Department of Epidemiology & Biostatistics, College of Public Health, Zhengzhou University, Zhengzhou, China.
Biochem Biophys Res Commun. 2012 Sep 14;426(1):135-40. doi: 10.1016/j.bbrc.2012.08.051. Epub 2012 Aug 17.
Epithelial barrier dysfunction plays a critical role in the pathogenesis of allergic diseases; the mechanism is to be further understood. The ubiquitin E3 ligase A20 (A20) plays a role in the endocytic protein degradation in the cells. This study aims to elucidate the role of A20 in the maintenance of gut epithelial barrier function.
Gut epithelial cell line, HT-29 cell, was cultured into monolayers to evaluate the barrier function in transwells. RNA interference was employed to knock down the A20 gene in HT-29 cells to test the role of A20 in the maintenance of epithelial barrier function. Probiotic derived proteins were extracted from the culture supernatants using to enhance the expression of A20 in HT-29 cells.
The results showed that the knockdown of A20 compromised the epithelial barrier function in HT-29 monolayers, mainly increased the intracellular permeability. The fusion of endosome/lysosome was disturbed in the A20-deficient HT-29 cells. Allergens collected from the transwell basal chambers of A20-deficient HT-29 monolayers still conserved functional antigenicity. Treating with probiotic derived proteins increased the expression of A20 in HT-29 cells and promote the barrier function.
A20 plays an important role in the maintenance of epithelial barrier function as shown by HT-29 monolayer. Probiotic derived protein increases the expression of A20 and promote the HT-29 monolayer barrier function.
上皮屏障功能障碍在过敏疾病的发病机制中起着关键作用;其机制尚待进一步阐明。泛素 E3 连接酶 A20(A20)在细胞内的内吞蛋白降解中起作用。本研究旨在阐明 A20 在维持肠道上皮屏障功能中的作用。
培养肠道上皮细胞系 HT-29 细胞单层,在 Transwell 中评估屏障功能。采用 RNA 干扰技术敲低 HT-29 细胞中的 A20 基因,以测试 A20 在维持上皮屏障功能中的作用。从培养上清液中提取益生菌衍生蛋白,以增强 HT-29 细胞中 A20 的表达。
结果表明,A20 的敲低削弱了 HT-29 单层中的上皮屏障功能,主要增加了细胞内通透性。A20 缺陷的 HT-29 细胞中的内体/溶酶体融合受到干扰。从 A20 缺陷的 HT-29 单层 Transwell 基底室收集的过敏原仍然保持功能性抗原性。用益生菌衍生蛋白处理可增加 HT-29 细胞中 A20 的表达并促进屏障功能。
A20 在 HT-29 单层中维持上皮屏障功能中起着重要作用。益生菌衍生蛋白增加 A20 的表达并促进 HT-29 单层屏障功能。