Expression Levels Of miRNA-146a Research Articles

Oleuropein as An Effective Suppressor of Inflammation and MicroRNA-146a Expression in Patients with Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a common progressive autoimmune disorder that causes chronic inflammation of the joints and damage to other organs. Previous studies have reported the important role of miRNA-146a in the pathogenesis of RA. In addition, the anti-inflammatory and modulatory effects of oleuropein (OLEU) on the expression pattern of microRNAs (miRNAs) have been shown in different diseases. Therefore, this study aimed to evaluate both the sensitivity and specificity of miRNA-146a and determine the potential effects of OLEU on the expression levels of miRNA-146a and tumour necrosis factor-alpha (TNF-α) in RA patients. The participants in this experimental study were divided into 2 groups: RA (n=45) and healthy controls (n=30). The isolated peripheral blood mononuclear cells (PBMCs) were treated with different concentrations of OLEU; and the level of TNF-α expression, anti-citrullinated protein, and miRNA-146a were determined using enzyme-linked immunoassay and real-time polymerase chain reaction, respectively. In addition, the receiver operating characteristic (ROC) curve analysis evaluated the sensitivity and specificity of miRNA-146a in RA patients. Results revealed a positive correlation between the levels of miRNA-146a expression with the serum levels of C-reactive protein (CRP) and rheumatoid factor (RF) in RA patients. In addition, OLEU treatment decreased the levels of TNF-α and miRNA-146a expression in treated PBMCs samples compared with untreated cells. The ROC curve analysis showed an 85% sensitivity and 100% specificity of miRNA-146a in RA patients. Therefore, miRNA-146a can be used as a useful biomarker for RA diagnosis, particularly for early detection. In addition, OLEU could suppress inflammation in RA patients through the regulation of miRNA-146a.

Associations of miRNA-146a and miRNA-223 with Rheumatoid Arthritis and Their Predictive Values.

To analyze the independent associations of miRNA-146a and miRNA-223 with rheumatoid arthritis (RA) and evaluate their predictive values for RA. A total of 68 RA patients were selected as cases, and meanwhile 68 patients with a traumatic knee condition were selected as controls by matching to the cases according to sex and age at the ratio of 1:1. The independent associations of miRNA-146a and miRNA-223 with RA were identified by binary logistic regression analysis. Receiver operating characteristic (ROC) curve was used to evaluate their predictive values for RA. MiRNA-146a and miRNA-223 expression levels in both synovial tissues and serums were statistically higher in cases than in controls, and their expression levels in serums were not statistically different from those in synovial tissues in both cases and controls. The expression levels of miRNA-146a and miRNA-223 in synovial tissues were independently associated with RA, as well as the expression levels of miRNA-146a and miRNA-223 in serums. The area under curve (AUC) of combination of miRNA-146a and miRNA-223 in synovial tissues for the prediction of RA was 0.910 [95% confidence interval (CI): 0.863-0.962], and the AUC of combination of miRNA-146a and miRNA-223 in serums was 0.904 (95% CI: 0.851-0.957). Their difference was not statistically significant (P=0.873), but the AUC of combination prediction was statistically higher than those of individual predictions (synovial tissues: 0.910 vs 0.773, P=0.005, 0.910 vs 0.788, P=0.009; serums: 0.904 vs 0.766, P=0.005, 0.904 vs 0.784, P=0.011). MiRNA-146a and miRNA-223 in both synovial tissues and serums could be applied in predicting RA, and their combination could elevate the predictive value significantly.

MiRNA-146a and miRNA-202-3p Attenuate Inflammatory Response by Inhibiting TLR4, IRAK1, and TRAF6 Expressions in Rats following Spinal Cord Injury

Spinal cord injury (SCI) is a catastrophic disease that induces a complex cascade of cellular reactions at the local lesion area, including secondary cell death and inflammatory reactions. Accumulating evidence has showed pro- and anti-inflammatory roles of microRNAs (miRNAs), a class of small RNAs, in SCI. The present study is aimed at investigating the effects of two miRNAs, miRNA-146a and miRNA-202-3p, on inflammatory response after SCI. Initially, we found that the expression levels of miRNA-146a and miRNA-202-3p were increased in the plasma samples of 32 SCI patients at days 3 and 7 after admission and the rat spinal cord at days 3 and 7 after SCI modeling compared with healthy controls and sham-operated rats, respectively. The expression levels of TLR4, IRAK1, and TRAF6 were declined in the rat spinal cord at days 1, 3, and 7 after SCI modeling compared with sham-operated rats. Injection of miRNA-146a mimic or miRNA-202-3p mimic decreased TLR4, IRAK1, and TRAF6 expressions in the rat spinal cord at days 1, 3, and 7 after SCI modeling, while injection of miRNA-146a antagomir or miRNA-202-3p antagomir produced opposed results. Subsequent results showed that the expression levels of tumor necrosis factor-α (TNF-α), IL-1β, IL-6, and IL-8 were upregulated in the rat serum at days 1, 3, and 7 after SCI modeling compared with sham-operated rats. Injection of miRNA-146a mimic or miRNA-202-3p mimic decreased TNF-α, IL-1β, IL-6, and IL-8 expression levels in the rat serum at days 1, 3, and 7 after SCI modeling, while injection of miRNA-146a antagomir or miRNA-202-3p antagomir yielded opposed results. The expression levels of TNF-α, IL-1β, IL-6, and IL-8 were higher in the supernatants of PC12 cells transfected with anti-miRNA-146a or anti-miRNA-202-3p than in those transfected with si-TLR4, si-IRAK1, or si-TRAF6. These findings support the notion that miRNA-146a/miRNA-202-3p exerts anti-inflammatory functions after SCI.

МикроРНК и эндокринная офтальмопатия, участие в глюкокортикоидном ответе

The accumulated clinical experience proves the existence of glucocorticoid-resistant forms of endocrine ophthalmopathy. It is assumed that epigenetic factors (miRNA) affect the pathogenesis of the disease and are involved in the formation of the glucocorticoid response. Purpose. To study the role of miRNA-146a and miRNA-155 in patients with clinical forms of endocrine ophthalmopathy and to assess their effect on the glucocorticoid response. Material and method: The study included 20 patients (40 orbits) with image intensifier tubes, treated with glucocorticoids (GC) with a negative result, mean age – 52.65 ± 12.6 years. The control group - 28 volunteers without autoimmune and tumor diseases, age 62.9 ± 7.54 years. The expression level of miRNA-155 and miRNA-146a was determined by real-time PCR. Results: The obtained data indicate an increase in the expression level of miRNA-146a and miRNA-155 in patients with clinical forms of endocrine ophthalmopathy at the stage of fibrosis (p <0.01). Conclusion: The obtained results of the functional activity of miRNA-146a and miRNA-155 confirm their participation in the pathogenesis of the development of edematous exophthalmos and endocrine myopathy by activating the fibroplastic process. Key words: endocrine ophthalmopathy, GC therapy, microRNA.

AB0114 INVESTIGATION OF THE EFFECTS OF EXERCISE ON MIRNA EXPRESSIONS IN PATIENTS WITH RHEUMATOID ARTHRITIS

Background:The goal of treatment for patients with RA is achieve to remission, or at least a state of low disease activity. Exercise is recommended for patients with RA in addition to drug therapy. It has been found to be effective in greatly improving functionality and reducing cardiovascular risk without exacerbating disease activity. Therefore, it is recommended that all RA patients should be encouraged to include aerobic and resistant exercise training as part of their routine treatment (1).miRNAs(miRNA) are known to protect the pathophysiological process specific to RA. miRNA-146a is one of the miRNAs extensively studied in RA, its expression was found to be higher in the synovial fluid and synovial tissue of RA patients compared to healthy individuals (2).Many studies have found that miRNA-146a, along with miRNA-16 and miRNA155 may be related to disease pathology. It has also been found that high levels of miRNA-16 expression correlate with active disease and low levels of expression with inactive disease. It has been found that the increased level of miRNA-155 causes a problem in the modulation of arthritis It has been found that the expression level of miRNA-145 is increased in peripheral blood mononuclear cells of RA patients and synovium supporting osteoclastogenesis (3,4,5).Objectives:It is aimed to investigate the effect of exercise on microRNA expressions in patients with rheumatoid arthritis (RA).Methods:30 patients and 30 healthy controls aged 18-60 years who met the 2010 ACR / EULAR RA criteria were included in the study. A program consisting of strengthening and stretching exercises 2 days a week was applied to the study group for 8 weeks. One day a week, 30 minutes of mild moderate walking was requested. Of the cases at the beginning and at the end of the treatment; 5-10 cc peripheral blood samples were taken into one EDTA tube. Then Numeric Rating Scale (NRS) was used for pain, 28-joint Disease Activity Score (DAS28) was used to calculate disease activity, Health Assessment Questionnaire (HAQ) was used to assess general health and Short Form-36 (SF-36) was used to evaluate quality of life. 5-10 cc peripheral blood samples were taken to only 1 EDTA tube of the control group. In the samples taken, gene expressions of miRNA-146a, miRNA-155, miRNA-16, miRNA-145 were determined by real-time PZR method.Results:There was a significant difference in DAS28, SF-36, NRS, HAQ scales before and after treatment in the RA group of patients (p 0.05). The expression level of MiRNA-146a does not differ significantly before and after treatment (p> 0.05). However, these two groups differ significantly with the control group (p 0.05). No significant difference was observed in the miRNA-155 and miRNA-16 expression levels in the pretreatment, posttreatment, and control groups (p> 0.05).Conclusion:Exercise therapy has a good effect on pain, disease activity, quality of life and general health in patients with RA. It has been found that exercise can affect vii some of the miRNAs involved in disease pathogenesis. However, more comprehensive studies are needed.

МикроРНК-146а – предиктор развития метастазов меланомы хориоидеи

MicroRNAs (miRNA, miR) - regulate gene expression, take part in the regulation of cell life, proliferation and apoptosis. There are not enough publications on the study of microRNA in the plasma of patients with choroidal melanoma (CM) and the presented data are contradictory. An increase in the level of microRNA-146a in blood plasma was noted in patients with cancer of the lung, esophagus and breast. In the present study, we used microRNA-146a in the blood plasma of patients with CM. Purpose. To determine the possibility and reliability of detecting microRNA-146a in the blood plasma of patients with CM, taking into account the localization of the tumor and its size. Material and methods. The blood plasma of 84 patients was studied mean age 63.4±1.2 years. A control group of 28 people who did not have tumor or chronic autoimmune diseases mean age 63.25±1.43 years. The expression level of miRNA-146a was determined by quantitative PCR. Results. An increase in the level of microRNA-146a was shown, which correlates with the localization and thickness of the tumor. The results were confirmed by statistical analysis. Conclusion. MicroRNA-146a, which increases in plasma levels in patients with CM as its size increases, can be used as a biomarker of aggressive tumor course in terms of hematogenous metastasis. Key words: choroidal melanoma, microRNA-146a, biomarkers.

The Expression of miRNA-146a in Peripheral Blood Mononuclear Cells of Patients with Myasthenia Gravis and Its Bioinformatics Analysis

The study is to investigate the expression of miRNA-146a in PBMC of myasthenia Gravis (MG), and to explore the molecular regulatory network of miRNA-146a in the pathogenesis of MG by bioinformatics. 108 patients with MG were selected as the experimental group (MG group), and 50 healthy subjects were selected as the control group. The relative expression of miRNA-146a in PBMC was detected by RT-PCR. The cross-target gene of miRNA-146a was predicted by TargetScan and CoMeTa database. miRNA-146a target gene GO enrichment and KEGG Pathway enrichment analysis was performed using the DAVID database. Our results shows that the expression level of miRNA-146a in peripheral blood of MG patients was significantly higher than that of the control group, the difference was statistically significant (P < 0 05), and the nucleotide sequence was highly conserved. The potential target genes of miRNA-146a include 88 kinds; GO analysis showed that miRNA-146a target gene function is mainly enriched in cell proliferation regulation, neuronal differentiation, etc. KEGG Pathway analysis shows that miRNA-146a is mainly enriched in Toll-like receptor signaling pathway, neurotransmitter regulatory signaling pathways, EB signaling pathways and other signaling pathways. In conclusion, the expression of miRNA-146a in PBMC of MG patients is up-regulated and participates in the pathogenesis of MG by acting on multiple signaling pathways.

A miRNA-Based Blood and Mucosal Approach for Detecting and Monitoring Celiac Disease.

The role of microRNAs (miRNAs) in celiac disease (CD) is unclear. We evaluated inflammation-related miRNA-146a, miRNA-155, miRNA-21, and miRNA-125b expression in peripheral blood and intestinal mucosa of CD adults. Thirty patients with CD were included: patients with active CD on a gluten-containing diet (CD-active, n = 10), patients on a gluten-free diet (for at least 1year), and patients with negative blood antibodies (CD-inactivePE, n = 10). In addition, ten healthy volunteers formed the comparison/control group. MiRNA expression was measured in duodenal biopsies from patients (CD-inactiveMU, n = 10) after in vitro exposure to PT gliadin and 33-mer peptide. MiRNAs expression was measured in plasma and in peripheral blood mononuclear cells (PBMCs) and monocytes, before and after in vitro exposure to native gliadin (gliadinN). Expression levels of miRNA-146a, miRNA-155, and miRNA-21 in PBMCs, miRNA-155 in monocytes and miRNA-155, miRNA-21, and miRNA-125b in plasma were elevated in both groups of celiac patients. After in vitro exposure with gliadinN, miRNA-146a and miRNA-155 expression markedly increased in PBMCs and monocytes, while miRNA-155 and miRNA-21 increased in the CD-active group. MiRNAs expression in intestinal mucosa did not change. MiRNA-146a and miRNA-155 expression showed high sensitivity and specificity for the presence of CD, irrespective of the current dietary treatment. Selected inflammation-related miRNAs expression is elevated in the peripheral blood of celiac. This suggests their participation in the immune processes underlying the pathology. Their similar response in active and inactive CD suggests that they should be further evaluated, as potential diagnostic biomarkers for CD.

Micro RNA in Degenerative Disorders

Obesity and type 2 diabetes mellitus considered as the most risk criteria in metabolic syndrome. Twenty four male albino rats, 3 months age weighting (120±15) gm at the beginning of the experiment, were purchased from animal house in our University. The rats were kept at a suitable conditions with a temperature of 23-25°C with 12 hours light-dark cycle and free availability of water and diet. The rats were kept for 2 weeks for accommodation, then randomly divided into 2 main groups the first one was a control group (8 rats) kept on a standard diet with free access to drinking water. The second group (16 rats) kept on high fat high fructose diet (HFHF), 8 rats kept for 7 weeks (considered as obese group G2) and the other 8 rats were kept for 12 week to be diabetic with type II according to (De Castro et al., 2013)and considered as diabetic group (G 3). Grossly, feeding HFHF diet resulted in marked increase in body weight in both obese and diabetic group, this increase was stopped between 7th and 8th week the body weight started to decrease. Laboratory, feeding HFHF diet resulted in decreasing the expression levels of insulin hormone gene which reflected on the increase in the expression levels of FGF-21, IL-1β, IL-6 and TNF-α genes. This increase was more in the diabetic group in IL-1β, and the same in both groups in IL-6 and TNF-α but higher in obese group in FGF-21 gene. The expression levels of miRNA 146a, miRNA 126, FGF 21 and some cytokines may be a good early indicators for obesity and diabetes mellitus type II, having a major role in the mechanism of incidence and may have a great role in the treatment and follow-up the disease.

Downregulation of miR-146a promotes cell migration in Helicobacter pylori-negative gastric cancer.

microRNAs (miRs) are short noncoding RNAs that post-transcriptionally suppress gene expression. miR-146a acts as an oncogene or a tumor suppressor in various cancers, including gastric cancer, but it is unclear what determines whether miR-146a is oncogenic or tumor suppressive and the molecular mechanisms are still largely unknown. The aim of this study was to investigate the role of miR-146a in gastric cancer, by focusing on its expression in patients who were negative for Helicobacter pylori and its reduced and increased expression effect in vitro. Twenty gastric cancer patients who were negative for H. pylori infection were selected and the expression levels of miRNA-146a in these gastric tumors, in their matched normal gastric tissues and in gastric cancer cell lines with varying tumorigenic potential was measured. Further, the impact of increased and decreased miR-146a expression levels on the expression of predicted target genes, cell migration, viability, proliferation, and apoptosis was examined, respectively. Our results for the first time indicated that miR-146a is downregulated in H. pylori-negative gastric cancers and suggests that H. pylori infection determines whether miR-146a acts as an oncogene or tumor suppressor. The level of miR-146a expression inversely correlates with the tumorigenicity of three gastric cancer cell lines and low miR-146a expression predicts poor recurrence-free survival. It was also found that miR-146a reduces the expression levels of the prometastatic genes and suppresses MKN-45 cell migration. Functional studies showed that miR-146a acts as a tumor suppressor miR and identifies miR-146a as a candidate for antimetastatic miRNA replacement therapy for gastric cancer patients.

Differential expression of miRNA-146a and miRNA-155 in gastritis induced by Helicobacter pylori infection in paediatric patients, adults, and an animal model

BackgroundHelicobacter pylori is a major aetiologic agent associated with gastritis. H. pylori infections increase the expression of the Toll-like receptor (TLR), which in turn modulates the expression of microRNA (miRNA)-146a and miRNA-155. The objective of this study was to compare the expression of miRNA-146a and miRNA-155 in gastric lesions of paediatric and adult patients with different pathologies and in Mongolian gerbils (Meriones unguiculatus) infected with H. pylori 26,695.MethodsQuantification of miRNA expression was performed by quantitative real-time polymerase chain reaction (qRT-PCR) of paraffin-embedded gastric lesions of children with or without an infection (n = 25), adults with follicular gastritis and metaplasia (n = 32) and eight-week-old M. unguiculatus males (Hsd:MON) infected with H. pylori 26,695 for 0, 3, 6, 12 and 18 months (n = 25). The genes RNU48 and RNU6 were used as endogenous controls for data normalization. Statistical analyses were performed using Kruskal-Wallis, Mann-Whitney, ANOVA and Student’s t-test.ResultsThe expression of miRNA-146a and miRNA-155 in infected children increased by 247.6- and 79.4-fold (on average), respectively, compared to that observed in the control group. However, these results were not significant (p = 0.12 and p = 0.07 respectively). In some children a gradual increase in expression was observed, while in others, expression was very high. Additionally, the expression levels of miRNA-146a and miRNA-155 increased by an average of 21.7- and 62-fold, respectively, in adult patients with follicular gastritis when compared to those of the controls. In M. unguiculatus infected with H. pylori 26,695, the expression of both miRNAs increased as the infection progressed.ConclusionThis is the first report to show differences in the expression of miRNA-146a and miRNA-155 in paediatric and adult patients with gastritis who were infected with H. pylori. In addition, in M. unguiculatus infected with H. pylori, miRNA expression was associated with the progression of infection and the ability of the bacteria to adapt to the host.

Evaluation of the miRNA-146a and miRNA-155 Expression Levels in Patients with Oral Lichen Planus.

Oral Lichen Planus (OLP) is a chronic autoimmune disease that could be considered as a potential premalignant status. To evaluate the miRNA-146a and miRNA-155 expression levels in patients with oral Lichen planus lesions compared to healthy subjects with normal oral mucosa. Forty patients with oral lichen planus and 18 healthy age and gender-matched controls were recruited in this case-control study. Oral lichen planus was diagnosed clinically and pathologically. The expression levels of two miRNAs in peripheral blood samples were determined using commercial TaqMan MicroRNA Assays. Relative quantification of gene expression was calculated by the 2-ΔΔct method. The expression levels of miRNA-146a and miRNA-155 in patients with oral Lichen planus were significantly higher than those of healthy controls. Also, a direct but insignificant correlation was found between miRNA-155 and miRNA-146a expression levels among the patient group. Our findings indicate that miRNA-146a and miRNA-155 could be potential biomarkers for the immunopathogenesis of oral lichen planus.

MiRNA-146a, miRNA-155 and JNK expression levels in peripheral blood mononuclear cells according to grade of knee osteoarthritis

Osteoarthritis (OA) is the most common joint disease characterized by joint pain and a progressive loss of articular cartilage. OA known as a non-inflammatory disease. Despite this the recent studies are shown synovitis and low inflammation to have a role in OA pathophysiology. The aim of this study to determine the roles of a potential therapeutic targets miRNA-146a, miRNA-155 and JNK expression levels in OA patients. Peripheral mononuclear blood cells (PBMCs) were extracted from OA patients and healthy subjects. The expression levels of miRNA-146a, miRNA-155 and JNK were quantified using by real-time PCR assay. According to study results a statistically significant increase was observed only in miRNA-155 expression level (p=0,039). However, miRNA-146a and miRNA-155 expressions increased in the progressive stages (grade 3 and grade 4) in OA patients. Our data suggests that correlation of miRNAs regulating and signal pathways can play an important role in OA pathogenesis and disease progression.

Expression of miRNA-155 and miRNA-146a in peripheral blood mononuclear cells and plasma of oral lichen planus patients

To investigate the expression and clinical significance of miRNA-155 and miRNA-146a in peripheral blood mononuclear cells (PBMC) and plasma of oral lichen planus (OLP) patients. Twenty-five female and seven male OLP patients (OLP group) aged 25 to 54 years were selected from January 2012 to May 2013. The diagnosis was confirmed by pathology and the lesions were divided into two non-erosive OLP group (18 cases) and erosive OLP group (14 cases). Twenty healthy sex and age matched volunteers served as control. miRNA-155 and miRNA-146a expressions in PBMC and plasma were examined by real-time PCR. The difference between OLP group and control group was statistically analyzed. The expressions of PBMC and plasma miRNA-155 were higher in OLP patients than those in the healthy control (median, 0.07 vs 0.03, P < 0.05; 5.84 vs 1.32, P < 0.01). The median expression level of miRNA-146a in PBMC and plasma of OLP patients and healthy controls were (1.26 vs 0.58, P < 0.05) and (412.60 vs 238.42, P < 0.01). The plasma miRNA-155 and miRNA-146a expressions were significantly higher in erosive OLP group than those in non-erosive OLP group. There were no significant differences in the expression of PBMC miRNA-155 and miRNA-146a between the two groups. The expressions of PBMC and plasma miRNA-155 and miRNA-146a are higher in OLP patients. The expressions of plasma miRNA-155 and miRNA-146a are associated with OLP severity. The over expression of miRNA-155 and miRNA-146a in OLP may play a role in the pathogenesis of OLP.

Expression of microRNA146a in ultraviolet B (UVB)-damaged mouse skin

Objective To investigate the expression changes of microRNA 146a (miRNA146a) in UVBdamaged mouse skin. Methods C57/BL6 mice were divided into dose groups to be irradiated on the back with UVB of 30, 60, 90, 180, 270 mJ/cm2 respectively for 24 hours, and time groups irradiated with UVB of 180 mJ/cm2 for 1, 12, 24 and 48 hours respectively. The mice received no irradiation served as the control. Skin samples were subsequently obtained from the irradiated sites of mice and subjected to real-time fluorescent PCR for the detection of miRNA146a expression as well as immunohistochemical staining (IHC) for the detection of STAT3. Results The expression levels of miRNA146a were 0.01158 ± 0.00098, 0.01083 ± 0.00104,0.00872 ± 0.00031, 0.00851 ± 0.00033, 0.00810 ± 0.00057 and 0.00770 ± 0.00031 in the unirradiated control mice and mice irradiated with UVB of 30, 60, 90, 180, 270 mJ/cm2 for 24 hours, respectively, 0.00730 ±0.00036, 0.00805 ± 0.00035, 0.00810 ± 0.00057 and 0.00837 ± 0.00039 in mice irradiated with UVB of 180mJ/cm2 for 1, 12, 24 and 48 hours, respectively. IHC suggested an intensive expression of phosphorylated STAT3 in mice irradiated with a high dose UVB. Conclusions miRNA146a may play an essential role in the mechanism of UVB-induced photodamge, which is mainly correlated with the negative regulation of inflammatory reaction likely mediated by the JAK/STAT3 signal transduction pathway. Key words: MicroRNAs; Radiation injuries, experimental; STAT3 transcription factor