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

Abstract

Objective: The problems caused by diabetes mellitus (DM) related complications are the focus in clinical treatment. However, little is known about diabetic enteropathy (DE) and its the potential underlying mechanism. Methods: Intestinal cells (IEC) and Intestinal stem cells (IESC) obtained from BKS.Cg-Dock7m+/+Leprdb/JNju (DM) mice were used to detect Rspo3 by RT-qPCR, western blotting, Immunohistochemistry, and immunofluorescence. The role of Rspo3 in the abnormal differentiation of IECs of DM was clarified by knockout experiments. Through miRNA expression profiles, bioinformatic analysis, and RT-qPCR, we further analyzed differentiation related miRNA from IECs in DM mice. Results: The abnormal differentiation of small intestinal epithelial cells (IECs) was found in DM state. The expression of R-spondin 3 (Rspo3) was upregulated in IECs of DM state. And this phenomenon was associated with R-spondin 3 (Rspo3) overexpression. Additionally, Rspo3 is a major determinant of Lgr5+ stem cell identity upon DM state. Microarray analysis, Bioinformatics analysis and luciferase reporter assays revealed that microRNA (miR)-380-5p was directly targeted Rspo3. Moreover, miR-380-5p upregulation was observed to attenuate the abnormal differentiation of IECs through Rspo3 expression. Conclusion: Together, our results provide definitive evidence for the essential role of Rspo3 in differentiation of small intestinal epithelial cells (IECs) in DM state.