College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
Gansu Research Center for Swine Production Engineering and Technology, Lanzhou 730070, China.
Int J Mol Sci. 2024 Oct 5;25(19):10715. doi: 10.3390/ijms251910715.
Inflammatory diseases of the intestinal tract in piglets severely impair the economic performance of pig farms. Pig milk exosomes can encapsulate miRNAs which can then enter the piglet intestine to play an immunomodulatory role. Previously, we comparatively analyzed and identified exosomal miRNAs in the colostrum and mature milk of Bamei and Landrace pigs, and we screened for ssc-miR-22-3p, which is associated with inflammation and immune response; however, the role played by ssc-miR-22-3p in the immune response in IPEC-J2 cells is not yet clear. In this study, we first constructed a pig intestinal inflammatory response model using Lipopolysaccharide (LPS) and Polyinosinic-polycytidylic acid (Poly (I:C)), and we investigated the role of ssc-miR-22-3p targeting in the regulation of LPS and Poly (I:C)-induced inflammatory injury in IPEC-J2 cells by RT-qPCR, cell counting kit-8 (CCK-8), EdU staining, lactate dehydrogenase (LDH) activity assay, and dual luciferase reporter gene assay. We successfully established LPS and Poly (I:C)-induced cell damage models in IPEC-J2 cells. The immune response of IPEC-J2 cells was stimulated by induction of IPEC-J2 cells at 10 μg/mL LPS and 20 μg/mL Poly (I:C) for 24 h. Overexpression of ssc-miR-22-3p decreased cytokine expression and promoted cell viability and proliferation. The functional enrichment analysis revealed that ssc-miR-22-3p targets genes enriched in the pathways of negative regulation of inflammatory response and bacterial invasion of epithelial cells. The validity of the binding site of ssc-miR-22-3p to was tested by a dual luciferase reporter gene. Pig milk exosome ssc-miR-22-3p promotes cell viability and proliferation by targeting , and it alleviates LPS and Poly (I:C)-induced inflammatory responses in IPEC-J2 cells.
仔猪肠道炎症性疾病严重损害猪场的经济效益。猪乳外泌体可以包裹 miRNA,然后进入仔猪肠道发挥免疫调节作用。之前,我们比较分析并鉴定了巴马猪和长白猪初乳和成熟乳中的外泌体 miRNA,并筛选出与炎症和免疫反应相关的 ssc-miR-22-3p;然而,ssc-miR-22-3p 在 IPEC-J2 细胞免疫反应中的作用尚不清楚。在本研究中,我们首先用脂多糖(LPS)和聚肌苷酸-聚胞苷酸(Poly(I:C))构建了猪肠道炎症反应模型,并用 RT-qPCR、细胞计数试剂盒-8(CCK-8)、EdU 染色、乳酸脱氢酶(LDH)活性测定和双荧光素酶报告基因测定研究了 ssc-miR-22-3p 靶向在 LPS 和 Poly(I:C)诱导的 IPEC-J2 细胞炎症损伤中的调节作用。我们成功建立了 LPS 和 Poly(I:C)诱导的 IPEC-J2 细胞损伤模型。用 10μg/ml LPS 和 20μg/ml Poly(I:C)诱导 IPEC-J2 细胞 24 h 刺激 IPEC-J2 细胞免疫反应。ssc-miR-22-3p 的过表达降低了细胞因子的表达,促进了细胞活力和增殖。功能富集分析表明,ssc-miR-22-3p 靶向的基因富集在炎症反应的负调节和细菌侵袭上皮细胞的途径中。双荧光素酶报告基因检测验证了 ssc-miR-22-3p 与 结合位点的有效性。猪乳外泌体 ssc-miR-22-3p 通过靶向 促进细胞活力和增殖,并减轻 LPS 和 Poly(I:C)诱导的 IPEC-J2 细胞炎症反应。