State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China; Department of Animal Genetics and Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
State Key Laboratory of Animal Biotech Breeding, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Int J Biol Macromol. 2024 Nov;281(Pt 3):136245. doi: 10.1016/j.ijbiomac.2024.136245. Epub 2024 Oct 3.
Intestinal disorders are common in metabolic syndrome. However, their pathogenesis is still not fully understood. Pig and human intestines are highly similar in terms of associated metabolic processes. Here, we successfully constructed a metabolic disease-susceptible transgenic (TG) Bama pig model by knocking in three humanized disease risk genes with the CRISPR/Cas9 technique to assess its potential as a model for human intestinal diseases and explore the possible pathological mechanisms involved. We found that jejunal barrier integrity was disrupted and that the infiltration of inflammatory cells increased in TG pigs after high-fat and high-sucrose diet (HFHSD) treatment. We revealed significant differences in the transcriptome, associated microbiome profiles and microbial metabolite short-chain fatty acid (SCFA) content of the jejunum of TG pigs. Notably, we found that SLC26A3 was significantly downregulated in TG pigs. Knockdown or overexpression of the SLC26A3 gene in IPEC-J2 cells significantly affected the expression of MUC2, MUC13 and occludin. Furthermore, in vitro experiments further verified that CDX2 directly regulated the expression of SLC26A3. Mechanistically, CDX2 mediated intestinal barrier function by enhancing the expression of SLC26A3 by binding to its promoter region between -1120 and - 1070 bp. TG pigs represent a promising model that provides new insights into preclinical research on human intestinal metabolic diseases associated with metabolic disorders and revealed that SLC26A3 may be a potential therapeutic target for intestinal metabolic diseases.
肠道疾病在代谢综合征中很常见。然而，其发病机制尚不完全清楚。猪和人在相关代谢过程方面非常相似。在这里，我们成功地通过 CRISPR/Cas9 技术敲入三个人类疾病风险基因，构建了一种代谢疾病易感转基因（TG）巴马猪模型，以评估其作为人类肠道疾病模型的潜力，并探讨可能涉及的病理机制。我们发现，在高脂肪高蔗糖饮食（HFHSD）处理后，TG 猪的空肠屏障完整性受到破坏，炎症细胞浸润增加。我们发现 TG 猪空肠的转录组、相关微生物组谱和微生物代谢物短链脂肪酸（SCFA）含量存在显著差异。值得注意的是，我们发现 SLC26A3 在 TG 猪中显著下调。在 IPEC-J2 细胞中敲低或过表达 SLC26A3 基因显著影响 MUC2、MUC13 和 occludin 的表达。此外，体外实验进一步验证了 CDX2 直接调控 SLC26A3 的表达。在机制上，CDX2 通过结合其-1120 至-1070bp 启动子区域增强 SLC26A3 的表达，从而介导肠道屏障功能。TG 猪代表了一种有前途的模型，为与代谢紊乱相关的人类肠道代谢疾病的临床前研究提供了新的见解，并表明 SLC26A3 可能是肠道代谢疾病的潜在治疗靶点。
