Department of Medicine, Graduate School of Medicine, Shiga University of Medical Science, Seta Tsukinowa-cho, Otsu, Shiga 520-2192, Japan.
Mol Med Rep. 2011 May-Jun;4(3):483-7. doi: 10.3892/mmr.2011.442. Epub 2011 Feb 18.
Interleukin (IL)-32 plays a role in the pathophysiology of inflammatory bowel disease (IBD). We isolated a new isoform of the IL-32 transcript in the process of cloning the full-length IL-32 gene from human colonic subepithelial myofibroblasts (SEMFs). The expression of mRNA in the samples was assessed by RT-PCR and real-time PCR analyses. The PCR products from the IL-32 genes were ligated into the expression vector pIRESneo2. The new isoform of the IL-32 transcript (336 nucleotides) completely lacked exon 4 of the IL-32γ gene, and was 60 bp shorter than IL-32α. TNF-α induced the mRNA expression of the new IL-32 isoform in a dose- and time-dependent manner. Stable transfection of this new isoform significantly decreased TNF-α-induced IL-8 mRNA expression in HT-29 cells, but the expression of the IL-32α gene had no effect. The mRNA expression of this new isoform was significantly elevated in the inflamed mucosa of IBD patients. A new isoform of the IL-32 transcript may play an anti-inflammatory role in the inflamed mucosa of IBD.
白细胞介素 (IL)-32 在炎症性肠病 (IBD) 的病理生理学中发挥作用。在从人结肠黏膜下肌成纤维细胞 (SEMF) 克隆全长 IL-32 基因的过程中，我们分离到了 IL-32 转录本的一种新同工型。通过 RT-PCR 和实时 PCR 分析评估样本中的 mRNA 表达。将 IL-32 基因的 PCR 产物连接到表达载体 pIRESneo2 中。新的 IL-32 转录本同工型（336 个核苷酸）完全缺乏 IL-32γ 基因的外显子 4，比 IL-32α 短 60 个碱基。TNF-α 以剂量和时间依赖的方式诱导新的 IL-32 同工型的 mRNA 表达。该新同工型的稳定转染可显著降低 HT-29 细胞中 TNF-α 诱导的 IL-8 mRNA 表达，但 IL-32α 基因的表达没有影响。该新同工型的 mRNA 表达在 IBD 患者的炎症黏膜中显著升高。IL-32 转录本的一种新同工型可能在 IBD 的炎症黏膜中发挥抗炎作用。
