MicroRNA-200a Targets Cannabinoid Receptor 1 and Serotonin Transporter to Increase Visceral Hyperalgesia in Diarrhea-predominant Irritable Bowel Syndrome Rats

Abstract

Background/AimsMicroRNAs (miRNAs) were reported to be responsible for intestinal permeability in diarrhea-predominant irritable bowel syndrome (IBS-D) rats in our previous study. However, whether and how miRNAs regulate visceral hypersensitivity in IBS-D remains largely unknown.MethodsWe established the IBS-D rat model and evaluated it using the nociceptive visceral hypersensitivity test, myeloperoxidase activity assay, restraint stress-induced defecation, and electromyographic (EMG) activity. The distal colon was subjected to miRNA microarray analysis followed by isolation and culture of colonic epithelial cells (CECs). Bioinformatic analysis and further experiments, including dual luciferase assays, quantitative real-time polymerase chain reaction, western blot, and enzyme-linked immunosorbent assay, were used to detect the expression of miRNAs and how it regulates visceral hypersensitivity in IBS-D rats.ResultsThe IBS-D rat model was successfully established. A total of 24 miRNAs were differentially expressed in the distal colon of IBS-D rats; 9 were upregulated and 15 were downregulated. Among them, the most significant upregulation was miR-200a, accompanied by downregulation of cannabinoid receptor 1 (CNR1) and serotonin transporter (SERT). MiR-200a mimic markedly inhibited the expression of CNR1/SERT. Bioinformatic analysis and luciferase assay confirmed that CNR1/SERT are direct targets of miR-200a. Rescue experiments that overexpressed CNR1/SERT significantly abolished the inhibitory effect of miR-200a on the IBS-D rats CECs.ConclusionsThis study suggests that miR-200a could induce visceral hyperalgesia by targeting the downregulation of CNR1 and SERT, aggravating or leading to the development and progression of IBS-D. MiR-200a may be a regulator of visceral hypersensitivity, which provides potential targets for the treatment of IBS-D.

Efficacy of Tong-Xie-Yao-Fang granule and its impact on whole transcriptome profiling in diarrhea-predominant irritable bowel syndrome patients: study protocol for a randomized controlled trial

“…Other literatures have reported that in IBS-D patients, miR-199 and miR-29 are respectively involved in visceral pain and intestinal permeability (34, 35). Our previous studies also have proved that miR-144 and miR-200 by binding to speci c target mRNA play vital roles in IBS-D rats (22,23). However, study on lncRNAs and circRNAs in IBS-D is as yet scarce.…”

“…More and more studies have given insights into the indispensable roles of ncRNAs played in cellular process (19), tumorigenesis (20), in ammatory regulation (21) and so on. Our previous studies also have reported that microRNA-144 and microRNA-200a are involved in intestinal permeability and visceral hyperalgesia separately by binding to speci c target mRNA in IBS-D rats (22,23). However, exploration about ncRNAs for IBS-D is still negligible.…”

Efficacy of Tong-Xie-Yao-Fang granule and its impact on whole transcriptome profiling in diarrhea-predominant irritable bowel syndrome patients: study protocol for a randomized controlled trial

“…Other literatures have reported that in IBS-D patients, miR-199 and miR-29 are respectively involved in visceral pain and intestinal permeability [ 34 , 35 ]. Our previous studies also have proved that miR-144 and miR-200 by binding to specific target mRNA play vital roles in IBS-D rats [ 22 , 23 ]. However, the study on lncRNAs and circRNAs in IBS-D is as yet scarce.…”

“…More and more studies have given insights into the indispensable roles of ncRNAs played in the cellular process [ 19 ], tumorigenesis [ 20 ], inflammatory regulation [ 21 ], and so on. Our previous studies also have reported that microRNA-144 and microRNA-200a are involved in intestinal permeability and visceral hyperalgesia separately by binding to specific target mRNA in IBS-D rats [ 22 , 23 ]. However, exploration about ncRNAs for IBS-D is still negligible.…”

microRNA‐495 reduces visceral sensitivity in mice with diarrhea‐predominant irritable bowel syndrome through suppression of the PI3K/AKT signaling pathway via PKIB

“…The abnormal increase of visceral sensitivity may cause gastrointestinal dysfunction, and visceral hypersensitivity is thought to be an important pathophysiological factor . Diarrhea‐predominant IBS (IBS‐D), the largest sub‐category of IBS, can cause visceral pain, changed bowel habits, and bloating under normal biochemistry or anatomy . The treatments of IBS‐D include fiber supplements, probiotics, antidepressants, and a 5‐hydroxytryptophan 3 receptor antagonist, but those treatments are not so effective .…”