College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
Gansu Research Center for Swine Production Engineering and Technology, Lanzhou 730070, China.
Genes (Basel). 2023 May 6;14(5):1047. doi: 10.3390/genes14051047.
The () beta2 (CPB2) toxin produced by type C () can cause necrotizing enteritis in piglets. Immune system activation in response to inflammation and pathogen infection is aided by long non-coding RNAs (lncRNAs). In our previous work, we revealed the differential expression of the novel lncRNA LNC_001186 in -infected ileum versus healthy piglets. This implied that LNC_001186 may be a regulatory factor essential for infection in piglets. Herein, we analyzed the coding ability, chromosomal location and subcellular localization of LNC_001186 and explored its regulatory role in CPB2 toxin-induced apoptosis of porcine small intestinal epithelial (IPEC-J2) cells. RT-qPCR results indicated that LNC_001186 expression was highly enriched in the intestines of healthy piglets and significantly increased in -infected piglets' ileum tissue and CPB2 toxin-treated IPEC-J2 cells. The total sequence length of LNC_001186 was 1323 bp through RACE assay. CPC and CPAT, two online databases, both confirmed that LNC_001186 had a low coding ability. It was present on pig chromosome 3. Cytoplasmic and nuclear RNA isolation and RNA-FISH assays showed that LNC_001186 was present in the nucleus and cytoplasm of IPEC-J2 cells. Furthermore, six target genes of LNC_001186 were predicted using and approaches. Meanwhile, we constructed ceRNA regulatory networks with LNC_001186 as the center. Finally, LNC_001186 overexpression inhibited IPEC-J2 cells' apoptosis caused by CPB2 toxin and promoted cell viability. In summary, we determined the role of LNC_001186 in IPEC-J2 cells' apoptosis caused by CPB2 toxin, which assisted us in exploring the molecular mechanism of LNC_001186 in -induced diarrhea in piglets.
由 C 型产气荚膜梭菌（）产生的β2（CPB2）毒素可导致仔猪坏死性肠炎。免疫系统对炎症和病原体感染的激活，是由长链非编码 RNA（lncRNA）辅助的。在我们之前的工作中，我们揭示了新型 lncRNA LNC_001186 在感染肠段与健康仔猪之间的差异表达。这意味着 LNC_001186 可能是仔猪感染的关键调节因子。在此，我们分析了 LNC_001186 的编码能力、染色体位置和亚细胞定位，并探讨了其在 CPB2 毒素诱导猪小肠上皮细胞（IPEC-J2）凋亡中的调控作用。RT-qPCR 结果表明，LNC_001186 在健康仔猪的肠道中表达丰富，在感染仔猪的回肠组织和 CPB2 毒素处理的 IPEC-J2 细胞中显著增加。通过 RACE 试验，LNC_001186 的全长序列为 1323bp。两个在线数据库 CPC 和 CPAT 均证实 LNC_001186 具有较低的编码能力。它位于猪染色体 3 上。细胞质和核 RNA 分离和 RNA-FISH 试验表明，LNC_001186 存在于 IPEC-J2 细胞的核和细胞质中。此外，使用和 方法预测了 LNC_001186 的六个靶基因。同时，我们构建了以 LNC_001186 为中心的 ceRNA 调控网络。最后，LNC_001186 的过表达抑制了 CPB2 毒素引起的 IPEC-J2 细胞凋亡，提高了细胞活力。总之，我们确定了 LNC_001186 在 CPB2 毒素引起的 IPEC-J2 细胞凋亡中的作用，有助于我们探索 LNC_001186 在仔猪感染引起的腹泻中的分子机制。
