葡聚糖硫酸钠诱导溃疡性结肠炎对猪肝损伤的影响

doi:10.16431/j.cnki.1671-7236.2025.11.037

摘要/Abstract

摘要： 【目的】通过葡聚糖硫酸钠(dextran sulfate sodium，DSS)灌胃快速诱导巴马猪肠炎，探究肠肝轴失调对肝脏功能和基因表达的影响，为畜牧业中肠道-肝脏相关疾病的防控提供理论依据，同时为将该模型转化应用于人类肝炎医学模型提供科学依据。【方法】将16头120日龄巴马公猪随机分为对照组(Control)和DSS组，每组8头。DSS组公猪连续6 d 灌胃DSS溶液(1.25 g/kg BW)，对照组公猪灌胃等体积纯净水。试验第7天屠宰后采集血清和肝脏样本。通过HE、免疫荧光及Masson染色观察肝脏组织病理变化，检测血清中谷丙转氨酶(ALT)、谷草转氨酶(AST)活性。对肝脏样本进行转录组测序，筛选DSS诱导肠炎后肝脏组织中差异表达基因(differentially expressed genes，DEGs)，并对其进行GO功能、KEGG通路以及GSEA富集分析，利用实时荧光定量PCR检测肝脏促炎及促纤维化相关基因表达量。【结果】HE染色显示，DSS组猪肝脏组织出现免疫细胞浸润增多、肝脏组织结构紊乱的现象。与对照组相比，DSS组猪血清中AST活性显著增加(P＜0.05)，ALT活性无显著变化(P＞0.05)。转录组分析筛选出2 456个DEGs(1 153个上调、1 303个下调)。GO功能注释显示，DEGs显著富集于炎症反应、脂肪酸代谢及胆固醇稳态等生物过程；KEGG通路富集分析显示，DEGs主要显著富集于胆汁酸代谢紊乱、PPAR信号通路抑制及NF-κB信号通路激活等。GSEA分析显示，上调通路多与炎症反应(如IL-6/JAK/STAT3、TNF-α/NF-κB等信号通路)相关，下调通路主要涉及代谢功能(如脂肪酸代谢、胆汁酸代谢等)。免疫荧光染色和实时荧光定量PCR结果显示，与对照组相比，DSS组猪肝脏CD45⁺细胞浸润和胶原沉积明显增多，促炎基因TNF-α、IL-6、CXCL2、CXCL10、MCP1和促纤维化基因COL3A1、TGF-β表达量显著升高(P＜0.05)。【结论】DSS灌胃能够快速诱导猪肝脏发生炎症，导致肝脏代谢功能紊乱，营养吸收受阻。本研究结果为畜禽肝脏疾病基础研究及人类肝炎医学模型转化提供了理论依据。

关键词: 猪; 肝脏; 葡聚糖硫酸钠(DSS); 转录组测序; 炎症; 纤维化

Abstract: 【Objective】 The purpose of this study was to rapidly induce enteritis in Bama pigs via dextran sulfate sodium (DSS) gavage and investigate the effects of gut-liver axis dysfunction on liver function and gene expression,aiming to provide a theoretical basis for the prevention and control of gut-liver related diseases in animal husbandry，and at the same time,provide scientific evidence for the transformation and application of this model for human hepatitis. 【Method】 Sixteen 120-day-old Bama boars were randomly divided into Control and DSS groups,with 8 boars in each group.The boars in DSS group were intragastrically administered with DSS solution (1.25 g/kg BW) for 6 consecutive days,while the boars in Control group were given the same volume of pure water by intragastric administration.On the 7th day of the experiment,serum and liver samples were collected after the animals were slaughtered.The pathological changes of liver tissues were observed through HE staining,immunofluorescence staining and Masson staining,and the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were detected.Transcriptome sequencing was performed on liver samples to screen for differentially expressed genes (DEGs) induced by DSS and to identify the genes that were differentially expressed in liver.GO function,KEGG pathway and GSEA enrichment analyses were performed.The expression of liver inflammatory and fibrotic-related genes were detected using Real-time quantitative PCR. 【Result】 HE staining showed that there was an increase in immune cell infiltration and disordered liver tissue structure in DSS group.Compared with Control group,the AST activity of serum in DSS group was significantly increased (P＜0.05),while the ALT activity showed no significant change (P＞0.05).Transcriptome analysis identified 2 456 DEGs (1 153 upregulated and 1 303 downregulated).GO function annotation showed that the DEGs were significantly enriched in biological processes such as inflammatory response,fatty acid metabolism,and cholesterol metabolism.KEGG pathway enrichment analysis revealed that the DEGs were mainly significantly enriched in disorders of bile acid metabolism,inhibition of the PPAR signaling pathway,and activation of the NF-κB signaling pathway,etc.GSEA analysis revealed that the upregulated pathways were mostly related to inflammatory responses (IL-6/JAK/STAT3 signaling pathway,TNF-α/NF-κB signaling axis,etc.),while the downregulated pathways mainly involved metabolic functions (fatty acid metabolism,bile acid metabolism,etc.).The results of immunofluorescence staining and Real-time quantitative PCR showed that compared with Control group,the infiltration of CD45^＋ cells and collagen deposition of liver in DSS group were significantly increased,and the expression of pro-inflammatory genes TNF-α,IL-6, CXCL2,CXCL10 and MCP1，and pro-fibrotic genes COL3A1 and TGF-β were significantly increased (P＜0.05). 【Conclusion】 DSS gavage could rapidly induce inflammation in porcine liver,leading to liver metabolic dysfunction and impaired nutrient absorption.The results provided a theoretical basis for basic research on liver diseases in livestock and poultry and for the translation of medical models of human hepatitis.

Key words: pig; liver; dextran sulfate sodium (DSS); RNA-Seq; inflammation; fibrosis

中图分类号:

S858.28

吕启榀, 彭渝, 韩煦, 杨述林, 王彦芳, 陶聪, 曹果清. 葡聚糖硫酸钠诱导溃疡性结肠炎对猪肝损伤的影响[J]. 中国畜牧兽医, 2025, 52(11): 5422-5433.

LYU Qipin, PENG Yu, HAN Xu, YANG Shulin, WANG Yanfang, TAO Cong, CAO Guoqing. Effects of DSS-induced Ulcerative Colitis on Liver Injury in Pigs[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5422-5433.

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