MiR-128 Participates in the Pathogenesis of Slow Transit Constipation by Regulating the P38α/M-CSF Inflammatory Signaling Pathway

Abstract

Background: Slow transit constipation (STC) is a gastrointestinal disorder that adversely affects the quality of life. MicroRNAs are involved in the pathogenesis of functional gastrointestinal disorders. This study aims to investigate the molecular mechanism of miR-128 in STC. Methods: A murine model of STC was constructed based on morphine and rhubarb. MiRNA array, bioinformatic analysis and experimental validation were applied to confirm the miR-128 and its target signaling pathway. The expression of miR-128 and downstream genes analysis were performed in the models. Inflammatory cytokine release in response to miR-128 regulation was evaluated in vitro. Findings: The expression of stem cell factor (SCF) and neuron-specific enolase (NSE) were low in the STC model. In addition, STC group showed a significant downregulation of miR-128 and upregulation of p38α and macrophage colony stimulating factors (M-CSF). Moreover, we observed elevated inflammatory cytokine and decreased anti-inflammatory cytokine levels in colonic tissues. Furthermore, co-culture assays indicated that regulating expression of miR-128 in colonic epithelial cells induced the secretion of IL-6 and TNF-α by macrophages. Interpretation: Our study demonstrated that miR-128 regulated the p38α/M-CSF signaling pathway to promote chronic inflammatory responses and changes in the immune microenvironment of the colon, thereby offering potential insights into the pathogenesis of STC and therapeutic targets for its treatment. Funding Information: This work was supported by grants from the National Natural Science Foundation of China (No.81570492, No.81500505), Science and Technology Innovation Fostering Foundation of Zhongnan Hospital, Wuhan University (cxpy2017018), Medical Science and Technology Innovation Platform of Health Commission of Hubei Province /Zhongnan Hospital of Wuhan University (No.PTXM2019011), Health Commission of Hubei Province (No. WJ2019M206), Science and Technology Department of Hubei Province (No.2018CKB913), and the Clinical Research Special Fund of Wu Jieping Medical Foundation (No.320.6750.18467, No.320.6750.19089-14). Declaration of Interests: The authors declare no conflict of interests. Ethics Approval Statement: Experiments were performed under a project license (No.2018039) granted by the ethics committee of Zhongnan Hospital of Wuhan University, Wuhan, China, in compliance with Animal Research Center of Wuhan University guidelines for the care and use of animals.