scholarly journals MiRNA-182-5p aggravates experimental ulcerative colitis via sponging Claudin-2

2021 ◽  
Author(s):  
Siwen Tang ◽  
Wentao Guo ◽  
Liumin Kang ◽  
Jinghua Liang
Keyword(s):  
Tight Junction ◽  
Gene Prediction ◽  
Luciferase Reporter Assay ◽  
Mucosal Barrier ◽  
Luciferase Reporter ◽  
Protective Effects ◽  
Reporter Assay ◽  
Tight Junction Proteins ◽  
Colitis Model ◽  
Junction Proteins

AbstractTight junction proteins play crucial roles in maintaining the integrity of intestinal mucosal barrier. MiRNA-182-5p is capable of targeting claudin-2 which is one of the vital tight junction proteins and the effect and mechanism of miRNA-182-5p was explored here in the DSS-induced colitis model. The pathological conditions were evaluated via hematoxylin and eosin staining. The gene expression level was assessed via PCR. Quantitative immunohistochemistry analysis was performed for the measurement of claudin-2. microRNA.org online tool was used for target gene prediction. Luciferase reporter assay and RNA pull-down assay were performed to detect the target of miRNA-182-5p. The inflammatory and oxidative stress level were measured using corresponding kits. MiRNA-182-5p was highly expressed in colitis model and miRNA-182-5p inhibitor exerted protective effects on colitis induced by DSS in mice. The protective effects includded improvement of pathological changes, increases in anti-inflammation and anti-oxidative genes, and up-regulation of TGF-β1. Claudin-2 mRNA was predicted as the target of miRNA-182-5p, which was validated via luciferase reporter assay and RNA pull-down assay. Claudin-2 overexpression was found in miRNA-182-5p inhibitor group. Consistent with the role of miRNA-182-5p, claudin-2 overexpression also exerted protective effects on DSS-induced colitis in mice. Inhibition of miRNA-182-5p exerted protective effects on colitis via targeting and upregulating claudin-2. The findings in study provide a new therapeutic strategy for colitis treatment and lay the foundation for future study.

scholarly journals Dietary l-Tryptophan Supplementation Enhances the Intestinal Mucosal Barrier Function in Weaned Piglets: Implication of Tryptophan-Metabolizing Microbiota

2018 ◽  
Vol 20 (1) ◽  
pp. 20 ◽  
Author(s):  
Haiwei Liang ◽  
Zhaolai Dai ◽  
Jiao Kou ◽  
Kaiji Sun ◽  
Jingqing Chen ◽  
...  
Keyword(s):  
Tight Junction ◽  
Mucosal Barrier ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Tight Junction Proteins ◽  
Weaned Piglets ◽  
Intestinal Mucosal Barrier ◽  
Mucosal Barrier Function ◽  
Small Intestinal ◽  
Junction Proteins

l-Tryptophan (Trp) is known to play an important role in the health of the large intestine. However, a role of dietary Trp in the small-intestinal mucosal barrier and microbiota remains poorly understood. The present study was conducted with weaned piglets to address this issue. Postweaning piglets were fed for 4 weeks a corn- and soybean meal-based diet supplemented with 0 (Control), 0.1, 0.2, or 0.4% Trp. The small-intestinal microbiota and serum amino acids were analyzed by bacterial 16S rRNA gene-based high-throughput sequencing methods and high-performance liquid chromatography, respectively. The mRNA levels for genes involved in host defense and the abundances of tight-junction proteins in jejunum and duodenum were measured by real time-PCR and Western blot techniques, respectively. The concentrations of Trp in the serum of Trp-supplemented piglets increased in a dose-dependent manner. Compared with the control group, dietary supplementation with 0.2–0.4% Trp reduced the abundances of Clostridium sensu stricto and Streptococcus in the jejunum, increased the abundances of Lactobacillus and Clostridium XI (two species of bacteria that can metabolize Trp) in the jejunum, and augmented the concentrations of secretory immunoglobulin A (sIgA) as well as mRNA levels for porcine β-defensins 2 and 3 in jejunal tissues. Moreover, dietary Trp supplementation activated the mammalian target of rapamycin signaling and increased the abundances of tight-junction proteins (zonula occludens (ZO)-1, ZO-3, and claudin-1) in jejunum and duodenum. We suggested that Trp-metabolizing bacteria in the small intestine of weaned pigs primarily mediated the beneficial effects of dietary Trp on its mucosal integrity, health, and function.

The Combined Effect of Vitamin C and Vitamin D3 on the Intestinal Epithelial Barrier by Regulating Notch Signaling Pathway

2019 ◽  
Author(s):  
Fubin Qiu ◽  
Zehui Zhang ◽  
Ying Ma ◽  
Linxue Yang ◽  
Rui Li
Keyword(s):  
Vitamin D ◽  
Vitamin C ◽  
Tight Junction ◽  
Mucosal Barrier ◽  
Control Group ◽  
High Dose ◽  
Tight Junction Proteins ◽  
Intestinal Mucosal Barrier ◽  
Notch Signal ◽  
Junction Proteins

Abstract Background: Tight junction proteins play crucial role in maintaining the intestinal mucosal barrier. Although previous studies had shown that the Notch signal is closely related to tight junction proteins, the mechanism by which it does so remains unknown. The goal of the present study was to investigate whether vitamin C combined with vitamin D3 affects intestinal mucosal barrier stability through Notch signal pathway.Results: To assess the effect of vitamin C combined with vitamin D3 on the intestinal mucosal barrier, electron microscopic observation of ultrastructure of tight junctions was done. And tight junction proteins gene and Notch signal gene expression were analyzed by quantitative reverse-transcription polymerase chain reaction, expression of tight junction protein in SW480 cells interfered with by LPS were examined by western blot. We found that vitamin C combined with vitamin D3 had protective effect on DSS-induced ulcerative colitis in guinea pig intestinal mucosa. Electron microscopy results showed that both low dose and high dose of vitamin C combined with vitamin D3 could maintain DSS-induced ulcerative colitis in guinea pig intestinal epithelium tight junction, however, the combination of medium dose vitamin C and vitamin D3 did not have this effect; Compared with the control group, the expression level of ZO-1 mRNA in the colon tissue of high-dose vitamin C group was significantly increased. In SW480 cell experiments, compared with the control group, the cell migration and repair ability of different concentrations of vitamin C combined with vitamin D3 group were significantly improved, the protein expression of Notch-1 was increased, but the protein expression of claudin-2 was significantly decreased. Conclusions: our results of this experiment showed that the appropriate amount of vitamin C combined with vitamin D3 might regulate the expression of claudin-2 by regulating Notch-1, slow the intestinal mucosal barrier destruction, and promote the damage repair of cell mucosal barrier.

scholarly journals Formononetin Administration Ameliorates Dextran Sulfate Sodium-Induced Acute Colitis by Inhibiting NLRP3 Inflammasome Signaling Pathway

2018 ◽  
Vol 2018 ◽  
pp. 1-12 ◽  
Author(s):  
Dacheng Wu ◽  
Keyan Wu ◽  
Qingtian Zhu ◽  
Weiming Xiao ◽  
Qing Shan ◽  
...  
Keyword(s):  
Tight Junction ◽  
Nlrp3 Inflammasome ◽  
Dextran Sulfate ◽  
Dextran Sulfate Sodium ◽  
Acute Injury ◽  
Dependent Manner ◽  
Protective Effects ◽  
Tight Junction Proteins ◽  
Acute Colitis ◽  
Junction Proteins

Formononetin is a kind of isoflavone compound and has been reported to possess anti-inflammatory properties. In this present study, we aimed to explore the protective effects of formononetin on dextran sulfate sodium- (DSS-) induced acute colitis. By intraperitoneal injection of formononetin in mice, the disease severity of colitis was attenuated in a dose-dependent manner, mainly manifesting as relieved clinical symptoms of colitis, mitigated colonic epithelial cell injury, and upregulations of colonic tight junction proteins levels (ZO-1, claudin-1, and occludin). Meanwhile, our study found that formononetin significantly prevented acute injury of colonic cells induced by TNF-α in vitro, specifically manifesting as the increased expressions of colonic tight junction proteins (ZO-1, claudin-1, and occludin). In addition, the result showed that formononetin could reduce the NLRP3 pathway protein levels (NLRP3, ASC, IL-1β) in vivo and vitro, and MCC950, the NLRP3 specific inhibitor, could alleviate the DSS-induced mice acute colitis. Furthermore, in the foundation of administrating MCC950 to inhibit activation of NLRP3 inflammasome, we failed to observe the protective effects of formononetin on acute colitis in mice. Collectively, our study for the first time confirmed the protective effects of formononetin on DSS-induced acute colitis via inhibiting the NLRP3 inflammasome pathway activation.

Protective effects of Notoginsenoside R1 on intestinal ischemia-reperfusion injury in rats

2014 ◽  
Vol 306 (2) ◽  
pp. G111-G122 ◽  
Author(s):  
Chong Li ◽  
Quan Li ◽  
Yu-Ying Liu ◽  
Ming-Xia Wang ◽  
Chun-Shui Pan ◽  
...  
Keyword(s):  
Tight Junction ◽  
Intestinal Ischemia ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Clinical Problem ◽  
Protective Effects ◽  
Tight Junction Proteins ◽  
Sprague Dawley ◽  
Component Form ◽  
Junction Proteins

Intestinal ischemia and reperfusion (I/R) is a clinical problem occurred for diverse causes with high mortality. Prophylaxis and treatment of intestinal I/R remains a challenge for clinicians. The purpose of the present study was to explore the role of Notoginsenoside R1 (R1), a major component form of Panax notoginseng, in management of intestinal I/R injury. Intestinal I/R was induced in male Sprague-Dawley rats by clamping the superior mesenteric artery for 90 min followed by reperfusion for 60 min or 3 days. R1 (10 mg·kg−1·h−1) was administered either 20 min before ischemia or 20 min after reperfusion. Intestinal microcirculation was evaluated by intravital microscopy over 60 min reperfusion. Sixty minutes or 3 days after reperfusion, rats were killed for histological examination of the jejunum tissue and immunohistochemical localization of myeloperoxidase and CD68. ATP, ADP, and AMP content in jejunum tissue was assessed by ELISA. Activation of nuclear factor-κB (NF-κB) and expression of ATP5D and tight junction proteins were determined by Western blotting. The results demonstrated that R1 is capable of attenuating intestinal I/R-induced microvascular hyperpermeability, inflammatory cytokine production, NF-κB activation, and loss of tight junction proteins, as well as improving energy metabolism during I/R. The results of the present study suggest R1 as an option in protecting against intestinal I/R injury.

scholarly journals Colquhounia Root Tablet Protects Rat Pulmonary Microvascular Endothelial Cells against TNF-α-Induced Injury by Upregulating the Expression of Tight Junction Proteins Claudin-5 and ZO-1

2018 ◽  
Vol 2018 ◽  
pp. 1-11 ◽  
Author(s):  
Wenjie Zhou ◽  
Guocui Shi ◽  
Jijia Bai ◽  
Shenmao Ma ◽  
Qinfu Liu ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Tight Junction ◽  
Cell Injury ◽  
Inflammatory Responses ◽  
Microvascular Endothelial Cells ◽  
Protective Effects ◽  
Tight Junction Proteins ◽  
Pulmonary Microvascular Endothelial Cells ◽  
Microvascular Endothelial ◽  
Junction Proteins

Background. There are currently limited effective pharmacotherapy agents for acute lung injury (ALI). Inflammatory response in the lungs is the main pathophysiological process of ALI. Our preliminary data have shown that colquhounia root tablet (CRT), a natural herbal medicine, alleviates the pulmonary inflammatory responses and edema in a rat model with oleic acid-induced ALI. However, the potential molecular action mechanisms underlining its protective effects against ALI are poorly understood. This study aimed to investigate the effects and mechanism of CRT in rat pulmonary microvascular endothelial cells (PMEC) with TNF-α-induced injury. Methods. PMECs were divided into 6 groups: normal control, TNF-α (10 ng/mL TNF-α), Dex (1×10-6 M Dex + 10 ng/mL TNF-α), CRT high (1000 ng/mL CRT + 10 ng/mL TNF-α), CRT medium (500 ng/mL CRT + 10 ng/mL TNF-α), and CRT low group (250 ng/mL CRT + 10 ng/mL TNF-α). Cell proliferation and apoptosis were detected by MTT assay and flow cytometry. Cell micromorphology was observed under transmission electron microscope. The localization and expression of tight junction proteins Claudin-5 and ZO-1 were analyzed by immunofluorescence staining and Western blot, respectively. Results. TNF-a had successfully induced an acute endothelial cell injury model. Dex and CRT treatments had significantly stimulated the growth and reduced the apoptosis of PMECs (all p < 0.05 or 0.01) and alleviated the TNF-α-induced cell injury. The expression of Claudin-5 and ZO-1 in Dex and all 3 CRT groups was markedly increased compared with TNF-a group (all p < 0.05 or 0.01). Conclusion. CRT effectively protects PMECs from TNF-α-induced injury, which might be mediated via stabilizing the structure of tight junction. CRT might be a promising, effective, and safe therapeutic agent for the treatment of ALI.

scholarly journals Down-Regulation of lncRNA XIST Ameliorates Lipoprotein(a) Induced Endothelial Progenitor Cell Damage via Modulating miR-126/PLK2 Axis

2021 ◽  
Author(s):  
Xiaolei Zhang ◽  
Guoying Xu ◽  
Shizhen Wang ◽  
Pei Chen ◽  
Shanshan Chen
Keyword(s):  
Cell Damage ◽  
Luciferase Reporter Assay ◽  
Luciferase Reporter ◽  
Protective Effects ◽  
Reporter Assay ◽  
Endothelial Progenitor ◽  
Potential Binding ◽  
Lncrna Xist ◽  
Induced Apoptosis ◽  
Dual Luciferase Reporter Assay

Abstract Background Recent studies have discovered that lncRNAs regulate lipoprotein (LP) (a)-mediated endothelial progenitor cells (EPCs) damage through sponging miRNAs. However, the role of XIST in EPC remains unknown. Method: Firstly, LP(a)(100 ng/mL) was used to treat EPCs for 6 hours. Then, the apoptosis, proliferation, migration, adhesion, angiogenesis of EPCs were detected. Moreover, overexpression of XIST or miR-126 on EPCs were built. Mechanistically, bioinformatics database, dual-luciferase reporter assay and RIP were used to found the binding relationships between XIST and miR-126, miR-126 and PLK2. Results Presently, we found LP(a) treatment significantly induced apoptosis, attenuated the proliferation, migration, adhesion, angiogenesis of EPCs. While knocking down XIST or overexpression miR-126 significantly reversed LP(a) induced damage on EPCs. Moreover, forced overexpression of XIST partially offset the protective effects of overexpressing miR-126 on EPC. Mechanistically, through bioinformatics database (http://starbase.sysu.edu.cn/index.php), we found potential binding relationships between XIST and miR-126, miR-126 and PLK2. Furthermore, the dual-luciferase reporter assay and RIP experiment confirmed the targeted binding between them. Conclusion Collectively, the above results confirmed that down-regulating XIST improved LP(a)-induced EPC damage by regulating the miR-126/PLK2 axis, and the XIST/miR-126/PLK2 axis exerted an essential role in regulating EPC function.

Filaggrin and Tight Junction Proteins are Crucial for Esophageal Mucosal Barrier in Eosinophilic Esophagitis

2017 ◽  
Vol 152 (5) ◽  
pp. S853-S854
Author(s):  
Tadayuki Oshima ◽  
Liping Wu ◽  
Masato Taki ◽  
Toshihiko Tomita ◽  
Yoshio Ohda ◽  
...  
Keyword(s):  
Tight Junction ◽  
Eosinophilic Esophagitis ◽  
Mucosal Barrier ◽  
Tight Junction Proteins ◽  
Junction Proteins

scholarly journals miR-184 Inhibits Tumor Invasion, Migration and Metastasis in Nasopharyngeal Carcinoma by Targeting Notch2

2018 ◽  
Vol 49 (4) ◽  
pp. 1564-1576 ◽  
Author(s):  
Hong-Ming Zhu ◽  
Xue-Song Jiang ◽  
Hui-Zi Li ◽  
Lu-Xi Qian ◽  
Ming-Yu Du ◽  
...  
Keyword(s):  
Nasopharyngeal Carcinoma ◽  
Cell Invasion ◽  
Mirna Target ◽  
Target Gene ◽  
Gene Prediction ◽  
Luciferase Reporter Assay ◽  
Luciferase Reporter ◽  
Reporter Assay ◽  
Mirna Target Gene ◽  
Dual Luciferase Reporter Assay

Background/Aims: A recent study found that dysregulated microRNA-184 (miR-184) is involved in the proliferation and survival of nasopharyngeal carcinoma (NPC). This study aimed to evaluate the detailed mechanisms of invasion, migration and metastasis of NPC cells. Methods: Quantitative reverse-transcription PCR (qRT-PCR) and Western blot were used to confirm the expression levels of miR-184 and Notch2. NPC cell invasion and migration were subsequently examined using in vitro cell invasion and wound-healing assays, respectively. MicroRNA (miRNA) target gene prediction databases and dual-luciferase reporter assay were adopted to validate the target genes of miR-184. Results: MiR-184 was downregulated in the NPC cell lines. The miR-184 inhibitor increased the number of invading NPC cells, whereas miR-184 mimics inhibited the invasive ability of such cells. The protein level of E-cadherin decreased, whereas those of N-cadherin and vimentin increased in the anti-miR-184 group. This result showed that miR-184 inhibited NPC cell invasion and metastasis by regulating EMT progression. MiRNA target gene prediction databases indicated the potential of Notch2 as a direct target gene of miR-184. Such a notion was then validated by results of dual-luciferase reporter assay. Notably, shRNANotch2 restrained the EMT and partially abrogated the inhibitory effects of miR-184 on EMT progression in NPC cells. Conclusion: MiR-184 functions as a tumour-suppressive miRNA targeting Notch2 and inhibits the invasion, migration and metastasis of NPC.

Sign in / Sign up

Export Citation Format

Share Document