Department of Gastroenterology, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510176, P.R. China.
School of Chinese Medicine, Hong Kong Baptist University, Hong Kong, China.
J Microbiol Biotechnol. 2020 Oct 28;30(10):1480-1487. doi: 10.4014/jmb.2002.02059.
Our previous report determined that miR-144 is a key regulator of intestinal epithelial permeability in irritable bowel syndrome with diarrhea (IBS-D) rats. Recent evidence has shown that lactobacilli play an important role in the relief of IBS-D symptoms. However, few studies have addressed the mechanisms by which microRNAs and lactobacilli exert their beneficial effects on intestinal epithelial permeability. Hence, to elucidate whether miRNAs and lactobacilli play roles in intestinal epithelial barrier regulation, we compared miRNA expression levels in intestinal epithelial cells (IECs) under ( LC01) treatment. IECs and LC01 were co-cultured and then subjected to microRNA microarray assay. qRT-PCR, western blot and ELISA were used to detect the expression of occludin (OCLN) and zonula occludens 1 (ZO1/TJP1). The interaction between miRNAs and LC01 acting in IECs was investigated through transfection of RNA oligoribonucleotides and pcDNA 3.1 plasmid. The results are as follows: 1) LC01 decreased the expression of miR-144 and FD4 and promoted OCLN and ZO1 expression in IECs; 2) LC01 enhanced the barrier function of IECs via downregulation of miR-144 and upregulation of OCLN and ZO1; 3) Under LC01 treatment, OCLN and ZO1 overexpression could partially eliminate the promoting effect of miR-144 on intestinal permeability in IECs. Our results demonstrate that LC01 regulates intestinal permeability of IECs through miR-144 targeting of OCLN and ZO1. LC01 can be a possible therapeutic target for managing dysfunction of the intestinal epithelial barrier.
我们之前的报告确定 miR-144 是腹泻型肠易激综合征（IBS-D）大鼠肠上皮通透性的关键调节因子。最近的证据表明，乳酸菌在缓解 IBS-D 症状方面发挥着重要作用。然而，很少有研究探讨 microRNAs 和乳酸菌发挥其对肠上皮通透性有益作用的机制。因此，为了阐明 microRNAs 和乳酸菌是否在肠上皮屏障调节中发挥作用，我们比较了 LC01 处理下肠上皮细胞（IECs）中的 miRNA 表达水平。将 IECs 和 LC01 共培养，然后进行 microRNA 微阵列分析。qRT-PCR、western blot 和 ELISA 用于检测 occludin（OCLN）和 zonula occludens 1（ZO1/TJP1）的表达。通过转染 RNA 寡核苷酸和 pcDNA 3.1 质粒研究了 miRNAs 和 LC01 在 IECs 中的相互作用。结果如下：1）LC01 降低了 miR-144 和 FD4 的表达，促进了 IECs 中 OCLN 和 ZO1 的表达；2）LC01 通过下调 miR-144 和上调 OCLN 和 ZO1 增强了 IECs 的屏障功能；3）在 LC01 处理下，OCLN 和 ZO1 的过表达可以部分消除 miR-144 对 IECs 肠通透性的促进作用。我们的结果表明，LC01 通过 miR-144 靶向 OCLN 和 ZO1 调节 IECs 的肠通透性。LC01 可能是管理肠上皮屏障功能障碍的潜在治疗靶点。
