Gene Expression Levels Of Cytokines Research Articles

Improvement of Gut Barrier Function by Potato Anthocyanins Is Dependent on Gut Microbiota

Abstract Objectives Ulcerative colitis (UC) is characterized by chronic colonic inflammation, impaired barrier function and gut bacterial dysbiosis. Anthocyanin-containing potatoes have been shown to maintain the intestinal barrier function in colitic mice. However, the role of gut microbiota in the anti-colitic effects of anthocyanin-containing potatoes is not clear. This study evaluated the gut barrier protective efficacy of purple- and red-fleshed potatoes using a DSS-induced murine model of colitis with the intact and antibiotic-depleted microbiota. Methods Four-week-old mice (C57BL6) were randomized into four groups, receiving a standard control diet, or a 20% purple-/red-fleshed potato (PP/RP) supplemented diet. A broad-spectrum antibiotic cocktail was used to deplete the gut bacteria. After eight weeks, mice were treated with 2% DSS in their drinking water for five days to induce colitis. The polyphenol composition of potatoes was analyzed by LC-MS/MS. Intestinal permeability was measured using FITC-dextran. Colonic myeloperoxidase (MPO) activities were determined using the o-dianisidine dihydrochloride method. The fecal short-chain fatty acids (SCFA) content of mice was measured by GC-FID. RT-PCR was used to analyze the relative gene expression levels of cytokines and bacterial abundance. Results The predominant anthocyanin detected in RP was pelargonidin (93.13%). Whereas petunidin (75.57%) and malvidin (21.05%) were the two major anthocyanins present in PP. Administration of antibiotics resulted in a reduction in fecal SCFA. Antibiotic-treated mice were more prone to the DSS-induced increase in gut permeability. Anthocyanin-containing potato diets alleviated DSS-induced colonic damage, elevation in histology score and gut permeability only in non-antibiotic mice. RP alone suppressed the elevated colonic MPO activity and the gene expression levels of MUC2 in mice with intact gut microbiota. Conclusions In summary, anthocyanin-containing potatoes alleviated colitis-associated colonic epithelial damage and increased gut permeability in intact microbiota mice but not in microbiota-ablated mice. This suggests that the gut microbiome is essential for the gut barrier protective activity of anthocyanin-containing potatoes. Funding Sources USDA-NIFA awards 2016–67,017-29,285 and 2019–67,017-29,258.

Regular aerobic exercise activates PDGF-BB/PDGFR-β signaling and modulates the inflammatory-anti-inflammatory balance in diet-induced obese mice

ObjectiveRegular aerobic exercise induces cardioprotection by counteracting the obesity-associated inflammatory response, dyslipidemia. PDGF-BB/PDGFR-β signaling is established as a crucial mechanism of endothelial cell-cardiomyocyte communication and cardioprotection, but its physiological roles in response to obesity and regular aerobic exercise are unknown. MethodsThirty C57BL/6 mice were divided into three groups: a normal diet group, a high-fat diet group, and a high-fat diet plus aerobic exercise group. Glucose metabolic parameters, inflammation-related indicators, and blood lipids indicators were detected. In addition, gene expression levels of the inflammatory factors, PDGF-BB, PDGFR-β, PI3K, Akt, eNOS, and P53 in cardiac tissue were quantified. Morphological analysis was also used to quantify the magnitude of inflammation. ResultsHigh-fat diet (HFD) feeding resulted in adiposity, dyslipidemia, and low levels of cardioprotective factors such as APN and eNOS (P < 0.05), which were improved significantly by 8 weeks of aerobic exercise (P < 0.05). HFD feeding increased the gene expression levels of proinflammatory cytokines and decreased the gene expression levels of anti-inflammatory cytokines in cardiac tissue (P < 0.05), which was reversed by regular aerobic exercise (P < 0.05). In addition, HFD feeding suppressed the levels of the cardioprotective factors PDGF-BB and eNOS through PDGF-BB/PDGFR-β/PI3K/Akt/eNOS signaling in cardiac tissue, while regular aerobic exercise activated PDGF-BB/PDGFR-β/PI3K/Akt/eNOS signaling. ConclusionRegular aerobic exercise improved adiposity, dyslipidemia induced by HFD feeding. Regular aerobic exercise exerted a prominent role in modulating the inflammatory-anti-inflammatory balance and activating the levels of the cardioprotective factors eNOS and PDGF-BB through PDGF-BB/PDGFR-β signaling.

In vitro effects of curcumin on proinflammatory cytokines and expression of their genes in minor salivary gland tissue of patients with Sjogren's syndrome.

Curcumin reduces disease severity and ameliorates lupus-like/Sjögren's Syndrome-like disease in mice model. The immunological basis of these effects is largelyunknown. This study examined the effects of curcumin on pro-inflammatory cytokines secreted by minor salivary glands in patients with primary Sjögren's syndrome (pSS). Minor salivary gland (MSG) tissue samples were collected from patients undergoing biopsy for suspected pSS. The tissues were treated with phytohemagglutinin (PHA) alone as well as PHA with curcumin (30μM) and cultured in RPMI 1640 medium for 48h at 37°C in CO2 incubator. After the incubation period, culture supernatant and tissues were stored in the freezer (-80°C). IL-6 levels were measured in supernatant by ELISA kit. Gene expressions of pro-inflammatory cytokines, namely IL-6, IL-8, TNF-α, IL-1β, IL-4, IL-10, IL-17, IL-21, and IFN-γ, were measured by qPCR. IL-6 secretion levels and gene expressions were compared statistically between groups by Student's t test. Forty-seven patients were screened. Eight patients satisfied ACR/EULAR criteria for pSS. Seven patients with absent glandular inflammation and negative serology constituted sicca controls. These 15 subjects were included in final analysis. In pSS group, but not in controls, median IL-6 levels in supernatant were less in curcumin-treated as compared to PHA-alone subset [5.5 (0.7-13.34) vs 18.3 (12-32)ng/ml; p=0.0156]. mRNA expression levels of IL-6 were also lower in curcumin-treated samples as compared to PHA alone, when cases and controls were analyzed together as well as in cases alone (p=0.0009 and p=0.0078, respectively); however, mRNA expression of IL-1β was lower in curcumin-treated samples as compared to PHA alone, only when cases and controls were analyzed together (p=0.0215). There was no difference in other cytokine gene expression levels between the subsets under the in-vitro experimental conditions. In conclusion, curcumin reduced mRNA expression as well as secretion of IL-6 levels by salivary gland tissue of patients with pSS. Curcumin also suppressed PHA-induced mRNA expression levels of IL-6 and IL-1β in MSG tissue of patients with pSS and controls when analyzed together as a combined group.

Antiatherosclerotic Phenotype of Perivascular Adipose Tissue Surrounding the Saphenous Vein in Coronary Artery Bypass Grafting

BackgroundPerivascular adipose tissue (PVAT) is associated with metabolically driven chronic inflammation called metaflammation, which contributes to vascular function and the pathogenesis of vascular disease. The saphenous vein (SV) is commonly used as an essential conduit in coronary artery bypass grafting, but the long‐term patency of SV grafts is a crucial issue. The use of the novel “no‐touch” technique of SV harvesting together with its surrounding tissue has been reported to result in good long‑term graft patency of SV grafts. Herein, we investigated whether PVAT surrounding the SV (SV‐PVAT) has distinct phenotypes compared with other PVATs of vessels.Methods and ResultsFat pads were sampled from 48 patients (male/female, 32/16; age, 72±8 years) with coronary artery disease who underwent elective coronary artery bypass grafting. Adipocyte size in SV‐PVAT was significantly larger than the sizes in PVATs surrounding the internal thoracic artery, coronary artery, and aorta. SV‐PVAT and PVAT surrounding the internal thoracic artery had smaller extents of fibrosis, decreased gene expression levels of fibrosis‐related markers, and less metaflammation, as indicated by a significantly smaller extent of cluster of differentiation 11c–positive M1 macrophage infiltration, higher gene expression level of adiponectin, and lower gene expression levels of inflammatory cytokines, than did PVATs surrounding the coronary artery and aorta. Expression patterns of adipocyte developmental and pattern‐forming genes were totally different among the PVATs of the vessels.ConclusionsThe phenotype of SV‐PVAT, which may result from inherent differences in adipocytes, is closer to that of PVAT surrounding the internal thoracic artery than that of PVAT surrounding the coronary artery or that of PVAT surrounding the aorta. SV‐PVAT has less metaflammation and consecutive adipose tissue remodeling, which may contribute to high long‐term patency of grafting when the no‐touch technique of SV harvesting is used.

GM-1111 reduces radiation-induced oral mucositis in mice by targeting pattern recognition receptor-mediated inflammatory signaling.

PurposeOral mucositis (OM) is a common, painful side effect of radiation therapy used for the treatment of head and neck cancer (HNC). Activation of the innate immune system upon irradiation has been identified as a key precipitating event of OM. To better understand OM’s pathogenesis, we studied pattern recognition receptors (PRRs) and their downstream pro-inflammatory cytokines in a mouse model of radiation-induced OM. We also tested therapeutic efficacy of GM-1111 that targets innate immune system to reduce radiation-induced OM.Methods and materialsThe pathogenesis of OM was studied in a single X-ray induced mouse model. The severity of OM was measured by visual and microscopical examinations. The irradiation-induced changes of PRRs and their downstream effector cytokine gene expression levels were determined. The efficacy of GM-1111 to reduce OM was tested in single and fractionated irradiation mouse models. The impact of the drug on tumor response to radiation therapy was also tested in a mouse model of human HNC.ResultsRadiation-induced tissue ulcerations were radiation-dosage and -time dependent. The lesions showed selective increases in PRR and pro-inflammatory cytokine gene expression levels. Once daily administration of GM-1111 (≥30 mg/kg, s.c.) significantly reduced the severity and the incidence of OM. The drug had little effect on PRRs but significantly inhibited downstream pro-inflammatory cytokine genes. GM-1111 did not interfere radiation therapy to induce HNC SCC-25 tumor regression. Instead, we observed significant drug-induced tumor regression.ConclusionsRadiation induces tissue damages. The increased expression levels of PRRs and their downstream pro-inflammatory cytokine genes in the damaged tissues suggest their important contribution to the pathogenesis of OM. Drug GM-1111 that targets these innate immune molecules may be a potential drug candidate as an intervention for OM.

Secreted factors from dental pulp stem cells improve Sj\xf6gren\u2019s syndrome via regulatory T cell-mediated immunosuppression

BackgroundSjögren’s syndrome (SS) is a chronic autoimmune disease primarily characterized by inflammation in the salivary and lacrimal glands. Activated T cells contribute to disease pathogenesis by producing proinflammatory cytokines, which leads to a positive feedback loop establishment. The study aimed to evaluate the effects of secreted factors derived from dental pulp stem cells (DPSCs) or bone marrow mesenchymal stem cells (BMMSCs) on hyposalivation in SS and to investigate the mechanism involved.MethodsEighty percent confluent stem cells were replenished with serum-free Dulbecco’s modified Eagle’s medium and incubated for 48 h; following which, conditioned media from DPSCs (DPSC-CM) and BMMSCs (BMMSC-CM) were collected. Cytokine array analysis was performed to assess the types of cytokines present in the media. Flow cytometric analysis was performed to evaluate the number of activated T cells cultured in DPSC-CM or BMMSC-CM. Subsequently, DPSC-CM or BMMSC-CM was administered to an SS mouse model. The mice were categorized into the following groups (n = 6 each): non-treatment, Dulbecco’s modified Eagle’s medium (−), BMMSC-CM, and DPSC-CM. Histological analysis of the salivary glands was performed. The gene and protein expression levels of cytokines associated with T helper subsets in the submandibular glands (SMGs) were evaluated.ResultsDPSC-CM contained more secreted factors with tissue-regenerating mechanisms, such as cell proliferation, anti-inflammatory effects, and immunomodulatory effects. DPSC-CM was more effective in suppressing the activated T cells than other groups in the flow cytometric analysis. The stimulated salivary flow rate increased in SS mice with DPSC-CM compared with that in the other groups. In addition, the number of inflammation sites in SMGs of the mice administered with DPSC-CM was lower than that in the other groups. The expression levels of interleukin (Il)-10 and transforming growth factor-β1 were upregulated in the DPSC-CM group, whereas those of Il-4 and Il-17a were downregulated. The DPSC-CM-administered group presented with a significantly increased percentage of regulatory T (Treg) cells and a significantly decreased percentage of type 17 Th (Th17) cells compared with the other groups.ConclusionsThese results indicated that DPSC-CM ameliorated SS by promoting Treg cell differentiation and inhibiting Th17 cell differentiation in the mouse spleen.

Fermented milk with Lactobacillus curvatus SMFM2016-NK alleviates periodontal and gut inflammation, and alters oral and gut microbiota

This study aimed to analyze the effect of milk fermented with Lactobacillus curvatus SMFM2016-NK on periodontal diseases and gut health in a rat model. To improve the effect of Lb. curvatus SMFM2016-NK-fermented milk administration for relieving periodontitis, the periodontitis rat models were treated with the following for 4 wk: 10% skim milk (normal), periodontitis + 10% skim milk (negative control), periodontitis + Lactobacillus rhamnosus GG-fermented milk (positive control), and periodontitis + Lb. curvatus SMFM2016-NK-fermented milk (PD+LCFM). Transcriptional analysis of inflammatory cytokines [tumor necrosis factor α (TNF-α), IL-1β, IL-6, and IL-10] was performed via quantitative reverse-transcription PCR. The changes in the oral and gut microbiomes after administering Lb. curvatus SMFM2016-NK-fermented milk were analyzed with metagenomics sequencing using DNA extracted from the oral gingival tissues and feces from the cecum of the rat models. After treatment with Lb. curvatus SMFM2016-NK-fermented milk, the relative gene expression levels of TNFA and IL1B in the gingiva decreased in the PD+LCFM group compared with those in the negative control group. In the oral microbiome, the proportion of the phylum Proteobacteria in the PD+LCFM group was lower than that in the negative control after treatment with Lb. curvatus SMFM2016-NK-fermented milk. For the effect in the gut, the relative gene expression levels of inflammatory cytokines in the colon between the normal and negative control groups were not different; however, the expression levels of TNFA and IL1B in the PD+LCFM and positive control groups, respectively, were lower than those in the negative control group. The composition and diversity of the gut microbiome differed among normal, periodontitis, and Lb. curvatus SMFM2016-NK-fermented milk treatment groups. These results indicate that Lb. curvatus SMFM2016-NK-fermented milk could alleviate periodontal and gut inflammation and change oral and gut microbiota.

Systemic administration of \u03b2-glucan induces immune training in microglia

BackgroundAn innate immune memory response can manifest in two ways: immune training and immune tolerance, which refers to an enhanced or suppressed immune response to a second challenge, respectively. Exposing monocytes to moderate-to-high amounts of bacterial lipopolysaccharide (LPS) induces immune tolerance, whereas fungal β-glucan (BG) induces immune training. In microglia, it has been shown that different LPS inocula in vivo can induce either immune training or tolerance. Few studies focused on impact of BG on microglia and were only performed in vitro. The aim of the current study was to determine whether BG activates and induces immune memory in microglia upon peripheral administration in vivo.MethodsTwo experimental designs were used. In the acute design, mice received an intraperitoneal (i.p.) injection with PBS, 1 mg/kg LPS or 20 mg/kg BG and were terminated after 3 h, 1 or 2 days. In the preconditioning design, animals were first challenged i.p. with PBS, 1 mg/kg LPS or 20 mg/kg BG. After 2, 7 or 14 days, mice received a second injection with PBS or 1 mg/kg LPS and were sacrificed 3 h later. Microglia were isolated by fluorescence-activated cell sorting, and cytokine gene expression levels were determined. In addition, a self-developed program was used to analyze microglia morphological changes. Cytokine concentrations in serum were determined by a cytokine array.ResultsMicroglia exhibited a classical inflammatory response to LPS, showing significant upregulation of Tnf, Il6, Il1β, Ccl2, Ccl3 and Csf1 expression, three h after injection, and obvious morphological changes 1 and 2 days after injection. With an interval of 2 days between two challenges, both BG and LPS induced immune training in microglia. The training effect of LPS changed into immune tolerance after a 7-day interval between 2 LPS challenges. Preconditioning with BG and LPS resulted in increased morphological changes in microglia in response to a systemic LPS challenge compared to naïve microglia.ConclusionsOur results demonstrate that preconditioning with BG and LPS both induced immune training of microglia at two days after the first challenge. However, with an interval of 7 days between the first and second challenge, LPS-preconditioning resulted in immune tolerance in microglia.

Dental pulp-derived stem cell-conditioned media attenuates secondary Sjögren's syndrome via suppression of inflammatory cytokines in the submandibular glands.

IntroductionSjögren's syndrome (SS) is a chronic inflammatory autoimmune disease, which affects the exocrine glands. Its primary symptoms are decreased moisture in the mouth and eyes. Therapies are limited to treatment with steroids, which has unpleasant side effects, so new treatments would be beneficial. One possibility might be stem cells, such as bone marrow mesenchymal stem cells (BMMSCs) or dental pulp-derived stem cells (DPSCs); these have been reported to exert immunomodulatory effects on activated lymphoid cells. This study aimed to evaluate the effects of conditioned media from DPSCs (DPSC-CM) or BMMSCs (BMMSC-CM) on salivary functions in SS.MethodsCytokine array analysis was performed to assess the types of cytokines present in the media. DPSC-CM or BMMSC-CM was administered in an SS mouse model. Histological analysis of the salivary glands was performed, and gene expression levels of inflammatory and anti-inflammatory cytokines in the submandibular glands (SMGs) were evaluated.ResultsDPSC-CM contained more anti-inflammatory factors than BMMSC-CM. The mice that were given DPSC-CM had a lower number of inflammation sites in the SMGs than those in the other experimental groups, and their salivary flow rate increased. The expression levels of interleukin (IL)-10 and transforming growth factor-β1 increased in the DPSC-CM group, while those of Il-4, Il-6, and Il-17a decreased. The mice that received DPSC-CM showed a significantly increased percentage of regulatory T cells and a significantly decreased percentage of type T helper 17 cells compared to other groups.ConclusionsThese results indicate that DPSC-CM could be an effective therapy for SS-induced hyposalivation, since it decreases the number of inflammatory cytokines and regulates the local inflammatory microenvironment in the SMGs.

Familial Mediterranean fever-related miR-197-3p targets IL1R1 gene and modulates inflammation in monocytes and synovial fibroblasts

Familial Mediterranean fever (FMF); is an autosomal recessively inherited autoinflammatory disease caused by the mutations in the Mediterranean Fever (MEFV) gene. Recent studies have shown that epigenetic control mechanisms, particularly non-coding RNAs, may play a role in the pathogenesis of autoinflammation. microRNAs (miRNAs) are small non-coding RNAs that play critical roles in regulating host gene expression at the post-transcriptional level. The phenotypic heterogeneity of FMF disease suggests that FMF may not be a monogenic disease, suggesting that epigenetic factors may affect phenotypic presentation. Here we examined the potential anti-inflammatory effect of miR-197-3p, which is a differentially expressed miRNA in FMF patients, by using inflammation related functional assays. We monitored gene expression levels of important cytokines, as well as performed functional studies on IL-1β secretion, caspase-1 activation, apoptosis assay, and cell migration assay. These experiments were used to evaluate the different stages of inflammation following pre-miR-197 transfection. Anti-miR-197 transfections were performed to test the opposite effect. 3′UTR luciferase activity assay was used for target gene studies. Our results obtained by inflammation-related functional assays demonstrated an anti-inflammatory effect of miR-197-3p in different cell types (synovial fibroblasts, monocytes, macrophages). 3′UTR luciferase activity assay showed that miR-197-3p directly binds to the interleukin-1beta (IL-1β) receptor, type I (IL1R1) gene, which is one of the key molecules of the inflammatory pathways. This study may contribute to understand the role of miR-197-3p in autoinflammation process. Defining the critical miRNAs may guide the medical community in a more personalized medicine in autoinflammatory diseases.

Association between genetic polymorphism of IL-27 (rs153109) and toxoplasmosis in Iraqi women with recurrent abortion.

Single nucleotide polymorphisms (SNPs) are predictive markers for diseases, also cytokines are undergoing genetic controls and their genetic polymorphisms have a functional role in regulating the levels of cytokine gene expression. This study aims to reveal the association of toxoplasmosis with serum levels and SNP of IL-27 in aborted women. Total, 200 blood samples of patients and controls were collected from Al-Alawiya Maternity Teaching Hospital/Baghdad/Iraq from 2019–2020 for detecting the level of IL-27 by ELISA while the allelic discrimination method was used for SNP IL-27 (rs153109). The results indicated the IL-27 serum concentration elevated with significant differences in recurrent abortion with toxoplasmosis group compared to healthy women, pregnant women, and recurrent abortion. Also, recurrent abortion had significant differences compared to healthy women and pregnant women (P<0.05). Moreover, SNP results of IL-27 showed no significant association between patients and controls. Considering the distribution of serum levels for IL-27 by SNP, it was observed that IL-27 serum levels for TT, TC, and CC genotypes elevated in the patient group versus the control group. In addition, it was observed elevation serum level of IL-27 for the genotypes TT, TC, and CC in recurrent abortion with toxoplasmosis in contrast to healthy women, pregnant women, and recurrent abortion (P<0.05). Also, in recurrent abortion, the level of IL-27 for TC, and CC genotype showed significant differences comparing to healthy and pregnant women (P<0.05). In conclusion, the level of IL-27 in recurrent abortion women with toxoplasmosis was higher than the recurrent abortion women, which may be due to the inflammatory response to toxoplasmosis. SNP of IL-27 has not represented as a risk factor in recurrent abortion women with toxoplasmosis.

Analysis of the Relationship between the Expression Levels of Neutrophil Gelatinase-Associated Lipocalin and Cytokine Genes in Bone Marrow

Background: Recently, various associations of NGAL with several hematological cancers have been reported. However, given that the regulation of NGAL gene expression by cytokines is tissue-specific, NGAL expression in relation to those of cytokine genes has not been analyzed in bone marrow (BM) tissue. The purpose of this study was to analyze the association between NGAL and 48 cytokine gene expression levels in mononuclear cells (MNCs) of BM at the time of diagnosis of hematological malignancy and to explore the expression pattern of NGAL and related cytokine genes in patients with hematological malignancies and controls.Methods: BM MNCs were isolated from 48 patients, who were classified as patients presenting myeloproliferative neoplasm, acute myeloid leukemia, myelodysplastic syndrome, and as controls. NGAL and cytokine genes were analyzed using NanoString. Data on hematological parameters were collected from medical records. Single and multiple regression analyses were performed to analyze relationships.Results: Normalized counts of 26 cytokine genes were related to NGAL normalized counts, while STAT3 and TLR4 normalized counts had the highest explanatory power. The following multiple regression model was developed: NGAL normalized counts=4316.825 + 9.056 × STAT3 normalized counts + 844.226 × IL5 normalized counts + 17.540 × TLR1 normalized counts - 28.206 × TLR2 normalized counts - 42.524 × IRAK4 normalized counts. In the multiple regression analysis, STAT3 and TLR4 normalized counts showed multicollinearity. NGAL, STAT3, IL5, and TLR4 normalized counts showed similar intergroup patterns.Conclusions: NGAL normalized counts was predicted by a multiple regression model, while they showed similar intergroup patterns to STAT3, IL5, and TLR4 normalized counts.

Human umbilical cord mesenchymal stem cells ameliorate skin fibrosis development in a mouse model of bleomycin-induced systemic sclerosis.

Mesenchymal stem cell (MSC) infusion has become a novel therapeutic strategy for complex autoimmune diseases; however, few detailed studies have been performed to investigate the benefit and mechanism of MSC treatment on systemic sclerosis (SSc). The present study aimed to evaluate the therapeutic effect of human umbilical cord derived-MSCs (UC-MSCs) on bleomycin-induced SSc in mice and explore the potential underlying mechanism. The murine SSc model was established by daily subcutaneous injection of bleomycin for 4 weeks, followed with two UC-MSC infusions every 7 days. Skin fibrosis was assessed by H&E and Masson staining. Flow cytometry was used to determine IL-17A, IFN-γ, tumor necrosis factor-β, IL-10 and IL-12 levels in serum samples and T cell subsets in murine spleen. Additionally, gene expression levels of cytokines and fibrosis markers in skin samples were measured by reverse transcription-quantitative PCR. Immunofluorescence staining was performed to track UC-MSC localization and lymphocyte cell infiltration in vivo. UC-MSC treatment exerted an anti-fibrotic role in bleomycin-induced SSc mice, as confirmed by histological improvement, decreased collagen synthesis, and reduced collagen-1α1, collagen-1α2, fibronectin-1 and α-smooth muscle actin gene expression levels. The results indicated that UC-MSC treatment only had a limited systematic effect on cytokine production in serum samples and T cell activation in the spleen. By contrast, T helper (Th)17 cell infiltration and activation in skin were efficiently inhibited after UC-MSC infusion, as evidenced by the decreased IL-17A and retinoic acid-related orphan receptor γt gene expression as well as IL-17A production. UC-MSC administration significantly ameliorated bleomycin-induced skin fibrosis and collagen formation primarily by eliminating local inflammation and Th17 cell activation in the skin; however, the systemic inhibitory effect of UM-MSCs on cytokines was less profound.

CD71 mesangial IgA1 receptor and the progression of IgA nephropathy

The transferrin receptor (CD71) is known as a receptor for IgA1 on mesangial cells, but the role of CD71 in IgA nephropathy (IgAN) is unknown. We studied clinical implication of mesangial CD71 in 282 patients with biopsy-proven IgAN (2005-2018). The transcript and protein expression of glomerular CD71 was determined by real-time polymerase chain reaction and immunohistochemistry. Ten subjects with microscopic hematuria only and no evidence of histologic abnormalities on kidney biopsy were considered as controls. Human mesangial cells (HMCs) were treated with sera from IgAN patients and expression levels of CD71 and inflammatory cytokine markers were compared according to disease status. Disease progression was defined as a ≥30% decline in estimated glomerular filtration rate from the baseline value. During a mean follow up of 53.5 (18.3-75.9) months, 80 (28.4%) patients developed disease progression. The mRNA expression of CD71 was significantly higher in progressors than in nonprogressors (P = 0.001). Among the Oxford classification scores, patients with M1 had significantly higher CD71 expression levels than those with M0. In a multivariable Cox model, elevated transcript levels of CD71 were significantly associated with 4.32-fold higher risk of disease progression (P = 0.009). Furthermore, CD71 expression levels independently predicted the increase in proteinuria of ≥50% from the baseline (P = 0.03). Finally, HMCs treated with sera from IgAN patients with the higher Oxford score (M1E1S1T0) more increased the mRNA expression of CD71 and inflammatory markers than those with sera from negative score (M0E0S0T0). However, silencing CD71 significantly reduced expression levels of the inflammatory cytokine genes. Our results show that mesangial CD71 is significantly associated with disease progression and may play a biologic role in IgAN.

Streptococcus mutans bystander-induced bioeffects following sonodynamic antimicrobial chemotherapy through sonocatalytic performance of Curcumin-Poly (Lactic-co-Glycolic Acid) on off-target cells.

To assessed the Streptococcus mutans bystander-induced bioeffects following sonodynamic antimicrobial chemotherapy (SACT) by Curcumin-Poly (Lactic-co-Glycolic Acid) nanoparticles (Cur-PLGA-NPs). Cur-PLGA-NPs were synthesized and characterized by Scanning Electron Microscope (SEM), Transmission Electron Microscope (TEM), and Attenuated Total Reflection Fourier Transform IR (ATR-FTIR) spectroscopy, as well as, determination of in vitro drug release. Following the successful synthesis and characterization of Cur-PLGA-NPs, the cell survival, intracellular ROS production, apoptotic effects, DNA fragmentation, and gene expression levels of pro-inflammatory cytokines were investigated on human gingival fibroblast (HGF) cells as off-target cells through S. mutans bystander-induced bioeffects following SACT (BCSS). No significant cytotoxic and damage caused by the release of ROS from BCSS were observed in HGF cells (P > 0.05). There was no DNA fragmentation and anti-proliferation effects on HGF cells. The expression levels of bFGF, TNF-α, and IL-8 genes were increased after exposure to BCSS to 15.4-, 13.5-, and 8.7-fold, respectively (P < 0.05), while TGF-ß and IL-10 were downregulated to -4.1- and -6.8-fold, respectively (P < 0.05). It could be concluded that there were no bystander bioeffects of targeted sonocatalytic stress on off-target cells.

Lipid Pathology of the Corpus Callosum in Schizophrenia and the Potential Role of Abnormal Gene Regulatory Networks with Reduced Microglial Marker Expression.

Structural changes in the corpus callosum have been reported in schizophrenia; however, the underlying molecular mechanism remains unclear. As the corpus callosum is high in lipid content, we analyzed the lipid contents of the corpora callosa from 15 patients with schizophrenia and 15 age- and sex-matched controls using liquid chromatography coupled to tandem mass spectrometry and identified lipid combinations associated with schizophrenia. Real-time quantitative polymerase chain reaction analyses using extended samples (schizophrenia, n = 95; control, n = 91) showed low expression levels of lipid metabolism-related genes and their potential upstream transcription factors in schizophrenia. Subsequent pathway analysis identified a gene regulatory network where nuclear factor of activated T cells 2 (NFATC2) is placed most upstream. We also observed low gene expression levels of microglial markers, inflammatory cytokines, and colony-stimulating factor 1 receptor (CSF1R), which is known to regulate the density of microglia, in the corpus callosum in schizophrenia. The interactions between CSF1R and several genes in the presently identified gene network originating from NFATC2 have been reported. Collectively, this study provides evidence regarding lipid abnormalities in the corpora callosa of patients with schizophrenia and proposes the potential role of impaired “NFATC2-relevant gene network-microglial axis” as its underlying mechanism.

FGF21 alleviates neuroinflammation following ischemic stroke by modulating the temporal and spatial dynamics of microglia/macrophages

BackgroundResident microglia and macrophages are the predominant contributors to neuroinflammation and immune reactions, which play a critical role in the pathogenesis of ischemic brain injury. Controlling inflammatory responses is considered a promising therapeutic approach for stroke. Recombinant human fibroblast growth factor 21 (rhFGF21) presents anti-inflammatory properties by modulating microglia and macrophages; however, our knowledge of the inflammatory modulation of rhFGF21 in focal cerebral ischemia is lacking. Therefore, we investigated whether rhFGF21 improves ischemic outcomes in experimental stroke by targeting microglia and macrophages.MethodsC57BL/6 mice were subjected to middle cerebral artery occlusion (MCAO) and randomly divided into groups that received intraperitoneal rhFGF21 or vehicle daily starting at 6 h after reperfusion. Behavior assessments were monitored for 14 days after MCAO, and the gene expression levels of inflammatory cytokines were analyzed via qRT-PCR. The phenotypic variation of microglia/macrophages and the presence of infiltrated immune cells were examined by flow cytometry and immunostaining. Additionally, magnetic cell sorting (MACS) in combination with fluorescence-activated cell sorting (FACS) was used to purify microglia and macrophages.ResultsrhFGF21 administration ameliorated neurological deficits in behavioral tests by regulating the secretion of pro-inflammatory and anti-inflammatory cytokines. rhFGF21 also attenuated the polarization of microglia/macrophages toward the M1 phenotype and the accumulation of peripheral immune cells after stroke, accompanied by a temporal evolution of the phenotype of microglia/macrophages and infiltration of peripheral immune cells. Furthermore, rhFGF21 treatment inhibited M1 polarization of microglia and pro-inflammatory cytokine expression through its actions on FGF receptor 1 (FGFR1) by suppressing nuclear factor-kappa B (NF-κB) and upregulating peroxisome proliferator-activated receptor-γ (PPAR-γ).ConclusionsrhFGF21 treatment promoted functional recovery in experimental stroke by modulating microglia/macrophage-mediated neuroinflammation via the NF-κB and PPAR-γ signaling pathways, making it a potential anti-inflammatory agent for stroke treatment.

Co-exposure to different sized polystyrene microplastics and benzo[a]pyrene affected inflammation in zebrafish andbronchial-associated cells

In recent years, emerging research has revealed that microplastics (MPs) can pollute the air. Cellulose and plastic fibers have been found in the lungs of patients with lung cancer, indicating that MPs can “travel” through the atmosphere into the respiratory tract. Due to the hydrophobicity and relatively large surface area of MPs, the biotoxicity of MPs is greatly enhanced by their adsorption and enrichment in persistent organic pollutants (POPs). With the rapid development of the economy, emissions from coal burning and automobile exhaust have made polycyclic aromatic hydrocarbon (PAHs) one of the key pollutants in urban air that cannot be ignored. Epidemiological studies have shown that PAH exposure is associated with the development of respiratory diseases, such as bronchitis and asthma, in children. The aim of this study was to investigate the inflammatory effects of co-exposure to different size polystyrene MPs (PS-MPs) and benzo[a]pyrene (BaP) on adult zebrafish gills and bronchial-related cells to provide basic information for assessment of the potential health risks of combined exposure to the human respiratory system. PS-MPs (with size of 0.5 and 5 μm) and BaP were co-administered to embryos for 7 d, adult zebrafish for 14 d, and HBE cells and HBMSCs for 48 h. The relative expression levels of inflammatory cytokine genes in the different exposure groups were determined by real-time polymerase chain reaction (PCR). The ingestion of microplastics into fish bodies and cells was observed by stereoscope and confocal microscopy, respectively. Result shows that microplastics of two sizes could be ingested by zebrafish embryos. When exposed only to 0.5-μm PS-MPs, relative expression of the TNF-α , IL-6 , and IL-8 genes was significantly increased compared with that in the control; relative expression of the IFN-γ gene was largely increased in the gills of zebrafish in 5-μm PS-MP-exposed group. Moreover, relative expression of the IFN-γ , IL-6 and IL-8 genes in the group co-exposed to BaP and 0.5-μm PS-MPs was higher than that in BaP group. In addition, 0.5-μm PS-MPs could be ingested by both cell lines. Upon exposure to 0.5-μm PS-MPs alone, relative expression of genes TNF-α , IL-6 , and IL-8 was considerably increased compared with that in the control HBE cells. In HBSMCs, upon exposure to 0.5-μm PS-MPs alone, relative expression of the TNF-α , IL-8 , and IL-1β genes was substantially increased compared with that in the control group. Compared with that following exposure to BaP alone, relative expression of the IFN-γ and IL-6 genes in HBSMCs in 0.5-μm PS-MPs and BaP co-exposure group was remarkably increased, indicating a possible combined effect; however, co-exposure of 5-μm PS-MPs and BaP weakened the proinflammatory function of BaP and especially influenced relative expression of the TNF-α and IL-8 genes. In addition, none of the co-exposure groups of HBE cells showed obvious combined effects. The corollary is that particle size may affect the ingestion of PS-MPs in cells. In addition, the size-dependent effects of PS-MPs influence the accumulation of organic pollutants and their biotoxicity.

Renoprotective effects of paramylon, a β-1,3-D-Glucan isolated from Euglena gracilis Z in a rodent model of chronic kidney disease

Paramylon is a novel β-glucan that is stored by Euglena gracilis Z, which is a unicellular photosynthesizing green alga with characteristics of both animals and plants. Recent studies have indicated that paramylon functions as an immunomodulator or a dietary fiber. Currently, chronic kidney disease (CKD) is a global health problem, and there is no effective preventive treatment for CKD progression. However, paramylon may suppress the progression of CKD via the elimination of uremic toxins or modulation of gut microbiota, leading to the alleviation of inflammation. The aim of this study was to evaluate the effect of paramylon in CKD rat model. Eight-week-old male Wistar rats with a 5/6 nephrectomy were given either a normal diet or a diet containing 5% paramylon for 8 weeks. Proteinuria was measured intermittently. Serum and kidney tissues were harvested after sacrifice. We performed a renal molecular and histopathological investigation, serum metabolome analysis, and gut microbiome analysis. The results showed that paramylon attenuated renal function, glomerulosclerosis, tubulointerstitial injury, and podocyte injury in the CKD rat model. Renal fibrosis, tubulointerstitial inflammatory cell infiltration, and proinflammatory cytokine gene expression levels tended to be suppressed with paramylon treatment. Further, paramylon inhibited the accumulation of uremic toxins, including tricarboxylic acid (TCA) cycle-related metabolites and modulated a part of CKD-related gut microbiota in the CKD rat model. In conclusion, we suggest that paramylon mainly inhibited the absorption of non-microbiota-derived uremic solutes, leading to protect renal injury via anti-inflammatory and anti-fibrotic effects. Paramylon may be a novel compound that can act against CKD progression.

Evaluation of the Effectiveness of One- and Multi-Session Exposure-Based Treatments in Reducing Biological and Psychological Responses to Rat Phobia Among Students.

BackgroundAnimal phobia is one of the most common forms of specific phobias. This anxiety disorder challenges the medical student working with animal models. Regarding this, the present study was conducted to investigate the effectiveness of one- and multi-session cognitive exposure-based treatments in students with rat phobia.MethodsFor the purpose of the study, a total of 40 female students with rat phobia were allocated into two groups of one- and multi-session cognitive exposure-based treatments. The data were collected using psychological measures, including state anxiety, rat phobia, and disgust questionnaires, which were completed in three stages, including the baseline, pre-treatment, and post-treatment. The gene expression levels of pro-inflammatory cytokines (ie, interleukin-1 [IL-1], nuclear factor-kappaB [NF-κB], and tumor necrosis factor-alpha [TNFα]) associated with acute stress, as well as the serum levels of IL-6 and cortisol, were determined using reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) methods. This study was registered at the Iranian Registry of Clinical Trials (IRCT20171123037602N1).ResultsAccording to the results, both treatments yielded a significant reduction in almost all psychological measures and biological variables, except for IL-6. Rat phobia was the only variable that showed a statistically greater reduction in the multi-session treatment group. Furthermore, rat phobia and disgust reduction were maintained in both groups to the same extent during follow-up.ConclusionThe findings of the present study were indicative of the incidence of habituation in psychological and biological factors following exposure therapy. Both one- and multi-session treatments reduced the factors associated with rat phobia almost to the same degree. As a result of the high levels of disgust, anxiety-related biological factors remained high in four students despite observing a significant reduction in their fear. This led to passive avoidance in this group. The OST enabled the students to handle rats in less than half a day. Accordingly, it could be applied as a half-day workshop for students in medical universities to avoid the incidence of associated anxiety-related disorders in this group.