与正常对照组相比，*表示P<0.05。Con =对照组，Fiber =纤维组。
图4：母体纤维营养对后代血液生化指标的影响。该值表示为平均值±SEM。与正常对照组相比，*表示P<0.05，**表示p <0.01。Con =对照组，Fiber =纤维组。GLU＝葡萄糖，CHO＝胆固醇，TG＝甘油三酯，HDL＝高密度脂蛋白，LDL＝低密度脂蛋白，L-LAC ＝L-乳酸酯，NEFA＝非酯化脂肪酸和GSH＝L-谷胱甘肽。
表1：纤维组和对照组的仔猪睾丸与KEGG信号通路有关的差异表达蛋白（DEP）。
图5：上调的DEP（A）和下调的DEP（B）的基因本体（GO）注释功能多样。直方图上的数字表示富集试验得到的p值的负对数变换（使用Fisher's精确检验）
图6：KEGG信号通路的DEPs富集分析。有15个DEP可以进行富集。其中，造血细胞谱系，肾素-血管紧张素系统以及淀粉和蔗糖代谢通路最丰富。
图7：母体纤维营养对相应mRNA的相对表达量的影响。该值表示为平均值±SEM。与正常对照组相比，*表示P<0.05，**表示p <0.01，***表示P<0.001。Con =对照组，Fiber =纤维组。
总结
通过相关试验，研究确定了影响睾丸细胞增殖的16种差异表达蛋白（DEP）。妊娠期间摄入不同膳食纤维的母猪的后代睾丸发育可能受到这些DEP所属通路的影响。对早期睾丸发育的影响可能是由于碳水化合物代谢，脂肪酸代谢和谷胱甘肽（GSH）代谢途径以及肾素-血管紧张素系统中关键点的调节。研究结果提高了我们对与母体纤维营养影响后代相关分子机制的理解。
Abstract
To study the effffects of maternal fifiber supplementation during pregnancy on the testicular development of male offffspring and its possible mechanisms, 36 sows (Landrace × Yorkshire) were allocated to either a control diet (n = 18) or a fifiber diet (the control diet supplemented with 22.60 g/kg inulin and 181.60 g/kg cellulosic; n = 18) during pregnancy. The body and testes weight of the offffspring, 7-day-old piglets, was recorded. Testes were collected for further analyses. Results showed that the testicular organ index and the number of spermatogonia in single seminiferous tubule were higher in piglets from the fifiber group than from the control group (p < 0.05). In addition, a signifificant increase in the concentration of glucose, lactate, and lipids in the testes was found in the fifiber group (p < 0.05). Proteomic analysis suggested that there were notable difffferences in glucolipid transport and metabolism, oxidation, and male reproduction-related proteins expression between the two groups (p < 0.05). Results revealed that the most enriched signaling pathways in the fifiber group testes included starch and sucrose metabolism, fatty acid metabolism, glutathione metabolism, and the renin-angiotensin system. mRNA expression analyzes further confifirmed the importance of some signaling pathways in maternal fifiber nutrition regulating offffspring testicular development. Our results shed new light on the underlying molecular mechanisms of maternal fifiber nutrition on offffspring testicular development and provided a valuable insight for future explorations of the effffect of maternal fifiber nutrition on man reproduction.
Keywords：fifiber；maternal；proteomics；testis development
Conclution
We identifified 16 difffferent DEPs which affffect testis cell proliferation. Testis development in the offffspring of sows with difffferent fifiber intake during gestation may be affffected by the pathways to which these DEPs belong. The exerted effffects on early testicle development may be due to the regulation of key points in carbohydrate metabolism, fatty acid metabolism, and GSH metabolism pathways as well as in the renin-angiotensin system. Our results improve our understanding of the molecular mechanisms associated to the effffect of maternal fifiber intake on offffspring.
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