The ABO blood group antigens in patients with oral lichenoid reaction.

Evaluating the association of ABO blood group with different delayed hypersensitivity reactions, such as oral lichenoid reaction (OLR), can provide a new perspective for clinical practice. Therefore, this study designed to investigate ABO blood group antigens in OLR patients. In this case-control study, the ABO blood group of 112 OLR patients and 117 individuals without oral lesions were included. Gender, age, characteristics of the lesions, medications and restorative materials recorded. Chi-square test used to compare the frequency of ABO blood groups in OLR patients with controls. The O blood group was significantly higher in OLR patients and all its subtypes. Also, there were significant relation between O blood group, and severity of lesions. The frequency of dysplasia was non-statistically significant higher in OLR patients with O blood group than other blood group. Based on the results of the present study, O blood group was significantly more in patients with lichenoid reaction than control group, and AB blood group was the lowest. Also, O blood group showed a positive association with the more severe form of OLR lesions and frequency of dysplasia.

Unveiling the Potential of Serum MiR-483-5p: A Promising Diagnostic and Prognostic Biomarker in OLP and OSCC Patients by In silico Analysis of Differential Gene Expression.

BACKGROUND: Oral squamous cell carcinoma (OSCC) and oral lichen planus (OLP) are two separate conditions affecting the mouth and result in varying clinical outcomes and levels of malignancy. Achieving early diagnosis and effective therapy planning requires the identification of reliable diagnostic biomarkers for these disorders. MicroRNAs (miRNAs) have recently received attention as powerful biomarkers for various illnesses, including cancer. In particular, miR-483-5p is a promising diagnostic and prognostic biomarker in various cancers. Therefore, this study aimed to investigate the role of serum miR-483-5p in the diagnosis and prognosis of OLP and OSCC patients by in silico analysis of differential gene expression. METHODS: GSE23558 and GSE52130 data sets were selected, and differential gene expression analysis was performed using microarray data from GSE52130 and GSE23558. The analysis focused on comparing OLP and OSCC samples with normal samples. The genes intersected through the differential gene expression analysis were then extracted to determine the overlapping genes among the upregulated or downregulated DEGs. The downregulated genes among the DEGs were subsequently imported into the miRWalk database to search for potential target genes of miRNA 483-5p that lacked validation. To gain insight into the biological pathways associated with the DEGs, we conducted pathway analysis utilizing tools, such as Enrichr. Additionally, the cellular components associated with these DEGs were investigated by analyzing the String database. On the other hand, blood serum samples were collected from 35 OSCC patients, 34 OLP patients, and 34 healthy volunteers. The expression level of miR-483-5p was determined using quantitative reverse transcription polymerase chain reaction (RT-qPCR). The Kruskal-Wallis test was utilized to investigate the considerable correlation. Moreover, this study explored the prognostic value of miR-483-5p through its association with clinicopathological parameters in OSCC patients. RESULTS: The results showed that serum expression of miR-483-5p was considerably higher in OSCC patients compared to OLP patients and healthy controls (p 0.0001) and that this difference was statistically significant. Furthermore, elevated miR-483-5p expression was associated with tumor size, lymph node metastasis, and stage of tumor nodal metastasis in OSCC patients (p 0.001, p 0.038, and p 0.0001, respectively). In silico analysis found 71 upregulated genes at the intersection of upregulated DEGs and 44 downregulated genes at the intersection of downregulated DEGs, offering insight into the potential underlying mechanisms of miR-483-5p's engagement in OSCC and OLP. The majority of these DEGs were found to be involved in autophagy pathways, but DEGs involved in the histidine metabolism pathway showed significant results. Most of these DEGs were located in the extracellular region. After screening for downregulated genes that were invalidated, miRNA 483-5p had 7 target genes. CONCLUSION: This study demonstrates the potential of serum miR-483-5p as a promising diagnostic and prognostic biomarker in OSCC and OLP patients. Its upregulation in OSCC patients and its association with advanced tumor stage and potential metastasis suggest the involvement of miR-483-5p in critical signaling pathways involved in cell proliferation, apoptosis, and cell cycle regulation, making it a reliable indicator of disease progression. Nevertheless, additional experimental studies are essential to validate these findings and establish a foundation for the advancement of targeted therapies and personalized treatment approaches.

Follicular-Stimulating Hormone, Luteinizing Hormone, and Prolactin Serum Level in Patients with Oral Lichen Planus in Comparison to Healthy Population.

MATERIALS AND METHODS: In this cross-sectional case control study, the serum level of LH, FSH, and prolactin of 40 women with lichen planus who have been referred to Shiraz Dental Faculty, Oral and Maxillofacial Disease Department during 2018-2019 has been evaluated in comparison to 40 healthy controls. Data were analyzed by SPSS version 18. Two-way ANOVA and Mann-Whitney test were used for data analysis. RESULTS: The mean serum level of FSH and LH was significantly higher in OLP patients while this difference was not reported for prolactin. Only FSH mean serum level was significantly higher in nonmenopausal OLP patients. The distribution of prolactin and FSH hormones' serum level was in normal range. CONCLUSIONS: The high serum level of FSH and LH can affect OLP pathogenesis by estrogen and progesterone modulation.

Systematic Review and Meta-Analysis of Oxidative Stress and Antioxidant Markers in Oral Lichen Planus.

BACKGROUND: Oral lichen planus (OLP) is a relatively common chronic inflammatory disease of unknown etiology, which might be caused by oxidative stress and impaired antioxidant defense. OBJECTIVE: To systematically investigate the markers of oxidative stress and antioxidant systems in the saliva and blood from OLP patients and healthy controls. METHODS: The PubMed, Cochrane Library, and Embase were systematically queried to collect data from studies in which oxidative stress/antioxidant markers from OLP and healthy subjects had been evaluated until March 10, 2021. RESULTS: A total of 28 studies fulfilled inclusion criteria, and 25 of them, having 849 OLP patients and 1,052 control subjects and analyzing 12 oxidative stress and antioxidant state marker levels, were subjected to meta-analysis. We found a significant decrease in total antioxidant capacity (TAC) and uric acid (UA) and a significant increase in malondialdehyde (MDA) and nitric oxide (NO) levels in the saliva and serum/plasma of OLP patients. Moreover, a significant elevation of 8-hydroxy-deoxyguanosine (8-OHdG) and advanced oxidation protein product (AOOP) level and a decrease in vitamin C were also observed in the saliva of the OLP group. In contrast, salivary vitamin A, zinc, glutathione peroxidase (GPx), vitamin E, and nitrite were not significantly different between the two groups. In single studies, markers of oxidative stresses such as superoxide dismutase (SOD) and 8-isoprostanelevels were elevated in OLP, and antioxidant parameters such as glutathione (GSH) and total protein (TP) levels were dysregulated. CONCLUSION: This meta-analysis helps to clarify the profile of oxidative stress and antioxidant state markers in OLP patients although existing evidence is rather heterogeneous and many studies are affected by several limitations. Larger and more standardized studies are warranted to ascertain whether these markers are potential causes or effects of OLP and whether antioxidant therapy improving oxidative stress will be useful.

Evaluation of correlation between transcription factors and IL-17 in oral and cutaneous lichen planus lesions and Leukocytes.

BACKGROUND: Lichen planus (LP) is a chronic autoimmune disease with different clinical subtypes including cutaneous LP (CLP) and oral LP (OLP). We aimed to compare mRNA expression of RORγt and IL-17 in paraffin-embedded blocks of OLP and CLP lesions with normal oral mucosa (NOM), and also its correlation with hematologic parameters. MATERIALS & METHODS: This study included 89 paraffin-embedded blocks contain OLP (44 cases), CLP (45 cases) and NOM from the archive of Mashhad University of Medical Sciences, Mashhad, Iran. The expression of RORγt and IL-17 was evaluated by Real-time RT-PCR method. The result was compared to Leukocyte counts and the other hematological parameters of studied patients. RESULTS: The results of our study showed IL-17 and RORγt expression in OLP lesions were significantly higher than CLP and NOM groups (P = 0.001). Although we found high expression of RORγt and IL-17 in erosive OLP in compared to classic OLP lesion, but this increment was not significant for IL-17 (P = 0.26) and RORγt (P = 0.14). Further, Leukocyte and monocyte counts were substantially high in OLP group in compared to the CLP and NOM groups (P < 0.05). CONCLUSIONS: We concluded that increased expression of RORγt and IL-17 in LP lesions could play role in the pathogenesis of LP. As well, higher expression of RORγt and IL-17 in oral LP more than cutaneous LP might be associated with difference in clinical behavior of the two types of disease and role of these factors in premalignant behavior of OLP lesions.

New markers of oxidative stress in lichen planus and the influence of hepatitis C virus infection - a pilot study.

Introduction. Lichen planus (LP) is a mucocutaneous T-cell mediated disorder of unknown etiology. There is growing evidence that oxidative stress is an important player in the pathogenesis of LP. Therefore, we have investigated oxidative stress markers in LP and the influence of hepatitis C virus (HCV) infection, a frequently associated condition, on oxidative stress in LP patients. Method. We have determined the serum levels of 4- hydroxynonenal (4-HNE) and symmetric dimethylarginine (SDMA), as markers of oxidative stress, and total antioxidant capacity (TAC), as a marker of the antioxidant defence, in 4 groups: group A - HCV positive patients with LP (n=12), group B - HCV positive patients without LP (n=12), group C - HCV negative patients with LP (n=31) and group D - control group (n=26). Results. In LP patients, we have identified an increased level of lipid peroxidation (4-HNE - group A - 8.41±1.11 µg/mL, group B - 7.97±2.17 µg/mL, group C - 7.81±1.96 µg/mL and group D - 6.15±1.17 µg/mL) and alterations in arginine methylation (SDMA - group A - 1.10±0.24 µmol/L, group B - 1.03±0.16 µmol/L, group C - 0.84±0.19 µmol/L and group D - 0.50±0.06 µmol/L) associated with a diminished antioxidant defence (TAC - group A - 234.50±49.96, µmol/L group B - 255.83±41.41 µmol/L, group C - 269.83±43.33 µmol/L and group D - 316.46 ±29.33 µmol/L), processes augmented by the association with HCV infection. Conclusion. There is an imbalance between oxidants and antioxidants in patients with LP, an imbalance that is augmented by the presence of HCV infection. SDMA could be regarded as a novel biomarker of oxidative stress among these patients. To the best of our knowledge this is the first study to investigate the influence of HCV infection on oxidative stress in LP patients.

Serum Vitamin Profile in Oral Lichen Planus Patients in Southwest of Iran.

INTRODUCTION: Oral lichen planus (OLP) is a chronic mucocutaneous disease. It is mainly an immune system-related disorder. Vitamins can modulate immune system functions, and thus, vitamin deficiency might have roles in exacerbating OLP. We aim to determine the serum levels of vitamins A, B12, C, D3, and E in OLP patients. METHODS AND MATERIALS: 34 OLP patients referred to Shiraz Dental School entered the study. Blood samples were collected and levels of A, B12, C, D3, and E vitamins were measured in serum. 43 healthy people were also included as the control group. Serum levels of vitamins were measured by HPLC (A, B12, D3, and E) and Kiazist analyzing kit (vitamin C). RESULTS: Most of the patients were female (62.3%), and the mean age of patients was 48.03 ± 11.57. Serum levels of vitamins A, C, and E were lower in OLP patients in comparison with the healthy group; however, the difference was not significant. Vitamins B12 and D3 were higher in the OLP group but the difference was not significant. CONCLUSION: Serum levels of vitamins A, B12, C, D3, and E do not have a significant difference in OLP patients and healthy groups. These vitamins may not have a considerable role in OLP pathogenesis in the southwest of Iran.

Serological and psychological assessment of patients with oral lichen planus using serum cortisol levels and hads questionnaire-a case control study.

BACKGROUND: Oral lichen planus (OLP) is a T cell-mediated chronic autoimmune disorder directed against antigens secreted by the basal cell layer, with an incidence of 0.02-0.22% in Indian population and showing female predilection. Stress is considered one of the etiological factors in the causation, progression, and recurrence of this disease. OBJECTIVES: To evaluate the levels of serum cortisol, anxiety, and depression in patients with symptomatic OLP and to correlate the levels of serum cortisol with anxiety and depression. METHODS: Sixty subjects were categorized into two groups. Group A: 30 adults with no history of OLP and no psychological history of anxiety and depression. Group B: 30 patients with clinically and histopathologically diagnosed symptomatic OLP. The subjects in both groups were evaluated for anxiety and depression levels using the Hospital Anxiety and Depression Scale (HADS) questionnaire and serum cortisol levels (8-9 am sample) using the chemiluminiscence method. RESULTS: Higher depression and anxiety levels were significantly associated with group B with significant P values (P < 0.0001 and <0.0002 respectively) when compared with group A; higher mean serum cortisol levels were seen in group B compared with group A, with P < 0.0001. In group A, a positive correlation was found between depression, anxiety, and serum cortisol levels with non-significant P-value. In group B, a positive correlation was found between depression, anxiety, and serum cortisol levels with a significant P value (P < 0.0001). CONCLUSION: Increased levels of depression and anxiety with increased serum cortisol levels were observed in subjects with OLP.

[Effects of interleukin-35 on the balance of helper T cell 17/regulatory T cell in peripheral blood of patients with oral lichen planus].

Objective: To investigate the effects of exogenous interleukin (IL)-35 on the balance of helper T cell 17 (Th17) and regulatory T cell (Treg) in peripheral blood of patients with oral lichen planus (OLP). Methods: Totally 12 peripheral blood samples of OLP patients (OLP group, one male and 11 female, 26-68 years old; four cases of reticular OLP and eight cases of erosive OLP) were collected from patients of Department of Oral Mucosal Specialist of the Affiliated Hospital of Guizhou Medical University from October to December 2016. During the same period, thirteen normal peripheral blood samples were collected from the Physical Examination Center of the Affiliated Hospital of Guizhou Medical University (normal control group, one male and 12 female, 20-68 years old). The peripheral blood mononuclear cells (PBMC) were extracted in sterile condition and CD4+ T cells were sorted by flow cytometry (FCM). Quantitative real-time PCR (qPCR) technique was used to detect the mRNA expression levels of retinoid-related orphan nuclear γt (RORγt) and forkhead box3 (Foxp3). The CD4+ T cells were divided into experimental group and control group. The CD4+ T cells of experimental group were cultured in vitro by adding rhIL-35, and the CD4+ T cells of control group were cultured with the same volume of phosphate buffered saline (PBS). After the completion of the culture, the cells were collected. The expression levels of the same factors were detected by qPCR. Results: The expression [M(Q(25), Q(75))] of Foxp3 [0.15 (0.09, 0.30)] and RORγt mRNA [1.04 (0.45, 2.15)] in the CD4+ T cells of OLP were significantly higher than those in normal control group [0.04 (0.02, 0.06), 0.10 (0.05, 0.11)] (Z=-4.134, P<0.01; Z=-3.699, P<0.01). The ratio of ROR γt/Foxp3 mRNA in OLP group [6.22(3.67, 15.34)] was higher than that in normal control group [2.50 (1.24, 5.23)] (Z=-2.665, P=0.007). In the CD4+ T cells of OLP patients, the expression of Foxp3 mRNA in the experiment group [0.40 (0.21, 1.22)] was higher than that in the control group [0.15 (0.11, 0.26)](Z=-2.510, P=0.012), and the expression of ROR γt mRNA between two groups showed no significant difference (P>0.05). The ROR γt/Foxp3 mRNA ratio [3.44 (1.55, 8.16)] of the experiment group was lower than that in the control group [6.22 (4.43, 12.21)] (Z=-2.746, P=0.006). Conclusions: There was a Th17/Treg imbalance with predominated by Th17 cells in the peripheral blood of patients with OLP. Exogenous rhIL-35 had an immunomodulatory effect on the balance of Th17/Treg.

Salivary and Serum Interferon-Gamma/Interleukin-4 Ratio in Oral Lichen Planus Patients: A Systematic Review and Meta-Analysis.

Background and Objectives: Interferon-gamma (IFN-γ)/interleukin-4 (IL-4) ratio may indicate a change in the immune response with a potential pathological effect presented in oral lichen planus (OLP) patients. Herein, this meta-analysis evaluated the role of serum and salivary interferon-gamma/interleukin-4 ratio in the severity and development of OLP. Materials and Methods: The Scopus, Cochrane Library, PubMed, and Web of Science databases were systematically searched to retrieve the relevant studies published up from the database inception to March 2019. The crude mean difference (MD) and 95% confidence interval (CI) were calculated by RevMan 5.3 software using a random-effects model. A sensitivity analysis was performed on the results using the CMA 2.0 software. A total of 98 studies were retrieved from the databases, of which at last seven studies were included in this meta-analysis. Results: The findings showed that the pooled MDs of serum and salivary IFN-γ/IL-4 ratio were -0.22 (95% CI: -1.16, 0.72; p = 0.64) and 0.17 (95% CI: -1.50, 1.84; p = 0.84) in OLP patients compared to controls, respectively. In addition, the pooled MDs of serum and salivary IFN-γ/IL-4 ratio were -0.15 (95% CI: -0.53, 0.23; p = 0.43) and -0.39 (95% CI: -0.63, -0.15; p = 0.001) in patients with erythematous/ulcerative subtype compared to patients with reticular subtype, respectively. Conclusions: In conclusion, the results of meta-analysis demonstrated that serum and salivary IFN-γ/IL-4 ratio cannot play a major role in OLP development and severity.

Expression and biological functions of the CCL5-CCR5 axis in oral lichen planus.

Oral lichen planus (OLP) is a chronic inflammatory oral mucosal disorder with T cell-mediated immunological pathogenesis. C-C motif chemokine ligand 5 (CCL5) and its receptor C-C motif chemokine receptor 5 (CCR5) play important roles in the activation and recruitment of T cells. This study sought to explore the expression and biological functions of the CCL5-CCR5 axis in OLP. We examined the expression of the CCL5-CCR5 axis in the peripheral blood and oral tissue of healthy controls and patients with OLP using quantitative real-time PCR, Simple Western assays, enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. In addition, we investigated the effects of the CCL5-CCR5 axis on the proliferation, apoptosis and migration of OLP T cells using Cell Counting Kit-8 (CCK8) assay, flow cytometry and transwell assay. We found that the expression of the CCL5-CCR5 axis was significantly elevated both in the peripheral blood and oral tissue of patients with OLP compared with healthy controls. CCL5 not only promoted OLP T-cell proliferation and migration but also inhibited OLP T-cell apoptosis. Moreover, CCR5 inhibition suppressed OLP T-cell proliferation and migration, whereas OLP T-cell apoptosis was promoted. In conclusion, our data suggest that the CCL5-CCR5 axis may be closely related to the inflammatory infiltration of T cells in OLP.

Serum-based metabolomics characterization of patients with reticular oral lichen planus.

OBJECTIVE: Oral lichen planus (OLP) is a chronic inflammatory mucosal lesion and systemic disease. In OLP, reticular type is the most common presentation of the disease. However, little is known about it. The aim of this study was to analyze the pathogenesis of reticular OLP and its possible associations with the pathological changes in other organ systems through serum-based metabolomics. METHODS: Blood samples were obtained from 16 reticular OLP patients and 24 control subjects. Liquid chromatography (LC)-mass spectrometry (MS) system was used to identify differentially expressed metabolites. The pathways analysis was performed by MetaboAnalyst. Pathological network was constructed by Cytoscape software. RESULTS: Totally, 31 modulated metabolites were identified, whose dysregulations affected 25 metabolic pathways and 7 pathological processes in the disease. Through an impact-value screen (impact-value＞0.1), 6 pathways were selected as the significantly dysregulated pathways. Pathological network showed that these metabolites participated in 7 pathological processes, that is, apoptosis process, DNA damage and repair disorder, oxidative stress injury, carbohydrate metabolism disorder, mood dysfunction, inflammatory lesion, and other pathological process. CONCLUSION: The study demonstrated that reticular OLP could cause the dysregulations of the metabolites in serum, which might be also further linked to other organ and systemic diseases through the blood system, such as diabetes, sleep disorders, and depression, etc.

Evaluation of the serum levels of galectin-3 in patients with oral lichen planus disease.

OBJECTIVES: Galectin-3, a member of beta-galactoside-binding proteins, can be found in cytoplasm and nucleus as well as extracellularly in various tissues and involved in many physiological and pathological processes. We aimed to measure the serum levels of galectin-3 in oral lichen planus (OLP) disease and compare the result with that observed in healthy ones. MATERIALS AND METHODS: In this cross-sectional study, the serum levels of galectin-3 were measured in 56 healthy individuals and 53 pathologically proven OLP patients including those with atrophic/erosive (33 cases) or reticular (20 cases) lesions, using enzyme-linked immunosorbent assay (ELISA). RESULTS: Compared with healthy individuals (1.1 ± 0.4 ng/ml), galectin-3 serum levels in patients with OLP (3.1 ± 1.1 ng/ml) were significantly elevated (p < 0.0001). Serum galectin-3 levels were elevated significantly in patients with atrophic/erosive lesions compared to those of reticular (3.9 ± 2.1 ng/ml vs. 1.9 ± 1.4 ng/ml, p = 0.001), but it has not associated with age and gender. CONCLUSIONS: The elevation of galectin-3 in OLP is a future tool to increase the knowledge about the possible etiology of the disease and may be helpful to differentiate atrophic/erosive lesions from reticular ones. To the best of our knowledge, this is the first study evaluated the serum galectin-3 levels in OLP.

Expression and significance of phosphodiesterase 4B gene in peripheral blood of patients with oral lichen planus.

BACKGROUND: The pathogenesis of oral lichen planus (OLP) may be related to mental factors. In this study, we investigated the molecular mechanism of mental factors in the development of OLP. METHODS: The normal control group and OLP patients were tested and evaluated by Zung self-rating anxiety scale and self-rating depression scale. Secondly, Agilent human LncRNA chip technology was used to detect differential genes in the total RNA of the normal control group and OLP patients, and to screen out the differentially expressed genes related to anxiety and depression. Thirdly, we verified the genes at gene level and protein level, respectively. RESULTS: Zung self-rating anxiety scale and self-rating depression scale showed that tendency of anxiety and depression in OLP patients is significantly higher than that in normal controls, but there was no significant difference between the erosion form group and the reticular form group; the duration of OLP showed significant negative correlations between degree of anxiety and depression. Microarray analysis showed there were four differential genes (PDE4B, RGS2, SYNGR1, and SYNE1) related to anxiety and depression in OLP patients; real-time qPCR confirmed the expression of PDE4B mRNA was lower in the peripheral blood of normal control group (P < 0.001). The expression of RGS2, SYNGR1, and SYNE1 mRNA was higher in the normal control group (P < 0.05, P < 0.05, P < 0.05). Wes automatic western blot confirmed that the expression of PDE4B protein was lower in the peripheral blood of the normal control group (P < 0.01). CONCLUSION: PDE4B gene may play a role in the pathogenesis of OLP.

Serum and Salivary IgA, IgG, and IgM Levels in Oral Lichen Planus: A Systematic Review and Meta-Analysis of Case-Control Studies.

Immunoglobulins (IgA, IgG, and IgM) are significant anti-inflammatory factors. The meta-analysis aimed to assess the serum and salivary levels of Igs as more important immunoglobulins in patients affected by oral lichen planus (OLP) compared to the healthy controls. Four databases, including PubMed/Medline, Scopus, Web of Science, and Cochrane Library as well as Iranian databases were checked up to January 2018 without language restriction. The quality of each involved study was done using the Newcastle⁻Ottawa Quality Assessment Scale (NOS) questionnaire. A random-effects model analysis was done by RevMan 5.3 software applying the mean difference (MD) plus 95% confidence intervals (CIs). The CMA 2.0 software was applied to calculate the publication bias among the studies. Out of 70 studies found in the databases, 8 studies were involved and analyzed in the meta-analysis. The meta-analysis included 282 OLP patients and 221 healthy controls. The pooled MDs of serum levels of IgA, IgG, and IgM were -0.13 g/L [95% CI: -0.24, -0.02; P = 0.02], 1.01 g/L [95% CI: -0.91, 2.93; P = 0.30], and -0.06 g/L [95% CI: -0.25, 0.14; P = 0.56], respectively; whereas, the salivary IgA and IgG levels were 71.54 mg/L [95% CI: 12.01, 131.07; P = 0.02] and 0.59 mg/L [95% CI: -0.20, 1.38; P = 0.14], respectively. Considering the few studies performed on saliva, the results suggested that the salivary levels, especially IgA level had higher values than the serum levels. Therefore, the salivary immunoglobulins can play a significant function in the OLP pathogenesis.

Forkhead box p3 controls progression of oral lichen planus by regulating microRNA-146a.

Oral lichen planus (OLP) is a severe T cell-mediated disorder of the mucosa, which causes chronic inflammation. Forkhead box P3 (Foxp3) regulates the immune response and plays an important role in immunological diseases. The current study aimed to determine the role of Foxp3 and microRNA (miR)-146a in OLP. Western blot analysis and a quantitative real-time polymerase chain reaction assay showed that the expression of Foxp3 and miR-146a was upregulated in OLP tissues and in lipopolysaccharide (LPS)-incubated HaCaT cells, compared with controls. Foxp3 inhibition significantly decreased miR-146a expression, ameliorated LPS stimulation by decreased cell proliferation, and apoptosis in LPS-incubated HaCaT cells as compared with the LPS group. Cotransfection of Foxp3 small interfering RNA and miR-146a mimics elevated cell proliferation and apoptosis compared with the Foxp3 small interfering RNA group. In addition, miR-146a overexpression upregulated, whereas miR-146a inhibition downregulated, the proliferation and apoptosis of LPS-incubated HaCaT cells. The target gene of miR--146a, tumor necrosis factor receptor-associated factor 6 (TRAF6), was predicted by bioinformatics software and identified by the luciferase reporter assay. Furthermore, Foxp3/miR-146a elevated T regulatory cells and regulated TRAF6 expression in CD4+ T cells that were isolated from peripheral blood of patients with OLP. In conclusion, our study suggests that Foxp3 and miR-146a regulate the progression of OLP by negatively regulating TRAF6, which may provide a promising therapeutic target for OLP treatment.

Comparison of Plasma Exosomes by Differential Ultracentrifugation and Solvent Precipitation Methods.

BACKGROUND: Emerging evidence has identified that exosomes play a pivotal role in intercellular signal transmission. However, the standardized purification techniques to isolate high quality exosomes are still deficient at present. This study was to evaluate reproducibility and efficiency of differential ultracentrifugation and solvent precipitation-based kits by isolating plasma-derived exosomes from oral lichen planus patients. METHODS: Morphology, exosomal biomarkers, particle size distribution, proteomic components, and protein yield of isolated exosomes were evaluated by transmission electron microscope, western blot, laser diffraction instrument, Coomassie staining, and BCA protein assay kit, respectively. RESULTS: TEM displayed representative cup-shaped morphology of exosomes and western blot identified exosomal biomarkers CD9 and CD63. The size distribution showed that particles by differential ultracentrifugation were mainly from 26.15 nm to 166.5 nm, while some of the particles obtained by solvent precipitation kits were larger than 1,000 nm. In addition, exosomes isolated by solvent precipitation kits showed a significantly higher amount of protein yield due to plasma albumin contamination. CONCLUSIONS: Both differential ultracentrifugation and precipitation based kits could successfully isolate plasma exosomes, and exosomes by differential ultracentrifugation were purer and more appropriate for further proteomic analysis.

Gastric parietal cell and thyroid autoantibodies in recurrent aphthous stomatitis patients with concomitant oral lichen planus.

BACKGROUND/PURPOSE: Gastric parietal cell antibody (GPCA), thyroglobulin antibody (TGA), and thyroid microsomal antibody (TMA) have not yet been reported in recurrent aphthous stomatitis (RAS) patients with concomitant oral lichen planus (OLP/RAS patients). This study mainly assessed the frequencies of serum GPCA, TGA, and TMA (GPCA/TGA/TMA) positivities in 44 OLP/RAS patients. METHODS: The frequencies of serum GPCA/TGA/TMA positivities in 44 OLP/RAS patients, OLP/RAS patients of four different subgroups, 520 RAS patients, and 352 healthy control subjects were calculated and compared. RESULTS: We found that 20.5%, 27.3%, and 31.8% of 44 OLP/RAS patients, 75.0%, 100.0%, and 100.0% of 4 OLP/major-typed RAS (OLP/major RAS) patients, 15.0%, 20.0%, and 25.0% of 40 OLP/minor-typed RAS (OLP/minor RAS) patients, 45.5%, 72.7%, and 54.5% of 11 atrophic glossitis-positive OLP/RAS (AG+OLP/RAS) patients, and 12.1%, 12.1%, and 24.2% of 33 AG-negative OLP/RAS (AGÖ¾OLP/RAS) patients had the presence of GPCA, TGA, and TMA in their sera, respectively. OLP/RAS patients and OLP/RAS patients of four different subgroups all had significantly higher frequencies of GPCA/TGA/TMA positivities than healthy control subjects. Moreover, OLP/RAS patients had a significantly higher frequency of TMA positivity than RAS patients, and OLP/major RAS and AG+OLP/RAS patients had significantly higher frequencies of GPCA/TGA/TMA positivities than RAS patients. Furthermore, OLP/major RAS patients had significantly higher frequencies of GPCA/TGA/TMA positivities than OLP/minor RAS patients. CONCLUSION: For OLP/RAS patients, the concomitant OLP may play a role in causing an increased frequency of TMA positivity, and major RAS and the concomitant AG are contributory factors causing the elevated frequencies of GPCA/TGA/TMA positivities.

MicroRNA Microarray-Based Identification of Involvement of miR-155 and miR-19a in Development of Oral Lichen Planus (OLP) by Modulating Th1/Th2 Balance via Targeting eNOS and Toll-Like Receptor 2 (TLR2).

BACKGROUND A wide range of microRNAs (miRNAs) have been shown to play a significant role in disease regulation. The objective of this study was to explore the role of miR-155 and miR-19a in the regulation of oral lichen planus (OLP). MATERIAL AND METHODS Microarray assay, real-time PCR, Western blot assay, computational analysis, luciferase assay, ELISA, and immunohistochemistry analysis were carried out to investigate the role of miR-155 and miR-19a in OLP. RESULTS According to microarray assay and real-time PCR results, the expression of miR-155 was most significantly decreased among the 16 candidate miRNAs in the OLP group, whereas the expression of miR-19a was most significantly increased. MiR-155 and miR-19a directly targeted endothelial nitric oxide synthase (eNOS) and TLR2, respectively, since only the cells co-transfected with miR-155/wild-type eNOS 3'UTR or cells co-transfected with miR-19a/wild-type TLR2 3'UTR exhibited decreased luciferase activity. In addition, the expression of TLR2 was highly upregulated in OLP, whereas the expression of eNOS was significantly downregulated. A negative correlation was found between miR-19a and TLR2 mRNA, with a coefficient value of -0.40. Similarly, a negative correlation was found between miR-155 and eNOS mRNA, with a coefficient value of -0.54. A lower level of NO, IL-4, IL-5, and IL-10 was observed in OLP, which was also accompanied by a higher level of TNF-α and IFN-γ. Finally, the upregulation in miR-155 directly decreased the expression of eNOS and further inhibited the production of NO. Downregulation of miR-19a directly increased the expression of TLR2. The inhibition of NO production and the enhancement in TLR2 expression synergistically increased the production of TNF-α and IFN-γ, while decreasing the levels of IL-4, IL-5, and IL-10. CONCLUSIONS In this study, the peripheral blood mononuclear cells (PBMCs) from subjects with or without OLP were collected and their gene expression profiles were compared. It was found that OLP changed the expression profile of miR-155 and miR-19a, which in turn directly affected the production of eNOS and TLR2, respectively. In addition, by synergistically inducing an imbalance between Th1 and Th2, the simultaneous deregulation of miR-155/eNOS and miR-19a/TLR2 was responsible for an elevated risk of OLP.

A mechanistic linkage between oral lichen planus and autoimmune thyroid disease.

OBJECTIVE: To determine the levels of antithyroid antibodies and thyroid hormones in the sera of patients with oral lichen planus (OLP), and to quantify the expression of thyroid proteins in OLP lesions. SUBJECTS AND METHODS: Venous blood samples were drawn from 110 patients with OLP who had no history of thyroid disease or levothyroxine supplementation (OLP+/LT4 -). A random population sample of 657 healthy subjects was used as the control group. Two additional groups were used as comparators. Immunohistochemical and qPCR analyses were performed on tissue specimens collected from the patients with OLP and thyroid disease and healthy subjects. RESULTS: No association was found between the presence of antithyroid antibodies and OLP. More patients in the OLP+/LT4 - group showed high levels of thyroid-stimulating hormone and low levels of free thyroxine than were seen in the control group. Thyroid-stimulating hormone receptor was more highly expressed in the OLP lesions of patients with thyroid disease than in the healthy oral mucosa. CONCLUSIONS: A significant number of patients with OLP who are not previously diagnosed with thyroid disease have thyroid parameters that are compatible with hypothyroidism. The expression of thyroid-stimulating hormone receptor in OLP lesions suggests that mechanisms related to autoimmune thyroid disease are involved in the aetiology of OLP.