TRPA1 protects mice from pathogenic Citrobacter rodentium infection via maintaining the colonic epithelial barrier function
TRPA1 protects mice from pathogenic Citrobacter rodentium infection via maintaining the colonic epithelial barrier function
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TRPA1 通过维持结肠上皮屏障功能保护小鼠免受致病性柠檬酸杆菌感染
DOI：
10.1096/fj.202200483rrr
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发表时间：
2023-02-01
期刊：
影响因子：
4.8
通讯作者：
Lan, Lei
中科院分区：
文献类型：
--
作者：
Liu, Lin;Xu, Min;Lan, Lei
瞬时受体电位锚蛋白 1 (TRPA1) 在胃肠道中表达，在肠道蠕动和内脏超敏反应中发挥重要作用。然而，TRPA1 在宿主防御（特别是针对肠道病原体）中的潜在作用尚不清楚。在这里，我们发现 Trpa1 基因敲除小鼠表现出对柠檬酸杆菌感染的易感性增加，这与腹泻严重程度增加以及结肠上皮紧密连接 (TJ) 破坏相关的肠道通透性增加有关。我们进一步证明了 TRPA1 在小鼠结肠上皮细胞 (CEC) 和人上皮 Caco-2 细胞中在蛋白质水平和转录水平上的表达。使用钙成像，观察到 TRPA1 激动剂异硫氰酸烯丙酯 (AITC) 和过氧化氢分别在 Caco-2 细胞中诱导瞬时 Ca2+ 反应。此外，Caco-2细胞中TRPA1敲除导致TJ蛋白、ZO-1和Occludin表达降低，体外Caco-2单层细胞旁细胞通透性增加和TEER值降低。此外，在汇合的 Caco-2 细胞中，HC-030031 对 TRPA1 的抑制导致 TJ 蛋白、ZO-1、Occludin 和 Claudin-3 的分布和表达发生改变，并加剧了细菌内毒素脂多糖 (LPS) 诱导的对这些 TJ 蛋白和肌动蛋白细胞骨架的损伤。相比之下，AITC 预处理在 LPS 攻击后恢复了汇合 Caco-2 细胞中这些 TJ 蛋白的分布和表达。我们的结果确定了 TRPA1 在宿主防御肠道细菌病原体中具有未被认识到的保护作用，其通过维持结肠上皮屏障功能，至少部分是通过保留 TJ 蛋白在 CEC 中的分布和表达。
Transient receptor potential ankyrin 1 (TRPA1) is expressed in gastrointestinal tract and plays important roles in intestinal motility and visceral hypersensitivity. However, the potential role of TRPA1 in host defense, particularly against intestinal pathogens, is unknown. Here, we show that Trpa1 knockout mice exhibited increased susceptibility to Citrobacter rodentium infection, associated with the increased severity of diarrhea and intestinal permeability associated with the disrupted tight junctions (TJs) in colonic epithelia. We further demonstrated the expression of TRPA1 in murine colonic epithelial cells (CECs) and human epithelial Caco-2 cells both at protein level and transcription level. Using calcium imaging, TRPA1 agonists allyl isothiocyanates (AITC) and hydrogen peroxide were observed to induce a transient Ca2+ response in Caco-2 cells, respectively. Moreover, TRPA1 knockdown in Caco-2 cells resulted in the decreased expression of TJ proteins, ZO-1 and Occludin, and in the increased paracellular permeabilities and the reduced TEER values of Caco-2 monolayers in vitro. Furthermore, inhibition of TRPA1 by HC-030031 in the confluent Caco-2 cells caused the altered distribution and expression of TJ proteins, ZO-1, Occludin, and Claudin-3, and exacerbated the bacterial endotoxin lipopolysaccharide (LPS)-induced damage to these TJ proteins and actin cytoskeleton. By contrast, AITC pretreatment restored the distribution and expression of these TJ proteins in the confluent Caco-2 cells upon LPS challenge. Our results identify an unrecognized protective role of TRPA1 in host defense against an enteric bacterial pathogen by maintaining colonic epithelium barrier function, at least in part, via preserving the distribution and expression of TJ proteins in CECs.