Serum Exosomes Derived from Irritable Bowel Syndrome Patient Increase Cell Permeability via Regulating miR-148b-5p/RGS2 Signaling in Human Colonic Epithelium Cells

Abstract

Aim. Irritable bowel syndrome (IBS) is a multifactorial functional bowel disorder characterized by disruption of the intestinal barrier. Circulating exosomal microRNAs (miRNAs) are involved in regulating epithelial barrier function, and upregulation of miR-148b-5p has been detected in IBS. However, whether exosomal miR-148-5p is involved in the IBS pathogenesis remains unclear. This study was aimed at investigating the relationship of exosomal miR-148-5p with colonic epithelial permeability. Methods. Exosomes were isolated from the serum of IBS patients and healthy controls. HT-29 cells were cultured with the IBS-derived serum exosomes (IBS-exo). Exosome uptake assay was used to evaluate whether the IBS-exo could be absorbed by HT-29 cells. FITC-Dextran flux and transepithelial/endothelial electrical resistance were measured to evaluate epithelial permeability. A luciferase reporter assay was used to determine whether the regulator of G protein signaling- (RGS-) 2 is a target gene of miR-148b-5p. Results. miR-148b-5p was obviously elevated in the IBS-exo compared to the control-exo. Upregulation of miR-148b-5p was observed in the HT-29 cells cultured with IBS-exo. Exposure to IBS-exo increased cell permeability and decreased RGS2 expression. The IBS-exo-induced alterations were obviously reversed by interfering with the miR-148b-5p expression. Mimicking the IBS-exo treatment, miR-148b-5p overexpression increased cell permeability and downregulated RGS2 expression, which were abrogated by overexpressing RGS2. The luciferase reporter assay revealed that RGS2 was a direct target of miR-148b-5p. Conclusions. Serum-derived exosomes from IBS patients increase colonic epithelial permeability via miR-148b-5p/RGS2 signaling.

Exploring the clinical significance of miR‐148 expression variations in distinct subtypes of irritable bowel syndrome

“…The miR‐148 family plays a critical role in the regulation of IBS and other bowel diseases. For instance, the upregulation of miR‐148b‐5p in the exosome‐derived serum of IBS patients has been linked to increased colonic epithelial permeability, thereby exacerbating IBS symptoms (Xing et al., 2021). Moreover, the loss of individual miR‐148 family members or the entire family has been associated with disruptions in intestinal barrier function, thus aggravating colitis and colitis‐associated colon cancer (Tang et al., 2022).…”

Role of microRNAs in Disorders of Gut–Brain Interactions: Clinical Insights and Therapeutic Alternatives

“…ARF4 suppression in the IEC line increased the expression of tight junction proteins and decreased intestinal epithelial permeability [72]. Additionally HT-29 cells cultured with IBS-derived serum exosomes were found to have increased cell permeability and miR-148b expression, which led to downregulation of the regulator of G-protein signaling 2 (RGS2) expression [73]. These findings supported the crucial role of miRNAs in the development of impaired intestinal barrier function, which is a core pathophysiological mechanism in DGBIs [30].…”

Integrated Analysis of lncRNA-miRNA-mRNA ceRNA Network in Human Diarrhea Irritable Bowel Syndrome

“…QiQi Zhou [25] miR-127 down NF-κB in ammatory K Zhao [11] miR-145, miR-148-5p and miR-592 up IL-6, IL-10 and TNF-α in ammatory Damian Jacenik [26] miR-148b-5p up RGS2 intestinal barrier Ying Xing [14] hsa-miR-106b, hsa-miR-26a, hsa-miR-29b up MAKP not described Wenhua Tao 2016 [27] miR-510 down PRDX1 in ammatory Yu Zhang 2019 [28] [11] H19 down AQP1 and AQP3…”

“…In recent years,researchers have found that the remaining 98% of the genome that was once considered as nonfunctional "junk" includes noncoding RNAs (ncRNAs) that play important roles in a wide range of biological processes such as growth,development, and organ function. Non-coding RNAs have been classi ed as small non-coding RNAs, including microRNAs,and long non-coding RNAs (lncRNAs) [9] .CHAO et al [10] found that the down-regulation of LncRNA H19 resulted in the expression changes of AQP1 and AQP3 may play an important role in the occurrence and development of IBS-D.ZHAO et al [11] con rmed lncRNA TUG1 attenuated TNF-α-caused apoptosis and in ammatory response in ICC by down-regulating miR-127 and then inactivating NF-κB and Notch pathways in patients with IBS-D.Meanwhile,It has been proved that microRNAs (miRs) have been veri ed to be related with the progression and development of IBS-D [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28] .Such as hsa-miR-150 and hsa-miR-342-3p link to pain and in ammatory pathways both of which are thought to be dysregulated in IBS [12] ;The cxolonic mucosal microRNAs, microRNA-219a-5p and microRNA-338-3p are downregulated in Irritable Bowel Syndrome and are associated with barrier function and MAPK signaling [13] ;Serum exosomes derived from Irritable Bowel Syndrome patient increase cell permeability via regulating miR-148b-5p/RGS2 signaling in human colonic epithelium cells [14] .To sum up, lncRNA-miRNA-mRNA regulatory network exerts a vital part in IBS-D genesis and progression.At the same time, there are few reports about the ceRNA regulation mechanism of lncRNA-miRNA related to IBS-D and the interaction between ncRNAs.…”