Knockdown of lncRNA H19 inhibits abnormal differentiation of small intestinal epithelial cells in diabetic mice

Abstract

Diabetes mellitus (DM) comprises a group of metabolic diseases characterized by insulin deficiency or resistance and hyperglycemia. We previously reported the presence of abnormal differentiation of small intestinal epithelial cells (IECs) in diabetic mice, but the exact mechanism of this phenomenon has not been thoroughly elucidated to date. In this study, we found that H19 was markedly upregulated in IECs of DM mice. H19 knockdown significantly inhibited abnormal differentiation of IECs in DM mice. Bioinformatics analysis identified miR-141-3p as a candidate for H19. Based on luciferase reporter assays, we found that miR-141-3p directly targeted H19. Luciferase reporter assays also showed that miR-141-3p could directly target β-catenin. Furthermore, H19 might act as an endogenous "sponge" by competing for miR-141-3p binding to regulate miRNA targets in vitro and in vivo. In summary, our findings provide the first evidence supporting the role of H19 in IECs of DM mice, and miR-141-3p targets not only protein-coding genes but also the lncRNA H19.

Long noncoding RNA GAS5 inhibits cell proliferation and fibrosis in diabetic nephropathy by sponging miR-221 and modulating SIRT1 expression

“…lncRNA MIAT upregulates the TGFβ1 signaling pathway and enhances diabetic retinopathy [7]. In diabetic mice, the downregulation of lncRNA H19 suppresses the abnormal differentiation of small intestinal epithelial cells by competing with miR-141-3p [8]. The knockdown of HOTTIP regulates the cell cycle and insulin secretion by suppressing MEK/ERK axes in islet β cells [9].…”

Rspo3 regulates abnormal differentiation of small intestinal epithelial cells in diabetic state

“…Given the crucial roles in differentiation in IECs, examination of Rspo3 mechanism has provided new insight into the regulatory roles in stem cell behavior. Many studies have demonstrated miRNAs take part roles in differentiation of IECs in the mouse [14,25,26].Through miRNA expression pro les, bioinformatic analysis, and RT-qPCR, we noticed a miRNA, miR-380-5p, which was downregulated in IECs tissues. In addition, by an in situ hybridization of a DIG-labeled LNA-miR-380-5p probe, we found that distribution of miR-380-5p is consistent with Rspo3.…”

“…In our previous research, we found that the Wnt/β-catenin signaling pathway was continuously activated in IECs of DM mice [14], and the reason why Wnt/β-catenin pathway continuously was activated, and still need further study. In IESCs, the Wnt pathway is essential for intestinal crypt formation and renewal, whereas Rspo-mediated signalling mainly affects ISC numbers [20].…”

“…All mice received a tail vein injection once a day for 3 days. The db/+-NS group comprised control mice receiving saline (0.9%; same volume as the experimental group) injections [14,15,17,18], and the db/db-NS group comprised DM mice receiving saline (0.9%; same volume as the experimental group) injections [14,15,17,18]; the db/db-si-Rspo3 mice received injections of Rspo3 siRNA (80 mg/kg body weight, 14,15,17,18), and the db/db-agmiR-380-5p mice received injections of agmiR-380-5p (80 mg/kg body weight, 14,15,17,18). In each group, six mice were euthanized with an intraperitoneal injection of ketamine/xylazine (100/10 mg/kg body weight) on day 0 (prior to injection), day 2, day 4, and day 6 for further study.…”

“…The small intestine was dissected out from mice to expose the crypts and villi through sliced longitudinally. All procedures as described previously [12,14,15,16].…”