■瞬时受体电位锚蛋白1(TRPA1)通道在肠道固有层中高表达,但其对肠道生理学/病理生理学的贡献尚不清楚。这里,我们评估了肌成纤维细胞TRPA1通道在肠道重塑中的功能。
■通过转化生长因子-β1刺激肠成肌纤维细胞系(InMyoFibs)以诱导体外纤维化。使用簇状规则间隔短回文重复(CRISPR)/CRISPR相关9(Cas9)系统产生Trpal敲除小鼠。通过每周直肠内给药三硝基苯磺酸(TNBS)建立小鼠慢性结肠炎模型。来自克罗恩病(CD)患者肠道的样品用于病理染色和定量分析。
■在InMyoFibs中,TRPA1在TRP家族成员中表达量最高。在TNBS慢性结肠炎模型小鼠中,与野生型小鼠相比,TRPA1-/-基因敲除小鼠的炎症和纤维化改变程度更为显著.在野生型小鼠中给予泼尼松龙抑制纤维化病变的一周灌肠,但不是在TRPA1基因敲除小鼠中。类固醇和吡非尼酮诱导InMyoFibs中的Ca2流入,被选择性TRPA1通道阻断剂HC-030031拮抗。类固醇和吡非尼酮可抵抗转化生长因子β1诱导的热休克蛋白47,1型胶原的表达,和α-平滑肌肌动蛋白,并降低了InMyoFibs中Smad-2的磷酸化和myocardin的表达。在CD患者的狭窄肠道区域,TRPA1表达显著增高。TRPA1/热休克蛋白47双阳性细胞在CD患者和TNBS治疗小鼠的狭窄肠区积累。
■TRPA1,除了抗炎作用外,可以预防肠道纤维化,因此成为高度无法治愈的炎性/纤维化疾病的新治疗靶标。
UNASSIGNED: The transient receptor potential ankyrin 1 (TRPA1) channel is highly expressed in the intestinal lamina propria, but its contribution to gut physiology/pathophysiology is unclear. Here, we evaluated the function of myofibroblast TRPA1 channels in intestinal remodeling.
UNASSIGNED: An intestinal myofibroblast cell line (InMyoFibs) was stimulated by transforming growth factor-β1 to induce in vitro fibrosis. Trpa1 knockout mice were generated using the Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) system. A murine chronic colitis model was established by weekly intrarectal trinitrobenzene sulfonic acid (TNBS) administration. Samples from the intestines of Crohn\'s disease (CD) patients were used for pathologic staining and quantitative analyses.
UNASSIGNED: In InMyoFibs, TRPA1 showed the highest expression among TRP family members. In TNBS chronic colitis model mice, the extents of inflammation and fibrotic changes were more prominent in TRPA1-/- knockout than in wild-type mice. One-week enema administration of prednisolone suppressed fibrotic lesions in wild-type mice, but not in TRPA1 knockout mice. Steroids and pirfenidone induced Ca2+ influx in InMyoFibs, which was antagonized by the selective TRPA1 channel blocker HC-030031. Steroids and pirfenidone counteracted transforming growth factor-β1-induced expression of heat shock protein 47, type 1 collagen, and α-smooth muscle actin, and reduced Smad-2 phosphorylation and myocardin expression in InMyoFibs. In stenotic intestinal regions of CD patients, TRPA1 expression was increased significantly. TRPA1/heat shock protein 47 double-positive cells accumulated in the stenotic intestinal regions of both CD patients and TNBS-treated mice.
UNASSIGNED: TRPA1, in addition to its anti-inflammatory actions, may protect against intestinal fibrosis, thus being a novel therapeutic target for highly incurable inflammatory/fibrotic disorders.