| 易宏波 1，刘春艳 1,2，韦洋洋1,2，杨雪芬 1，黄艳娜 2，吴绮雯1，王 丽1，蒋宗勇1.猪源罗伊氏乳杆菌 LR1 对断奶仔猪肠黏膜细胞外基质动态变化的影响[J].广东农业科学,2023,50(11):17-28 |
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猪源罗伊氏乳杆菌 LR1 对断奶仔猪肠黏膜细胞外基质动态变化的影响 |
| Effect of Porcine Lactobacillus reuteri LR1 on Dynamic Changes of Extracellular Matrix in the Intestinal Mucosa of Weaned Piglets |
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| DOI：10.16768/j.issn.1004-874X.2023.11.002 |
| 中文关键词: 断奶仔猪 肠黏膜 炎症 肠道健康 细胞外基质 罗伊氏乳杆菌 LR1 |
| 英文关键词: weaned piglet intestinal mucosa inflammation intestinal health extracellular matrix Lactobacillus reuteri LR1 |
| 基金项目:国家自然科学基金（32272920）；广东省自然科学基金（2023A1515012491）；国家生猪产业技术体系专项（CAR-35）；广州市科技计划项目（202201011372）；岭南现代农业科学与技术广东省实验室茂名分中心基金项目（2022ZD003） |
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| 中文摘要: |
| 【目的】初步研究饲粮添加猪源罗伊氏乳杆菌对断奶仔猪肠道细胞外基质（Extracellular matrix，ECM）表达的影响，为猪源罗伊氏乳杆菌在畜牧业中的推广应用提供参考。【方法】选取 21 日龄体重相近的杜长大杂交断奶仔猪 144 头，随机分为 3 个处理组，对照组（CON）饲喂基础饲粮，抗生素组（AO）在基础饲粮中添加 75 mg/kg 金霉素和 100 mg/kg 喹乙醇，罗伊氏乳杆菌组（LR1）在基础饲粮中添加 5×1010 CFU/kg 猪源罗伊氏乳杆菌 LR1，每个处理组各分 8 栏饲养，每栏 6 头猪，试验期为 42 d，饲养试验结束时随机从每栏挑选 1 头仔猪进行屠宰，取其十二指肠（近端）、空肠（中段）、回肠（远端）样品，进行肠道胶原蛋白、纤维黏连蛋白、肌腱蛋白及其相关调控因子等 ECM 相关指标的检测。【结果】Masson 染色观察结果显示，十二指肠、空肠和回肠中胶原纤维均由肠黏膜下层向上层延伸，与 CON 组相比，LR1 组和 AO 组断奶仔猪十二指肠的胶原容积分数均显著降低（P<0.05），LR1 组的回肠胶原容积分数也显著降低（P<0.05）。TMT 高通量蛋白质组学分析结果显示，ECM-受体互作通路是差异蛋白主要富集通路之一，且其差异蛋白 COL1A2、COL4A2、COL6A1、ITGA1 等 ECM 分子的表达在 LR1 组显著低于 CON 组（P<0.05）。与 CON 组相比，LR1 组和 AO 组断奶仔猪空肠和回肠 COL1A2、COL3A1、ITGβ2 的基因表达均显著降低（P<0.05），且 LR1 组显著降低了空肠 COL4A2、COL5A1、COL6A1、FN1、TNC、ITGα1 及回肠 ITGα1 的基因表达（P<0.05），但对回肠 COL4A2、COL5A1、COL6A1、FN1、TNC 无显著影响。在回肠中，与 CON 组相比，LR1 组和 AO 组断奶仔猪回肠 Collagen Ⅰ ~ Ⅵ的含量显著降低（P<0.05）；LR1 组回肠胞内调控因子 SEC6A1、PDE4D 和胞外调控因子 MMP2 的基因表达均显著降低（P<0.05）；与 AO 组相比，LR1 组的胞内调控因子 SEC6A1、细胞因子 TGF-β 的基因表达显著降低（P<0.05）。【结论】饲粮中添加猪源罗
伊氏乳杆菌 LR1 可通过减少 ECM 调控因子 SEC6A1、PDE4D 等的基因表达来调节断奶仔猪肠道 ECM 的重塑。 |
| 英文摘要: |
| 【Objective】It aims to preliminarily reveal the effect of dietary supplementation with porcine Lactobacillus reuteri LR1 on the expression of intestinal extracellular matrix (ECM) in weaned piglets, providing references for the promotion and application of porcine LR1 in animal husbandry.【Method】One hundred and forty-four Duroc-Landrace-Yorkshire crossbred weaned piglets with similar body weights at 21 days of age were selected and randomly divided into three treatment groups, which were fed a basal diet (control group, CON group), a basal diet supplemented with 75 mg/kg of aureomycin and
100 mg/kg of quinol (antibiotic group, AO group), and a basal diet supplemented with 5×1010 CFU/kg of porcine L. reuteri LR1 (porcine L. reuteri LR1 group, LR1 group), respectively, and each treatment group was divided into 8 pens for feeding, with 6 pigs in each pen. The experiment lasted for 42 days. On the 43rd day, one piglet from each group was randomly selected for slaughter, and samples of the duodenum (proximal), jejunum (middle), and ileum (distal) were taken for the detection of extracellular matrix related indicators such as intestinal collagen, f ibronectin, tenascin and related regulatory factors.【Result】Masson staining observations showed that collagen fibers in the duodenum, jejunum and ileum all extended from the lower to the upper layers of the intestinal mucosa. Compared with the CON group, the collagen volume fraction of the duodenum decreased significantly in both the LR1 and AO groups (P<0.05), and the collagen volume fraction of the ileum also decreased significantly in the LR1 group (P<0.05). TMT high-throughput proteomics results showed that the ECM-receptor interaction pathway was one of the major differential protein-enriched pathways, and the expression of its differential proteins, COL1A2, COL4A2, COL6A1, ITGA1, and other ECM molecules, was significantly lower in the LR1 group than that in the CON group (P<0.05). In addition, the gene expression of COL1A2, COL3A1 and ITGβ2 in jejunum and ileum of LR1 and AO groups was significantly reduced compared with the CON group (P<0.05), and the gene expression of COL4A2, COL5A1, COL6A1, FN1, TNC, ITGα1 in jejunum, and ITGα1 in ileum was also significantly reduced in the LR1 group (P< 0.05), but there were no significant effect on COL4A2, COL5A1, COL6A1, FN1 and TNC in ileum. Compared with the CON group (P<0.05), in the ileum of the LR1 and AO groups, the contents of Collagen I, Collagen Ⅱ , Collagen Ⅲ , Collagen Ⅳ , Collagen Ⅴ and Collagen Ⅵ significantly decreased; the gene expression of ileal cellular regulatory factors SEC6A1 and PDE4D in LR1 group significantly decreased (P<0.05). Compared with the AO group (P<0.05), the gene expression of the extracellular regulator MMP2 was significantly lower in the LR1 group (P<0.05); and the gene expression of the intracellular regulator SEC6A1 and cytokine TGF-β significantly decreased in the LR1 group.【Conclusion】Addition of porcine L. reuteri LR1 to the diet could modulate ECM remodeling by reducing gene expression of ECM regulators SEC6A1, PDE4D and other genes. |
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