MicroRNA-155 expression is associated with pulpitis progression by targeting SHIP1.

BACKGROUND: Pulpitis is a commonly seen oral inflammation condition in clinical practice, it can cause much pain for the patient and may induce infections in other systems. Much is still unknown for the pathogenic mechanism of pulpitis. In this work, we discovered that the expression of miR-155 was associated with dental pulpal inflammation both in vivo and in vitro. METHODS AND RESULTS: Our experiments of LPS stimulated odontoblast cell line MDPC-23 showed miR-155 could act as a positive regulator by increasing the production of pro-inflammatory cytokines IL-1ß and IL-6 during inflammatory responses, whereas knockdown of miR-155 can reverse the effects. Bioinformatics analysis demonstrated that SHIP1 is a direct target of miR-155 in odontoblasts, this result was further verified at both mRNA and protein level. Inhibition of miR-155 resulted in the downregulation of inflammation factors, while co-transfection of si-SHIP1 and miR-155 inhibitor promoted the inflammatory responses. Treatment with miR-155 mimic or si-SHIP1 up-regulated the protein level of p-PI3K and p-AKT. By contrast, miR-155 inhibitor exerted the opposite effects. miR-155 mimics could upregulate the gene expression of IL-1ß and IL-6. Co-transfection of LY294002 and miR-155 mimic attenuated the inflammatory responses. Consistent with in vitro results, miR-155-/- mice could alleviate inflammatory response, as well as decrease the activation of p-PI3K and p-AKT, whereas increase the activation of SHIP1. CONCLUSIONS: Our data revealed a novel role for miR-155 in regulation of dental pulpal inflammatory response by targeting SHIP1 through PI3K/AKT signaling pathway.

Lactobacillus acidipiscis Induced Regulatory Gamma Delta T Cells and Attenuated Experimental Autoimmune Encephalomyelitis.

Multiple sclerosis is a chronic autoimmune disease involving the central nervous system, and shows a high disability rate. Its pathogenesis is complicated, and there is no good treatment. In recent years, with in-depth studies on the regulation of gastrointestinal flora, the relationship between the mammalian immune system and the intestinal flora has been extensively explored. Changes in the composition and structure of the gastrointestinal flora can affect the characteristics and development of the host immune system and even induce a series of central nervous system inflammation events. The occurrence and development of multiple sclerosis are closely related to the continuous destruction of the intestinal barrier caused by intestinal dysbacteriosis. In this study, we analyzed Lactobacillus acidipiscis in a mouse model of experimental autoimmune encephalomyelitis (EAE). We found that the amount of L. acidipiscis in the intestinal tract was inversely proportional to the progress of EAE development. In addition, the number of CD4+ FOXP3+ regulatory T cells in the mesenteric lymph nodes of mice increased significantly after the mice were fed with L. acidipiscis, and the differentiation of CD4+ T cells to Th1 and Th17 cells was inhibited. However, the protective effect of L. acidipiscis was lost in γδ T cell-deficient mice and hence was concluded to depend on the presence of regulatory γδ T cells in the intestinal epithelium. Moreover, including L. acidipiscis enhanced the development of Vγ1+γδ T cells but suppressed that of Vγ4+γδ T cells. In summary, our results demonstrated the ability of L. acidipiscis to induce generation of regulatory γδ T cells that suppress the development of the encephalomyelitic Th1 and Th17 cells and the progress of EAE.

Association of Polymorphisms in Inflammatory Cytokines Encoding Genes With Anti-N-methyl-D-Aspartate Receptor Encephalitis in the Southern Han Chinese.

Background: Single nucleotide polymorphisms (SNPs) that occur within genes encoding inflammatory cytokines can result in quantitative or qualitative changes in their expression or functionality, potentially leading to the development of anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. This study sought to evaluate the relationship between SNPs in inflammatory cytokines genes and the incidence of anti-NMDAR encephalitis in the Southern Han Chinese. Methods: In total, we enrolled 107 patients with anti-NMDAR encephalitis as well as 202 inpatient controls who had no first-degree relative with autoimmune diseases. Genotyping determination of all 309 patients was conducted for the IL-1ß rs16944, IL-4 rs2243250, IL-4 rs2070874, IL-6 rs1800796, IL-10 rs1800872, and IL-17 rs2275913 gene SNPs. Results: We observed statistically significant differences in the frequencies of G allele in IL-1ß rs16944 between anti-NMDAR encephalitis and controls (p = 0.017). Also, IL-1ß, IL-4, IL-6, IL-10, and IL-17 SNPs were not associated with the disease (p > 0.05). Conclusions: We found that patients with anti-NMDAR encephalitis exhibit a distinct immunological profile, and we found that the decreased frequency of G allele in IL-1ß rs16944 showed a protective role for anti-NMDAR encephalitis in the Southern Han Chinese.

Effect of ciliary neurotrophic factor on neural differentiation of stem cells of human exfoliated deciduous teeth.

The stem cells of human exfoliated deciduous teeth (SHEDs) are considered to be one of the main sources of seed cells in stem cell therapy. The aim of this study was to examine the effect of ciliary neurotrophic factor (CNTF) on neurogenic differentiation of SHEDs. With the consent of parents, SHEDs from 6 to 8 year old children were isolated and cultured. The mesenchymal stemness and the potential of multidirectional (adipogenic and osteogenic) differentiation for the isolated SHEDs were firstly determined. The effect of CNTF on specific neurogenic differentiation of SHEDs was then examined by detecting the expression of marker genes and proteins via RT-PCR, immunoblotting, and immunofluorescence microscopy. The isolated SHEDs expressed specific surface markers of mesenchymal stem cells, and their potential of osteogenic and adipogenic differentiation were confirmed. CNTF promoted the differentiation of SHEDs into neuron-like cells with a high expression of acetylcholine transferase (CHAT), a marker of cholinergic neurons. The expression of other neuron markers including nestin, microtubule-associated protein 2 (MAP 2), and ß-tublin III was also detected. Interestingly, the expression of neurogenic markers was maintained at a high level after neurogenic induction. SHEDs can be induced by CNTF to differentiate into cholinergic neuron-like cells under appropriate culture conditions. Our findings have laid a foundation for future use of SHEDs to treat neurological diseases.

Metformin enhances osteogenic differentiation of stem cells from human exfoliated deciduous teeth through AMPK pathway.

Stem cells from human exfoliated deciduous teeth (SHEDs) are ideal seed cells in bone tissue engineering. As a first-line antidiabetic drug, metformin has recently been found to promote bone formation. The purpose of this study was to investigate the effect of metformin on the osteogenic differentiation of SHEDs and its underlying mechanism. SHEDs were isolated from the dental pulp of deciduous teeth from healthy children aged 6 to 12, and their surface antigen markers of stem cells were detected by flow cytometry. The effect of metformin (10-200 µM) treatment on SHEDs cell viability, proliferation, and osteogenic differentiation was analyzed. The activation of adenosine 5'-monophosphate-activated protein kinase (AMPK) phosphorylation Thr172 (p-AMPK) was determined by western blot assay. SHEDs were confirmed as mesenchymal stem cells (MSCs) on the basis of the expression of characteristic surface antigens. Metformin (10-200 µM) did not affect the viability and proliferation of SHEDs but significantly increased the expression of osteogenic genes, alkaline phosphatase activity, matrix mineralization, and p-AMPK level expression in SHEDs. Compound C, a specific inhibitor of the AMPK pathway, abolished metformin-induced osteogenic differentiation of SHEDs. Moreover, metformin treatment enhanced the expression of proangiogenic/osteogenic growth factors BMP2 and VEGF but reduced the osteoclastogenic factor RANKL/OPG expression in SHEDs. In conclusion, metformin could induce the osteogenic differentiation of SHEDs by activating the AMPK pathway and regulates the expression of proangiogenic/osteogenic growth factors and osteoclastogenic factors in SHEDs. Therefore, metformin-pretreated SHEDs could be a potential source of seed cells during stem cell-based bone tissue engineering.

Overlapping Clinical Syndromes in Patients with Glial Fibrillary Acidic Protein IgG.

OBJECTIVE: The aim of this paper is to report 2 cases with overlapping syndromes in autoimmune glial fibrillary acidic protein (GFAP) astrocytopathy. METHODS: Antibodies were detected by indirect immunofluorescence assay. Patient data were analyzed retrospectively. RESULTS: One patient presented with overlapping neuromyelitis optica spectrum disorder (NMOSD) and positive GFAP-IgG and aquaporin-4-IgG. His main symptoms included vision loss, hiccups, fever, headache, and ataxia. High leukocyte count and protein levels were found in cerebrospinal fluid. Brain magnetic resonance imaging (MRI) revealed abnormalities in the hippocampus, midbrain, pons, medulla, and meninges. Characteristic radial enhancing patterns were seen. The other patient was a male with relapsing polychondritis (RP) and positive GFAP-IgG. His main manifestations were meningoencephalitis and dementia. MRI showed extensive abnormalities in the white matter around the ventricles, temporal lobe, and thalamus, with enhancement. Both patients responded well to the treatment with steroids and immunosuppressants. CONCLUSIONS: Although overlapping syndromes are rare, we report positive GFAP-IgG in 2 cases with NMOSD or RP. Both patients had clinical features of GFAP astrocytopathy, but diagnosis of the condition was very challenging because of the overlapping presentation.

Fabrication and characterization of strontium-hydroxyapatite/silk fibroin biocomposite nanospheres for bone-tissue engineering applications.

Osteoinductive bone filling biomaterials are in high demand for effective bone defect reconstruction. In this study, we aimed to design both organic and inorganic substances containing strontium-doped hydroxyapatite/silk fibroin (SrHA/SF) biocomposite nanospheres as an osteoinductive bone defect-filling biomaterial. SrHA/SF nanospheres were prepared with different concentration of Sr using ultrasonic coprecipitation method. The nanospheres were characterized using XRD, FTIR, SEM, TEM, ICP-AES and TGA. Solid and dense SrHA/SF nanospheres with 500-700 nm size and rough surfaces were synthesized successfully. Higher crystallinity and HA/SF phase were observed with the increase in Sr-concentration. The doping of different concentration of Sr did not affect the size and surface characteristics of the nanospheres. ICP-AES data showed that Sr/Ca ratio in SrHA/SF is very close to the nominal value. Nanospheres with higher concentration of Sr did not negatively affect the biocompatibility, but enhanced viability of mesenchymal stem cells (MSCs). Moreover, SrHA/SF nanospheres showed higher osteogenic differentiation potential compared to HA/SF nanospheres as indicated by the results from ALP staining, ALP activity, and Runx2, Alp, Col-1 and Opn gene expression assay in MSCs culture. Our findings suggest this novel design of biocompatible and osteoinductive SrHA/SF biocomposite nanospheres as a potential bone defect-filling biomaterial for bone regenerative applications.

Gradient sequential treatment of temporomandibular joint disorders / 口腔疾病防治

@#Temporomandibular joint disorder (TMD) is a common clinical disease in stomatology that occurs fre⁃ quently in young people, mostly in women, with an incidence of approximately 30%; its clinical manifestations include mandibular dysfunction, regional pain, and clicking noises around the temporomandibular joint (TMJ). Some patients have tinnitus, headache and other symptoms. With regard to the treatment procedures of TMD, a gradient sequential treatment model is currently preferred, each of which has strict indications. Generally, conservative treatment or nonin⁃ vasive treatment is preferred and is suitable for patients with dysfunction or mild organic disease. The second⁃choice minimally invasive treatment is suitable for patients who have failed conservative treatment or patients with mild organic disease. Finally, open surgery, which is suitable for patients who are not responsive to the first two treatments and show severe organic lesions, can be considered. The formulation of an open surgery treatment diagnosis and treatment plan should be personalized, led by doctors, and completed with the cooperation of patients. This article describes the“gradi⁃ ent sequential treatment”of temporomandibular joint disorders.

miR-140-5p regulates T cell differentiation and attenuates experimental autoimmune encephalomyelitis by affecting CD4+T cell metabolism and DNA methylation.

We previously demonstrated that decreased expression of miR-140-5p was associated with the progression of multiple sclerosis (MS) and miR-140-5p targeted STAT1 and interfered with the expression of IFN-γ. However, the underlying mechanisms how miR-140-5p regulated the differentiation of encephalomyelitic CD4+T cell remained unclear. In this study, we analyzed the levels of miR-140-5p in a mouse model of experimental autoimmune encephalomyelitis (EAE). We also analyzed the outcomes in response to either over- or under-expression of miR-140-5p. We found that the expression of miR-140-5p was inversely related to the progression of EAE. With the remission of the disease, the expression of miR-140-5p was restored to levels comparable to the control. The expression of miR-140-5p was downregulated in the encephalomyelitic CD4+T cells whereas enhanced expression of miR-140-5p inhibited the development of T helper type 1 (Th1) cell and significantly attenuated EAE. MiR-140-5p also caused hypermethylation of STAT1 and demethylation of GATA3. Furthermore, we found that miR-140-5p enhanced mitochondrial glycolysis in CD4+T cells with simultaneous activation of ATP activity. By blockage of the respiratory electron transport chain with the inhibitors of complex I and III, the effect of miR-140-5p on Th1 differentiation was blocked, which suggested a role for mitochondrial respiratory pathway in miR-140-5p-mediated inhibition of Th1 differentiation. In summary, our results demonstrated that the expression of miR-140-5p was negatively correlated with the progression of EAE and that miR-140-5p regulated Th1 differentiation via DNA methylation and mitochondrial respiratory pathway.

Blockade of the NLRP3/Caspase-1 Axis Ameliorates Airway Neutrophilic Inflammation in a Toluene Diisocyanate-Induced Murine Asthma Model.

Multiple studies have addressed the vital role of Nod-like receptor protein 3(NLRP3)/caspase-1/IL-1ß signaling in asthma. Yet, the role of NLRP3/caspase-1 in toluene diisocyanate (TDI)-induced asthma is still obscure. The aim of this study is to investigate the role of the NLRP3/caspase-1 axis in TDI-induced asthma. Using an established murine model of TDI-induced asthma as described previously, we gave the asthmatic mice a highly selective NLRP3 inhibitor, MCC950, as well as the specific caspase-1 inhibitors VX-765 and Ac-YVAD-CHO for therapeutic purposes. Airway resistance was measured and bronchoalveolar lavage fluid was analyzed. Lungs were examined by histology, immunohistochemistry, Western blotting, and flow cytometry. TDI exposure elevated the expression of NLRP3 and caspase-1 that was coupled with increased airway hyperresponsiveness (AHR), neutrophil-dominated cell infiltration, pronounced goblet cell metaplasia, extensive collagen deposition, and increased TH2/TH17 responses. Both VX-765 and Ac-YVAD-CHO effectively inhibited the activation of caspase-1 in TDI-asthmatic mice that was accompanied by dramatic attenuation of AHR, airway inflammation, and airway remodeling, in addition to a decreased TH2 response and lower levels of IL-18 and IL-1ß. MCC950 blocked the activation of NLRP3 and downregulated protein expression of caspase-1, IL-1ß, and IL-18 in TDI-exposed mice. Furthermore, MCC950 remarkably alleviated AHR, airway inflammation, airway remodeling, and significantly suppressed TH2/TH17 responses. These findings suggested that blockade of the NLRP3/caspase-1 axis effectively prevents the progression of TDI-induced asthma and could be used as therapeutic targets for asthmatics.

Research on the inhibitory effect of Baicalin on the human tongue squamous cell carcinoma cell line SCC15 / 口腔疾病防治

Objective@#To explore the inhibitory effect and possible mechanism of Baicalin on the human tongue squamous cell carcinoma cell line SCC15 and to provide a new idea and experimental basis for the clinical prevention and treatment of tongue squamous cell carcinoma.@*Methods @#SCC15 cells cultured in DMEM alone were used as the control group, and SCC15 cells cultured in 20 mg/mL baicalin solution were used as the baicalin group. Scratch tests and Transwell migration tests were performed to detect changes in cell migration ability, and flow cytometry was used to detect changes in the cell cycle. Western blotting was used to detect differences in the phosphorylation levels of signal transduction and transcription activator 3 (STAT3).@*Results @# Compared with the control group, the scratch test and the Transwell migration test showed that the cell migration ability of cells in the baicalin group was significantly decreased (t=4.927, P=0.008); flow cytometry showed that the number of cells of the baicalin group increased in the G0/G1 phase (t=9.893, P=0.001), decreased in the S phase (t=8.528, P=0.001), and decreased in the G2/M phase (t=3.550, P=0.024); Western blotting results showed that the STAT3 protein of SCC15 cells in the baicalin group decreased (t=3.550, P=0.024), and the phosphorylation level significantly decreased (t=8.262, P=0.001).@*Conclusion @#Baicalin inhibits the human tongue squamous cell carcinoma cell line SCC15, and its mechanism may be related to a decrease in STAT3 pathway phosphorylation activity.

Noninvasive vaccination against infectious diseases.

The development of a successful vaccine, which should elicit a combination of humoral and cellular responses to control or prevent infections, is the first step in protecting against infectious diseases. A vaccine may protect against bacterial, fungal, parasitic, or viral infections in animal models, but to be effective in humans there are some issues that should be considered, such as the adjuvant, the route of vaccination, and the antigen-carrier system. While almost all licensed vaccines are injected such that inoculation is by far the most commonly used method, injection has several potential disadvantages, including pain, cross contamination, needlestick injury, under- or overdosing, and increased cost. It is also problematic for patients from rural areas of developing countries, who must travel to a hospital for vaccine administration. Noninvasive immunizations, including oral, intranasal, and transcutaneous administration of vaccines, can reduce or eliminate pain, reduce the cost of vaccinations, and increase their safety. Several preclinical and clinical studies as well as experience with licensed vaccines have demonstrated that noninvasive vaccine immunization activates cellular and humoral immunity, which protect against pathogen infections. Here we review the development of noninvasive immunization with vaccines based on live attenuated virus, recombinant adenovirus, inactivated virus, viral subunits, virus-like particles, DNA, RNA, and antigen expression in rice in preclinical and clinical studies. We predict that noninvasive vaccine administration will be more widely applied in the clinic in the near future.

CD44 deletion leading to attenuation of experimental autoimmune encephalomyelitis results from alterations in gut microbiome in mice.

Dysbiosis in gut microbiome has been shown to be associated with inflammatory and autoimmune diseases. Previous studies from our laboratory demonstrated the pivotal role played by CD44 in the regulation of EAE, a murine model of multiple sclerosis. In the current study, we determined whether these effects resulted from an alteration in gut microbiota and the short-chain fatty acid (SCFA) production in CD44 knockout (CD44KO) mice. Fecal transfer from naïve CD44KO but not C57BL/6 wild type (CD44WT) mice, into EAE-induced CD44WT mice, led to significant amelioration of EAE. High-throughput bacterial 16S rRNA gene sequencing, followed by clustering sequences into operational taxonomic units (OTUs) and biochemical analysis, revealed that EAE-induced CD44KO mice showed significant diversity, richness, and evenness when compared to EAE-induced CD44WT mice at the phylum level, with dominant Bacteroidetes (68.5%) and low Firmicutes (26.8%). Further, data showed a significant change in the abundance of SCFAs, propionic acid, and i-butyric acid in EAE-CD44KO compared to EAE-CD44WT mice. In conclusion, our results demonstrate that the attenuation of EAE seen following CD44 gene deletion in mice may result from alterations in the gut microbiota and SCFAs. Furthermore, our studies also demonstrate that the phenotype of gene knock-out animals may be shaped by gut microbiota.

Intramuscular injection of exogenous leptin induces adiposity, glucose intolerance and fatty liver by repressing the JAK2-STAT3/PI3K pathway in a rat model.

Obesity, diabetes and fatty liver disease are extremely common in leptin-resistant patients. Dysfunction of leptin or its receptor is associated with obesity. The present study aimed to assess the effects of intramuscular injection of exogenous leptin or its receptor on fat deposition and leptin-insulin feedback regulation. Forty-five 40-day old female Sprague Dawley (SD) rats were injected thrice with leptin or its receptor intramuscularly. Adiposity and fat deposition were assessed by assessing the Lee's index, body weight, food intake, and total cholesterol, high density lipoprotein, low density lipoprotein, and triglyceride levels, as well as histological properties (liver and adipose tissue). Serum glucose, leptin, and insulin amounts were evaluated, and glucose tolerance assessed to monitor glucose metabolism in SD rats; pancreas specimens were analyzed immunohistochemically. Hypothalamic phosphorylated Janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), and phosphatidylinositol-3-kinase (PI3K) signaling, and hepatic sterol regulatory element binding protein-1 (SREBP-1) were qualified by Western blotting. Leptin receptor immunogen reduced fat deposition, increased appetite, and lowered serum leptin levels, enhancing STAT3 signaling in hypothalamus and down-regulating hepatic SREBP-1. In contrast, SD rats administered leptin immunogen displayed significantly increased body weight and fat deposition, with up-regulated SREBP-1, indicating adiposity occurrence. SD rats administered leptin immunogen also showed glucose intolerance, ß- cell reduction in the pancreas, and deregulation of JAK2-STAT3/PI3K signaling, indicating that Lep rats were at risk of diabetes. In conclusion, intramuscular injection of exogenous leptin or its receptor, a novel rat model approach, can be used in obesity pathogenesis and therapeutic studies.

TNF-α inhibits the migration of oral squamous cancer cells mediated by miR-765-EMP3-p66Shc axis.

Whereas TNF-α can facilitate the metastasis of oral squamous cancer cells (OSCC), whether it inhibits the metastasis is not clear so far. In this study, we demonstrated that high dose TNF-α at 100ng/mL could in vitro significantly inhibit the migration of two OSCC cell lines, CAL-27 and SCC-25. To explore the related mechanisms, we focused on the involvement of the microRNAs and found that TNF-α increased the expression of miR-765. The upregulation of miR-765 was attributed to the inhibition of the migration. We showed that miR-765 directly targeted EMP3 and suppressed its expression. We also found that the expression of EMP3 was much higher in human oral squamous cancer in compare with the surrounding normal tissue. Interestingly, p66Shc, a downstream molecule in the EMP3-related signaling pathway, was increased by TNF-α. We found that the overexpression of p66Shc could suppress the migration through the enhanced E-cadherin and ZO-1 signals. Either silencing the expression of EMP3 or enhancing the expression of miR-765 could upregulate the expression of p66Shc. Together, our results demonstrated that TNF-α inhibited the metastasis of oral squamous cancer cell through the miR-765-EMP3-p66Shc axis, which may provide new insights for the therapy of oral squamous cancer.

Pulpal Tissue Inflammatory Reactions after Experimental Pulpal Exposure in Mice.

INTRODUCTION: The purpose of this study was to establish a stable experimental mice pulpal inflammatory model and to evaluate inflammatory reactions of pulpal tissue after pulpal exposure. METHODS: Pulpal inflammation was induced in 80 C57BL/6 mice by occlusal exposure of the pulp of the maxillary first molar. The mice were sacrificed randomly at 0, 1, 6, 12, 24, 48, and 72 hours after pulpal exposure. Mice without pulpal exposure served as controls. Maxillary teeth were obtained and prepared for histologic analyses and real-time polymerase chain reaction analyses. RESULTS: As the duration of pulpal exposure increases, the inflammatory reaction is exacerbated. Within 6 to 12 hours after pulpal exposure, pulp tissues experienced red blood cell extravasation to the destruction of the odontoblast layer. After 24 hours, necrosis was observed in the pulpal tissue; until 72 hours, necrosis spread to the whole coronal pulpal tissue, and a large number of inflammatory cells were found in the radicular pulpal tissue. The results of histomorphologic scores have the same trend; samples from the 72-hour group possessed the highest score followed by samples from other groups (P < .01). The expression levels of inflammatory cytokines increased over the 72 hours, and there was a high rate of inflammatory cytokine expression at 6 and 12 hours after pulpal exposure. CONCLUSIONS: Our study represents a stable mice model for studying pulpal inflammation in vivo. Mouse pupal inflammation progresses rapidly, with dramatic changes evident in just a few hours.

Inverse correlation of expression of microRNA-140-5p with progression of multiple sclerosis and differentiation of encephalitogenic T helper type 1 cells.

The role of microRNA in the regulation of encephalitogenic T-cell development is of interest in understanding the pathogenesis of multiple sclerosis (MS). Direct binding of microRNAs to their target mRNAs usually suppresses gene expression and facilitates mRNA degradation. In this study, we observed that the expression of several microRNAs was significantly altered in patients with MS. Interestingly, the expression of miR-140-5p, among other microRNAs, was significantly decreased in the peripheral blood mononuclear cells of patients with MS, and this microRNA may regulate encephalitogenic T helper type 1 (Th1) cell differentiation. The expression level of miR-140-5p was inversely correlated with disease severity with greater reduction in relapsing disease compared with remitting disease. Transfection of synthetic miR-140-5p in peripheral blood mononuclear cells suppressed encephalitogenic Th1 differentiation. Signal transducer and activator of transcription 1 (STAT1) was the functional target of miR-140-5p - transfection of the synthetic miR-140-5p suppressed activation of STAT1 and the expression of its downstream target, T-bet. Our results suggested that miR-140-5p is probably involved in the regulation of encephalitogenic T cells in the pathogenesis of MS.

Resveratrol attenuates lipopolysaccharide-induced acute kidney injury by suppressing inflammation driven by macrophages.

SCOPE: Acute kidney injury (AKI) is the most frequent and serious complication in sepsis, a potentially deadly inflammatory response induced by bacterial, viral, or fungal infection. LPS-induced AKI is associated with an abnormal inflammatory response, including renal endothelial dysfunction and renal inflammation. Resveratrol, a natural phytoalexin with low toxicity and anti-inflammatory properties, is known to protect endothelial cells and modulate the immune response in sepsis. METHODS AND RESULTS: This study investigates the potential protective effects of resveratrol on AKI induced by LPS exposure of mice. Resveratrol was administered as a pre- and posttreatment, or as a posttreatment alone following LPS injection and compared to control groups. Resveratrol significantly improved kidney function and lowered serum and kidney tissue inflammatory cytokine levels. Consistently, resveratrol prevented endotoxin-induced disruption of endothelial cell permeability and inhibited inflammation of kidney tissue. Resveratrol treatment attenuated the effects of LPS on macrophages, with significant inhibition of activation, cytokine release, and Toll-like receptor 4 activation. Resveratrol treatment also resulted in decreased expression of iNOS, Bcl-2, and Bcl-xL in macrophages, which was linked with induction of apoptosis in macrophages. CONCLUSION: Our studies suggest that resveratrol might represent a novel therapeutic agent to prevent and treat sepsis-induced AKI.

The emerging role of leptin antagonist as potential therapeutic option for inflammatory bowel disease.

Inflammatory bowel disease (IBD) is a chronic relapsing immune-mediated inflammatory disorder that affects millions of people around the world. Leptin is a satiety hormone produced primarily by adipose tissue and acts both centrally and peripherally. Leptin has been shown to play a major role in regulating metabolism, which increases during IBD progression. Leptin mediates several physiological functions including elevated blood pressure, tumorogenesis, cardiovascular pathologies and enhanced immune response in many autoimmune diseases. Recent development of a leptin mutant antagonist that blocks leptin activity raises great hope and opens up new possibilities for therapy in many autoimmune diseases including IBD. To this end, preliminary data from an ongoing study in our laboratory on pegylated leptin antagonist mutant L39A/D40A/F41A (PEG-MLA) treatment shows an inhibition of chronic colitis in IL-10-/- mice. PEG-MLA effectively attenuates the overall clinical scores, reverses colitis-associated pathogenesis including a decrease in body weight, and decreases systemic leptin level. PEG-MLA induces both central and peripheral leptin deficiency by mediating the cellular immune response. In summary, after blocking leptin activity, the correlative outcome between leptin-mediated cellular immune response, systemic leptin levels, and amount of adipose tissue together may provide new strategies for therapeutic intervention in autoimmune diseases, especially for intestinal inflammation.

Leptin antagonist ameliorates chronic colitis in IL-10⁻/⁻ mice.

BACKGROUND: Although the etiology of two major forms of inflammatory bowel disease (IBD), Crohn's disease (CD) and ulcerative colitis (UC) are unknown and evidence suggests that chronic intestinal inflammation is caused by an excessive immune response to mucosal antigens. Previous studies support the role for TGF-ß1 through 3 in the initiation and maintenance of tolerance via the induction of regulatory T cells (Tregs) to control intestinal inflammation. Leptin, a satiety hormone produced primarily by adipose tissue, has been shown to increase during colitis progression and is believed to contribute to disease genesis and/or progression. AIM: We investigated the ability of a pegylated leptin antagonist (PG-MLA) to ameliorate the development of chronic experimental colitis. RESULTS: Compared to vehicle control animals, PG-MLA treatment of mice resulted in an (1) attenuated clinical score; (2) reversed colitis-associated pathogenesis including a decrease in body weight; (3) reduced systemic and mucosal inflammatory cytokine expression; (4) increased insulin levels and (5) enhanced systemic and mucosal Tregs and CD39⁺ Tregs in mice with chronic colitis. The percentage of systemic and mucosal TGF-ß1, -ß2 and -ß3 expressing CD4⁺ T cells were augmented after PG-MLA treatment. The activation of STAT1 and STAT3 and the expression of Smad7 were also reduced after PG-MLA treatment in the colitic mice. These findings clearly suggest that PG-MLA treatment reduces intestinal Smad7 expression, restores TGF-ß1-3 signaling and reduces STAT1/STAT3 activation that may increase the number of Tregs to ameliorate chronic colitis. CONCLUSION: This study clearly links inflammation with the metabolic hormone leptin suggesting that nutritional status influences immune tolerance through the induction of functional Tregs. Inhibiting leptin activity through PG-MLA might provide a new and novel therapeutic strategy for the treatment of IBD.