Levels Of Related Cytokines Research Articles

Ellagic acid effects on disease severity, levels of cytokines and T-bet, RORγt, and GATA3 genes expression in multiple sclerosis patients: a multicentral-triple blind randomized clinical trial.

Multiple sclerosis (MS) is a chronic autoimmune disease. Ellagic acid is a natural polyphenol and affects the fate of neurons through its anti-inflammatory and antioxidant properties. The present study aimed to investigate ellagic acid effects on disease severity, the expression of involved genes in the pathogenesis of MS, and the levels of related cytokines. The present study was a triple-blind clinical trial. Eligible patients were randomly assigned to two groups: Ellagic acid (25 subjects) for 12 weeks, receiving 180 mg of Ellagic acid (Axenic, Australia) and the control group (25 subjects) receiving a placebo, before the main meals. Before and after the study, the data including general information, foods intake, physical activity, anthropometric data, expanded disability status scale (EDSS), general health questionnaire (GHQ) and pain rating index (PRI), fatigue severity scale (FSS) were assessed, as well as serum levels of interferon-gamma (IFNγ), interleukin-17 (IL-17), interleukin-4 (IL-4) and transforming growth factor-beta (TGF-β), nitric-oxide (NO) using enzyme-linked immunoassay (ELISA) method and expression of T-box transcription factor (Tbet), GATA Binding Protein 3 (GATA3), retinoic acid-related orphan receptor-γt (RORγt) and Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes were determined using Real-Time Quantitative Reverse Transcription PCR (RT-qPCR) method. Ellagic acid supplementation led to a reduction in IFNγ, IL-17, NO and increased IL-4 in the ellagic acid group, however in the placebo group no such changes were observed (-24.52 ± 3.79 vs. -0.05 ± 0.02, p < 0.01; -5.37 ± 0.92 vs. 2.03 ± 1.03, p < 0.01; -18.03 ± 1.02 vs. -0.06 ± 0.05, p < 0.01, 14.69 ± 0.47 vs. -0.09 ± 0.14, p < 0.01, respectively). Ellagic acid supplementation had no effect on TGF-β in any of the study groups (p > 0.05). Also, the Tbet and RORγt genes expression decreased, and the GATA3 gene expression in the group receiving ellagic acid compared to control group significantly increased (0.52 ± 0.29 vs. 1.51 ± 0.18, p < 0.01, 0.49 ± 0.18 vs. 1.38 ± 0.14, p < 0.01, 1.71 ± 0.39 vs. 0.27 ± 0.10, p < 0.01). Also, ellagic acid supplementation led to significant decrease in EDSS, FSS and GHQ scores (p < 0.05), and no significant changes observed in PRI score (p > 0.05). Ellagic acid supplementation can improve the health status of MS patients by reduction of the inflammatory cytokines and Tbet and RORγt gene expression, and increment of anti-inflammatory cytokines and GATA3 gene expression.Clinical trial registration: (https://en.irct.ir/trial/53020), IRCT20120415009472N22.

Baseline levels of miR-223-3p correlate with the effectiveness of electroconvulsive therapy in patients with major depression

There is a strong medical need to develop suitable biomarkers to improve the diagnosis and treatment of depression, particularly in predicting response to certain therapeutic approaches such as electroconvulsive therapy (ECT). MicroRNAs are small non-coding RNAs that have the ability to influence the transcriptome as well as proteostasis at the systems level. Here, we investigate the role of circulating microRNAs in depression and response prediction towards ECT. Of the 64 patients with treatment-resistant major depression (MDD) who received ECT treatment, 62.5% showed a response, defined as a reduction of ≥50% in the MADRS total score from baseline. We performed smallRNA sequencing in blood samples that were taken before the first ECT, after the first and the last ECT. The microRNAome was compared between responders and non-responders. Co-expression network analysis identified three significant microRNA modules with reverse correlation between ECT- responders and non-responders, that were amongst other biological processes linked to inflammation. A candidate microRNA, namely miR-223-3p was down-regulated in ECT responders when compared to non-responders at baseline. In line with data suggesting a role of miR-223-3p in inflammatory processes we observed higher expression levels of proinflammatory factors Il-6, Il-1b, Nlrp3 and Tnf-α in ECT responders at baseline when compared to non-responders. ROC analysis of confirmed the diagnostic power of miR-223-3p demarcating ECT-responders from non-responder subjects (AUC = 0.76, p = 0.0031). Our data suggest that miR-223-3p expression and related cytokine levels could serve as predictors of response to ECT in individuals with treatment-resistant depressive disorders.

Extraction technology, components analysis and anti-inflammatory activity in vitro of total flavonoids extract from Artemisia anomala S. Moore

Artemisia anomala S. Moore exerts many pharmacological activities, including the removing of the blood stasis, relieving of the fever and analgesia, reducing the swelling and dampness. In this study, the extraction technology, chemical compositions and anti-inflammatory effect in vitro and mechanism of total flavonoids extract from Artemisia anomala S. Moore were studied. The optimal yield rate of total flavonoids extract was optimized by single factor experiments and response surface method, and the chemical constituents were analyzed by UPLC-QTOF-MS method; and the anti-inflammatory activity of the extract was evaluated with lipopolysaccharide induced RAW 264.7 cells. The highest extraction rate was 2.02% under these conditions of the concentration of ethanol 50%, the ultrasonic extraction time 30 min, and the ratio of solvent volume to material weight 20:1 (ml/g). In addition, the main components of total flavonoid extract were preliminarily identified and deduced based on mass spectrometry information and relevant literatures, and its stronger anti-inflammatory activity was demonstrated by reducing the phagocytosis, the content of nitric oxide and the level of related cytokines (tumor necrosis factor-α, interleukin−10, interleukin-6). Furthermore, it was further revealed that the anti-inflammatory effect of the extract was closely connected with the activation of TLR4-MyD88-NF-κB signalling pathway. This study indicated that the total flavonoids extract from Artemisia anomala S. Moore may be a better candidate anti-inflammatory natural medicine.

P-370 Impaired endometrial decidualization with a hyperinflammation environment is involved in poor reproductive outcomes in adenomyosis patients

Abstract Study question Do patients with adenomyosis have an impaired inflammatory state of the endometrium that could affect implantation and pregnancy? Summary answer Adenomyosis patients show increased proinflammatory cytokines expression and deregulated decidualization markers expression in eutopic endometrium which could lead to altered endometrial receptivity and impaired implantation. What is known already Adenomyosis is an estrogen-dependent chronic inflammatory condition, characterized by the presence of endometrial glands and stroma within the myometrium. Adenomyosis patients present altered decidualization and defective embryo-endometrium communication, resulting in implantation failure, miscarriage, and other fertility-related disorders. Although the underlying mechanisms of this infertility remain unknown, it is known that a favorable immune and inflammatory uterine environment is necessary for developmental pregnancy. It has been proposed that a uterine hyperinflammatory state could be involved in adenomyosis-related infertility. For this reason, we aim to evaluate the inflammatory and endometrial receptivity status during implantation in eutopic endometrium from adenomyosis patients. Study design, size, duration Sixteen endometrial samples were collected from infertile patients with and without adenomyosis undergoing hormonal replacement therapy before in vitro fertilization (IVF) at IVIRMA Valencia between January to December 2022. Participants/materials, setting, methods Eutopic endometrial samples were obtained at secretory phase (LH + 7) from patients diagnosed with adenomyosis (n = 8) by ultrasound or hysteroscopy and patients without gynecological diseases (control, n = 8). Total RNA extraction was performed to later determine the gene expression of the decidualization-related genes Prolactin (PRL), SPP1 and PAEP by qRT-PCR. In addition, proteins were extracted from eutopic endometrium using a lysis buffer and the relative expression levels of human cytokines were measured by Human Cytokine Array (Raybiotech). Main results and the role of chance Gene expression evaluation in endometrium from adenomyosis patients compared to control showed a significant downregulation of the key marker of decidualization PRL (fold change [fc] = 0.56, p = 0.0047) and a significant upregulation of SPP1 (fc = 1.75, p = 0.009) and PAEP (fc = 1.56, p = 0.0192), both involved in endometrial receptivity and in immune regulation. Regarding cytokines expression, array results showed upregulation of different interleukins (IL) involved in the mediation of the immune and inflammatory response previously described in adenomyosis. Specifically, IL1ß (12±19.63 vs. 1.37±2.77, p = 0.009) that regulates several inflammatory responses, including cell proliferation, differentiation, and apoptosis; and IL6 (584.7±121.8 vs. 488.4±57.44, p = 0.06) that acts on the inflammation and maturation of B cells, contributing to the development of autoimmune diseases. Similarly, cytokines related to the promotion of inflammation and cellular proliferation in endometriosis, IL17a (173.6±44.75 vs. 102.10±71.85, p = 0.04) and TNFβ (430.9±139.6 vs. 315.3±37.09, p = 0.04), were also upregulated in adenomyosis compared to control. There were also other cytokines upregulated with diverse functions like an anti-inflammation response, growth factors or immune modulation: IL5 (308.1±88.77 vs. 202.6±45.72, p = 0.01), IL2 (341.4±60.38 vs. 270.8±35.13, p = 0.014), TGFα (215.3±27.04 vs. 117.8±36.67, p = 0.002), IL31 (77.74±40.57 vs. 14±15.96, p = 0.0012), IL7 (739.1±147.1 vs. 511.8±103.7, p = 0.003) and IL15 (301.9±95.73 vs. 209.8±50.62, p = 0.03). Limitations, reasons for caution Our findings are limited by the relatively small sample size and inherent biological variability of human samples. Wider implications of the findings Adenomyosis patients showed impaired decidualization and a hyperinflammatory endometrial environment that could be affecting the endometrial receptivity and consequently, the embryo implantation. These findings open insight to further investigations to study the mechanism by which exaggerated inflammation affects fertility in the context of adenomyosis to define new management approaches. Trial registration number Not applicable

Loricrin and Cytokeratin Disorganisation in Severe Forms of Periodontitis

ObjectiveThe aim of this research was to investigate the role of the cornified epithelium, the outermost layer of the oral mucosa, engineered to prevent water loss and microorganism invasion, in severe forms of periodontitis (stage III or IV, grade C). MethodsPorphyromonas gingivalis, a major periodontal disease pathogen, can affect cornified epithelial protein expression through chronic activation of signal transducer and activator of transcription 6 (Stat6). We used a mouse model, Stat6VT, that mimics this to determine the effects of barrier defect on P gingivalis–induced inflammation, bone loss, and cornified epithelial protein expression, and compared histologic and immunohistologic findings with tissues obtained from human controls and patients with stage III and IV, grade C disease. Alveolar bone loss in mice was assessed using micro-computerised tomography, and soft tissue morphology was qualitatively and semi-quantitatively assessed by histologic examination for several proteins, including loricrin, filaggrin, cytokeratin 1, cytokeratin 14, a proliferation marker, a pan-leukocyte marker, as well as morphologic signs of inflammation. Relative cytokine levels were measured in mouse plasma by cytokine array. ResultsIn the tissues from patients with periodontal disease, there were greater signs of inflammation (rete pegs, clear cells, inflammatory infiltrates) and a decrease and broadening of expression of loricrin and cytokeratin 1. Cytokeratin 14 expression was also broader and decreased in stage IV. P gingivalis–infected Stat6VT mice showed greater alveolar bone loss in 9 out of 16 examined sites, and similar patterns of disruption to human patients in expression of loricrin and cytokeratins 1 and 14. There were also increased numbers of leukocytes, decreased proliferation, and greater signs of inflammation compared with P gingivalis–infected control mice. ConclusionsOur study provides evidence that changes in epithelial organisation can exacerbate the effects of P gingivalis infection, with similarities to the most severe forms of human periodontitis.

Perilla, sunflower, and tea seed oils as potential dietary supplements with anti-obesity effects by modulating the gut microbiota composition in mice fed a high-fat diet.

Obesity has become a serious public health problem with its alarmingly increasing prevalence worldwide, prompting researchers to create and develop several anti-obesity drugs. Here, we aimed to investigate the protective effects of perilla seed oil (PSO), sunflower oil (SFO), and tea seed oil (TSO) against obesity through the modulation of the gut microbiota composition and related metabolic changes in mice fed a high-fat diet (HFD). Mice were divided into six equal groups: ND (normal diet); HFD; ORL (HFD supplemented with 20mg/kg body weight of orlistat); PSO, SFO, and TSO (HFD supplemented with 2g/kg body weight of PSO, SFO, and TSO, respectively). Our findings showed that PSO, SFO, and TSO supplementation significantly reduced body weight, organ weight, blood glucose, lipopolysaccharides (LPS), insulin resistance, and improved serum lipid levels (TG, TC, LDL-C, and HDL-C). Meanwhile, the three treatments alleviated oxidative stress and hepatic steatosis and reduced liver lipid accumulation. Relative mRNA expression levels of inflammatory cytokines (TNF-α, IL-1β, IL-6, and MCP-1) and lipid synthesis-related genes (PPAR-γ, FAS, and SREBP-1) were down-regulated, while β-oxidation-related genes (PPAR-α, CPT1a, and CPT1b) were up-regulated in the liver tissue of treated mice. Besides, dietary oil supplementation alleviated HFD-induced gut microbiota dysbiosis by promoting gut microbiota richness and diversity, decreasing the Firmicutes-to-Bacteroidetes ratio, and boosting the abundance of some healthy bacteria, like Akkermansia. PSO, SFO, and TSO supplementation could alleviate inflammation, oxidative stress, and hepatic steatosis, likely by modulating the gut microbiota composition in HFD-fed mice.

Geniposidic acid attenuates DSS-induced colitis through inhibiting inflammation and regulating gut microbiota.

Geniposidic acid (GPA) is a bioactive compound isolated from Gardenia jasminoides Ellis (Rubiaceae) that has long been used to treat arthritis, jaundice, and hypertension. However, the therapeutic effects of GPA against colitis remain underexplored. This study aimed to investigate the effect of GPA on the remission of colitis and the underlying mechanisms. A DSS-induced colitis mouse model was used to evaluate the influence of GPA on the modulation of gut microbiota and intestinal epithelial barrier function. Our results indicated that GPA improved DSS-induced mouse colitis, including loss of body weight, disease activity index (DAI), colon length, and colonic pathological damage. DSS-induced destruction of the intestinal barrier was also significantly repaired by GPA treatment. In addition, the relative levels of pro-inflammatory cytokines, such as IL-1β and TNF-α, were markedly alleviated by GPA. Furthermore, western blot analysis revealed that GPA downregulated the protein expression of the nuclear transcription factor NF-κB. Finally, we first demonstrated that GPA could alleviate gut microbiota dysbiosis in mice with colitis by bacterial 16S rRNA sequencing. In conclusion, our study demonstrates the therapeutic and protective effects of GPA on IBD and provides novel insights into the prevention of colitis by targeting gut microbiota metabolism using natural products.

Th1 promotes M1 polarization of intestinal macrophages to regulate colitis-related mucosal barrier damage.

This work aimed to investigate the role of helper T cell 1 (Th1) in chronic colitis and its immunoregulatory mechanism. The proportions of Th1 and Th2, and the levels of related cytokines in tissues from patients with inflammatory bowel disease (IBD; ulcerative colitis+Crohn's disease, UC+CD) were detected. DSS was used to induce the mouse model of IBD; thereafter, Th1 cells were induced in vitro and amplified before they were injected intraperitoneally. Later, the changes in life state and body weight of mice were observed, the proportion of M1 macrophages in mucosal tissues and mucosal barrier damage were detected. After treatment with macrophage scavenging agent (Clodronate Liposomes, CLL), the influence of Th1 on IBD mice was observed. Then, the intestinal macrophages were co-cultured with Th1 in vitro to observe the influence of Th1 on the polarization of intestinal macrophages. Besides, cells were treated with the STAT3 inhibitor to further detect the macrophage polarization level. Intestinal macrophages were later co-cultured with intestinal epithelial cells to observe the degree of epithelial cell injury. The Th1 proportions in intestinal tissues of UC and CD patients were higher than those in healthy subjects, but the difference in Th2 proportion was not significant. In the IBD mouse model, Th1 induced the M1 polarization of macrophages, aggravated the intestinal inflammatory response, and resulted in the increased mucosal barrier permeability. Pretreatment with CLL antagonized the effect of Th1 cells, reduced the intestinal tissue inflammatory response and mucosal barrier permeability.

KW2449 ameliorates collagen-induced arthritis by inhibiting RIPK1-dependent necroptosis.

Necroptosis has recently been found to be associated with the pathogenesis of many autoimmune diseases, including rheumatoid arthritis (RA). This study was undertaken to explore the role of RIPK1-dependent necroptosis in the pathogenesis of RA and the potential new treatment options. The plasma levels of receptor-interacting protein kinase 1 (RIPK1) and mixed lineage kinase domain-like pseudokinase (MLKL) in 23 controls and 42 RA patients were detected by ELISA. Collagen-induced arthritis (CIA) rats were treated with KW2449 by gavage for 28 days. Arthritis index score, H&E staining, and Micro-CT analysis were used to evaluate joint inflammation. The levels of RIPK1-dependent necroptosis related proteins and inflammatory cytokines were detected by qRT-PCR, ELISA and Western blot, and the cell death morphology was detected by flow cytometry analysis and high-content imaging analysis. The plasma levels of RIPK1 and MLKL in RA patients were higher than those in healthy people, and were positively correlated with the severity of RA. KW2449 could reduce joint swelling, joint bone destruction, tissue damage, and the plasma levels of inflammatory cytokines in CIA rats. Lipopolysaccharide combined with zVAD (LZ) could induce necroptosis in RAW 264.7 cells, which could be reduced by KW2449. RIPK1-dependent necroptosis related proteins and inflammatory factors increased after LZ induction and decreased after KW2449 treatment or knockdown of RIPK1. These findings suggest that the overexpression of RIPK1 is positively correlated with the severity of RA. KW2449, as a small molecule inhibitor targeting RIPK1, has the potential to be a therapeutic strategy for RA treatment by inhibiting RIPK1-dependent necroptosis.

Matrine alleviates depressive-like behaviors via modulating microbiota–gut–brain axis in CUMS-induced mice

BackgroundThe realization of the “microbiota–gut–brain” axis plays a critical role in neuropsychiatric disorders, particularly depression, is advancing rapidly. Matrine is a natural bioactive compound, which has been found to possess potential antidepressant effect. However, the underlying mechanisms of regulation of the “microbiota–gut–brain” axis in the treatment of depression by oral matrine remain elusive.MethodsIts antidepressant effects were initially evaluated by behavioral tests and relative levels of monoamine neurotransmitters, and matrine has been observed to attenuate the depression-like behavior and increase neurotransmitter content in CUMS-induced mice. Subsequently, studies from the “gut” to “brain” were conducted, including detection of the composition of gut microbiota by 16S rRNA sequencing; the metabolomics detection of gut metabolites and the analysis of differential metabolic pathways; the assessment of relative levels of diamine oxidase, lipopolysaccharide, pro-inflammatory cytokines, and brain-derived neurotrophic factor (BDNF) by ELISA kits or immunofluorescence.ResultsMatrine could regulate the disturbance of gut microbiota and metabolites, restore intestinal permeability, and reduce intestinal inflammation, thereby reducing the levels of pro-inflammatory cytokines in peripheral blood circulation and brain regions, and ultimately increase the levels of BDNF in brain.ConclusionMatrine may ameliorate CUMS-induced depression in mice by modulating the “microbiota–gut–brain” axis.

Levels of IL-6 in peripheral blood and cerebrospinal fluid of Alzheimer's disease: a meta-analysis

BACKGROUND: Alzheimer's disease (AD) is a common degenerative disease of the central nervous system in the elderly. Several reports have described the serum levels of interleukin (IL-6) in AD. In order to elucidate the status of IL-6 in AD, the article conducted a meta-analysis of previously published results to assess the levels of IL-6 from different tissue sources in AD. METHODS: The levels of Th17 related cytokines (IL-6) in peripheral blood or cerebrospinal fluid (CSF) of AD were systematically reported in domestic and foreign databases. The article assessed pooled data using a random-effects model.The I-square index (I2) was used to assess inconsistency and publication bias was assessed by examining funnel plot asymmetry using the Begg and Egger test. Results: Among 1348 identified studies, the article selected 45 studies for analysis, 36 to investigate IL-6 levels in peripheral blood and 9 to investigate IL-6 levels in CSF of AD versus healthy controls (HCs). The analysis revealed that IL-6 content in peripheral blood of AD was significantly higher than that of HCs[SMD = 0.913, 95% CI (0.345, 1.481), P < 0.05], while the level of IL-6 in CSF showed no statistical significance [SMD = 0.132, 95% CI (-0.579, 0.843), P > 0.05] and could not be used as a marker in AD. CONCLUSIONS: In AD, the level of IL-6, a relevant marker, is increased, and spontaneous IL-6 production by peripheral blood mononuclear cells may be a marker of future AD risk in the elderly, and these data strengthen evidence of the pathophysiological role of inflammation in the development of clinical AD, but suggest that the mechanism of IL-6 action still needs more research and can be used as a potential target to address the problem of AD in the future.

Iridoid from Eucommia ulmoides Oliv. Exerts Antiarthritis Effects by Inhibiting the JAK2/STAT3 Signaling Pathway In Vivo and In Vitro.

The purpose of this study was to investigate the anti-inflammatory effects of EU-Idd both in vivo and in vitro. In vivo, we used the collagen-induced arthritis (CIA) rat model to investigate the efficacy of EU-Idd on rheumatoid arthritis. Hematoxylin-eosin staining and Safranin O-fast green staining were used to evaluate the pathological status of the ankle joints in CIA rats. Micro-CT scanning was used to investigate bone erosion of the ankle joints. In vitro, the effect of EU-Idd on Th17 cell differentiation was identified by flow cytometry. TRAP staining was used to detect osteoclast cells. HFLS-RA model cells, induced by tumor necrosis factor-α(TNF-α), were used to evaluate the anti-inflammatory effects of EU-Idd while the levels of related inflammatory cytokines and JAK2/STAT3 proteins were detected by RT-qPCR and western blotting. EU-Idd alleviated joint inflammation in CIA rats and exerted protective effects on the ankle joints. EU-Idd also prevented the differentiation of CD4+ T cells into Th17 cells, reduced the number of osteoclasts, and improved the expression levels of bone metabolism-related proteins including OPG and RANKL. Moreover, EU-Idd inhibited the invasion and migration of HFLS-RA cells and downregulated the expression of related inflammatory cytokine genes and the protein expression levels of p-JAK2 and p-STAT3, both in vivo and in vitro. EU-Idd exerts anti-inflammatory and osteoprotective effects by regulating the JAK2/STAT3 pathway in rheumatoid arthritis. These results are beneficial to excavate new pharmaceutical ingredients for rheumatoid arthritis from iridoid.

PPARγ/RAD21 alleviates peripheral secondary brain injury in rat cerebral hemorrhage model through promoting M2 polarization of microglial cells

PPARγ has been reported to participate in intracerebral hemorrhage (ICH) progression, and recruit RAD21 through binding DNA. Our study aimed to explore the roles of PPARγ/RAD21 in ICH and their related mechanisms. ICH models in vitro and in vivo were established using thrombin and autologous blood injection, respectively. After that, rosiglitazone (RSG), GW9662, and RAD21 knockdown/overexpression plasmids were used to treat the ICH models. The cell apoptosis, the related inflammatory cytokines levels, and the neurological function of the rats were examined. Real-time quantitative PCR (RT-qPCR), western blot and immunofluorescence were employed to determine the expression of the M1/M2 polarization-related markers. Finally, the interaction of PPARγ and RAD21 in microglial cells was observed using double labeled immunofluorescence and co-immunoprecipitation. After thrombin induction, the cell apoptosis, and TNF-α, IL-1β and IL-10 contents were all significantly increased (P<0.05); whereas RSG and RAD21 overexpression evidently inhibited the apoptosis of thrombin-caused microglial cells, reduced TNF-α and IL-1β contents, further increased IL-10 content (P<0.05). The combination of RAD21 and PPARγ was enhanced by RSG and RAD21 overexpression. In vivo experiments showed that RSG and RAD21 overexpression decreased neurological deficit score, brain water content and hematoma volume. Additionally, RSG and RAD21 overexpression up-regulated the expression of PPARγ, RAD21, Arg1, KLF4, and TGF-β, whereas down-regulated iNOS and CD32 expression. The actions of GW9662 and RAD21 knockdown were opposite to those of RSG and RAD21 overexpression. PPARγ/RAD21 may alleviate ICH progression through promoting M2-type polarization of microglial cells and inhibiting inflammatory response.

Carbon monoxide-releasing molecule 2 inhibits inflammation associated with intestinal ischemia-reperfusion injury in a rat model of hemorrhagic shock.

Intestinal ischemia-reperfusion injury (IRI) occurs in multiple clinical settings and contributes to systemic inflammatory response syndrome (SIRS) and multiple organ dysfunction syndrome (MODS). Due to the innate inflammatory immune nature, T cells play a crucial role in the pathogenesis of IRI, which includes not only CD4+T cells, but also CD8+and γδ T cells. Carbon monoxide (CO) plays an important role in regulating CD4+T cell responses and has been proven to be a novel therapeutic target in a variety of inflammatory disease models. This study aimed to assess whether pretreatment with carbon monoxide-releasing molecule-2 (CORM-2) could ameliorate inflammation by regulating differentiation of CD4+T cells in intestinal mucosa of rats undergoing hemorrhagic shock. A hemorrhagic shock model was established to study intestinal IRI. Morphological changes were investigated using light microscopes. Fluorescein isothiocyanate-dextran (FITC-dextran) was used as an indicator of intestinal paracellular permeability. Transcription factors involved in differentiation of CD4+T cells in intestinal mucosa were detected by immunofluorescence, and the expression levels of related cytokines were determined by Western blotting. The results of hematoxylin-eosin (H-E) staining and FITC-dextran intestinal paracellular permeability assay revealed that CORM-2 maintained the integrity of intestinal mucosal barrier and inhibited the changes of intestinal mucosal permeability. In addition, activation of T helper type 1 (Th1) and T helper type 17 (Th17) cells, and the increased expression levels of proinflammatory cytokines, such as interleukin-17 (IL-17) and interferon-gamma (IFN-γ), were observed in intestinal IRI process. In contrast, pretreatment with CORM-2 weakened changes of the abovementioned observations, in which inhibited activation of Th1 and Th17 cells. However, CORM-2 did not influence differentiation of regulatory T (Treg) cells in intestinal IRI progress. Notably, CORM-2 significantly upregulated the expression level of interleukin-10 (IL-10) protein and downregulated the expression levels of IL-17 and IFN-γ proteins in ileal tissues of rats. CORM-2 possessed anti-inflammatory effects in the progress of intestinal IRI by inhibiting activation of Th1 and Th17 cells in rats undergoing hemorrhagic shock.

Chemical Constituents of the Roots of Polygala tenuifolia and Their Anti-Inflammatory Effects

Increasing scientific evidence has demonstrated that the roots of Polygala tenuifolia Willd. have pharmacological effects related to anti-inflammation. Therefore, the aim of this study is to investigate the chemical constituents from P. tenuifolia roots as anti-inflammatory drug candidates. In the present work, twenty-three compounds were isolated from P. tenuifolia roots, including three saponins (1-3), ten phenylpropanoid sucrose esters (4-12), one benzoic acid sugar ester derivative (13), four xanthones (14-17), two hydroxy benzophenone derivatives (18 and 19), two phenolic derivatives (20 and 21), and two ionones (22 and 23). All isolates were tested for their inhibitory effects of LPS-stimulated NO and PGE2 production in RAW 264.7 macrophages. Among these, 3-O-(3,4,5-trimethoxy-cinnamoyl),6'-O-(p-methoxybenzoyl) sucrose ester (TCMB; 11) together with compounds 3 and 21 exhibited significant inhibitory effects on NO production, while TCMB and compounds 17, 19, and 21 showed strong inhibitory effects on PGE2 production. Specifically, TCMB (11) downregulated the protein levels of iNOS and COX-2 in LPS-induced RAW 264.7 macrophages. In addition, TCMB (11) dose-dependently diminished the relative mRNA expression levels of iNOS, PGE2, and proinflammatory cytokines (TNF-α, IL-1β, and IL-6). A molecular docking study showed that TCMB (11) has strong binding affinities with iNOS and COX-2.

Porphyromonas gingivalis infection promotes inflammation via inhibition of the AhR signalling pathway in periodontitis.

Porphyromonas gingivalis (P. gingivalis) is a key pathogen of chronic periodontitis. Aryl hydrocarbon receptor (AhR) is essential in immune homeostasis via modulation of pro-inflammatory cytokines production and indoleamine 2,3-dioxygenase (IDO). In this study, it is demonstrated that P. gingivalis may regulate AhR signalling in periodontitis, which provides a potential target for further immune regulation studies in periodontitis. Experimental periodontitis was induced in C57BL/6 mice by silk ligature and P. gingivalis oral inoculation. The alveolar bone resorption was examined using Micro-CT. Histological structures were observed and related cytokines involved in AhR signalling pathway were analysed. RAW264.7 cells were pretreated with AhR agonist (FICZ) and antagonist (CH223191) and infected with P. gingivalis subsequently. The levels of IDO, AhR and other related cytokines were measured. To demonstrate IDO activity, the concentrations of tryptophan (Trp) and kynurenine (Kyn) were assessed by HPLC. Histological analysis of periodontitis mice showed distinct alveolar bone resorption and inflammatory cell infiltration. The level of AhR and its downstream target factors were significantly decreased in inflamed gingival tissue. Furthermore, RAW 264.7 cells incubated by P. gingivalis exhibited increased pro-inflammatory cytokines production and decreased AhR, CYP1A1, CYP1B1, and IDO expression. Decreased IDO activity was observed as decreased Kyn/Trp ratio in the supernatant. Moreover, FICZ decreased the pro-inflammatory cytokines levels in P. gingivalis infected cells. It is concluded that P. gingivalis may promote inflammatory responses via inhibiting the AhR signalling pathway in periodontitis.

Alterations in the Humoral Immunophenotype of Children with Sickle Cell Disease on Hydroxyurea

Background: Sickle Cell Disease (SCD) is a red blood cell disorder associated with hemolysis and inflammation affecting homeostasis of the blood system. Both innate and T-cell driven adaptive immune systems are involved in SCD-linked inflammation. The involvement of B-cell compartment in etiology of SCD is much less studied. While complex interactions of ischemia-reperfusion, inflammation, and hemolysis affecting the bone marrow and immune system have been studied in adults with SCD, much less is known how their interplay affects SCD etiology in children. Objective: To determine the range of immune changes we assessed the frequency of 12 immune populations via flow cytometry, evaluated B cell related cytokines, and performed bulk RNA sequencing of the peripheral blood (PB) of pediatric patients with SCD as compared to pediatric race-matched controls as well adult patients with SCD as compared to adult race-matched controls. Method: Adult control PB was obtained from Rhode Island Blood Center. Pediatric control and pediatric and adult SCD PB were obtained from Rhode Island Hospital Hematology and Primary Care clinics. All informed consents/assents were obtained. PB samples were stained using markers identifying 12 immune populations including CD8, CD4, T Regulatory, B cells, Plasmablasts, Monocytes, Dendritic, Granulocytes, Neutrophils, Basophils, Eosinophils, and Stem Cells. Flow cytometry was performed on BDTM LSRII and analyzed using FlowJo. RNA was isolated from pediatric SCD or control PB and RNA-seq was performed by Novogene. Plasma from all patients (SCD and controls) was processed for cytokine analysis using Biolegend's LEGENDplex Human B Cell Panel. Demographics, clinical, and laboratory data were obtained from chart review. Results: Thirteen pediatric patients (ages 1-16, mean 8.7 years) and 12 adult patients (ages 8-15, mean 32.2 years) with SCD on Hydroxyurea were enrolled. HbSS genotype was found in 92.3% pediatrics and 66.7% adults. Mean Hydroxyurea dose was 23.4 mg/kg or 20.0 mg/kg for pediatric or adult SCD, respectively. Five pediatric controls (ages 9-14, mean 11.4 years) and 10 adult controls (ages 18-53, mean 31.0 years) were enrolled. Laboratory data indicated elevated hemolysis in pediatric and adult SCD with mean Absolute Reticulocyte Count (ARC) 203 ± 94.2 for pediatric SCD and 214 ± 169 for adult SCD. Flow cytometry data revealed 1.9-fold increase (p= 0.012) of CD3+CD4+ cells in Pediatric SCD versus pediatric controls, this change was not seen in adult SCD versus adult controls (Table 1). Pediatric SCD patients had 3.5-fold increase (p=0.005) in CD19+ B cells without further increase in differentiated B cells, this change was not seen in adult SCD versus adult controls (Table 1). Subsequent analysis of B cell related cytokines revealed significant increase in IL2 (p =0.013), IL4 (p=0.026), TNFβ (p= 0.039), and IL13 (p=0.034) in pediatric SCD versus pediatric controls; there was no significance for these cytokines in adult SCD versus adult controls. BAFF was elevated in both pediatric and adult SCD versus age-matched controls (p=0.041 and p=0.005, respectively). Finally, bulk RNA sequencing of pediatric SCD or control PB indicated significant changes in transcripts linked to humoral immunity. Over Representation Analysis (ORA) of cell type revealed upregulation of transcripts linked to pro-B cells (29 genes), plasma cells (37 genes) and CD34 Positive Pre-B cells (10 genes), (Figure 1). These results suggested significant alteration of the B cell maturation in pediatric SCD samples. Conclusions: Our data indicate significant changes in CD4+ and CD19+ cells in pediatric SCD patients versus controls. Increased CD4+ cell frequencies in children with SCD were previously reported (PMID 29926339). Our new finding of altered humoral immunity and differentiation of B cells noted by flow cytometry were consistent with elevated B-cell related cytokine levels in pediatric SCD versus pediatric controls. Elevated levels of pro-B cells, plasma cells, and CD34+ pre-B cell transcripts detected by RNA-seq in pediatric SCD patients versus controls further confirmed humoral alterations in pediatric SCD samples. Our findings point to previously unreported effect of SCD on the humoral immune system in children with SCD treated with Hydroxyurea, warranting further investigation. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Evaluation of Anti-Asthmatic Activity of Essential Oils from the Lauraceae Family in Lipopolysaccharide (LPS)-Stimulated NCI-H292 Cells

The Lauraceae family has commercial uses, such as in the food, pharmaceutical, and perfume industries. This study was conducted to investigate anti-asthmatic activity of essential oils from the seven species in the Lauraceae family. The essential oils were extracted from the leaves of seven species, and the chemical composition was investigated by gas chromatography-mass spectrometry. The major constituents of essential oils differed depending on the species, even if they belonged to the same family. The main constituents were camphor (89.09%) in Cinnamomum camphora, linalool (26.91%) in Cinnamomum cassia, 1,8-cineole (23.90%) in Cinnamomum japonicum, d-limonene (10.27%) and β-eudesmol (10.03%) in Lindera obtusiloba, δ-cadinene (13.85%) and α-phellandrene (11.57%) in Machilus japonica, cis-,trans-β-ocimene (13.80% and 12.06%) and elemol (11.46%) in Neolitsea aciculata, and cis-β-ocimene (37.94%) and sabinene (24.91%) in Neolitsea sericea. The anti-asthmatic activity of essential oils was investigated using the lipopolysaccharide-induced NCI-H292 cells. The relative expression levels of the pro-inflammatory cytokines [interleukin (IL)-1β and IL-6] and mucus gene (MUC5AC and MUC5B) were significantly reduced by essential oils from seven species in the Lauraceae family. Among the seven essential oils, the essential oil from L. obtusiloba had the most superior anti-asthmatic activity. These results suggest that the essential oil of L. obtusiloba leaves could be used as an agent to suppress mucus hypersecretion.

Inhibition of TNBS-induced intestinal inflammation in crucian carp (Carassius carassius) by oral administration of bioactive Bioactive food derived peptides

Intestinal enteritis is a main issue in crucian carp production which results in massive economic loss. Traditional antibiotics used for disease prevention of crucian carp (Carassius carassius) have been banned, thus an alternative approach needs to be identified. In this study, the bioactive peptide was evaluated as a diet supplement for preventing intestinal inflammation in crucian carp. Intestinal inflammation was induced by intrarectal administration of a 2,4,6-trinitrobenzene sulfonic acid (TNBS) solution. The fish samples were fed with different diets for 14 days. The disease activity index (DAI), which included, fish swimming, food intake, anal inflammation, body surface, and ascites was determined daily. Intestine segments were stained with haematoxylin and eosin (H.E.) for histopathological analysis. The expression of cytokines, including interleukin-1β (IL-1β), interleukin-8 (IL-8), tumor necrosis factor α (TNF-α), and myeloperoxidase (MPO) in crucian carp were determined. In TNBS-induced groups, the DAI scores were dramatically increased compared to the control group. The histopathological analysis showed that the damage of the fish intestine after the injection of TNBS. The relative expression levels of pro-inflammation cytokines (TNF-α, IL-1β, IL-8, MPO) were significantly increased compared to the control group on day 1. In the TNBS-induced group feed with a diet supplemented with bioactive peptide, the symptoms of intestinal inflammation were relieved on day 3 and the mRNA expression levels of pro-inflammation cytokines (TNF-α, IL-1β, IL-8, MPO) were reduced compared to day 1. On day 7, the fish samples enrofloxacin group and bioactive peptide group were recovered from TNBS-induced intestinal inflammation. This study showed that the fish diet supplemented with bioactive peptide could help to prevent and recover from intestinal inflammation. Thus, the bioactive peptide can be used as a replacement for antibiotics to prevent disease in aquaculture production.

Pro- and anti-inflammatory cytokine profiles in Plasmodium Falciparum-infected individuals from Baiyeku, Ikorodu, Lagos, Nigeria

Available evidence indicates that the various stages of the malaria parasite life cycle elicit specific immune responses of which the relative levels of pro-inflammatory cytokines are key to disease progression, killing the parasite and mediating disease outcomes. This study investigated T-cell response in malaria. Four hundred and sixty-two participants were screened in a community survey of Plasmodium falciparum malaria in Baiyeku, Lagos, Nigeria. P. falciparum parasitaemia was determined by microscopy while the serum levels of IL-10, IFNγ and TNFα were determined by Enzyme-linked immunosorbent assay (ELISA). A total of 70 (15.2 %) participants were microscopically positive for P. falciparum of which 70% were females, 30% were males while children aged 1-17 years were 65.7%. The geometric mean parasite density (GMPD) was significantly (p=0.001) higher among females than males. The GMPD of participants &lt;5 years of age was also significantly (p=0.001) higher than other age groups. About 46.8% of the participants were underweight (Body Mass Index, BMI &lt; 18.5) and also had the highest parasite intensity. The TNFα, IFNγ and IL-10 levels were significantly (p&lt; 0.05) higher in the infected than the uninfected participants. IFN-γ values were significantly (p=0.014) elevated among the symptomatic than the asymptomatic participants while there was no significant difference (P&gt;0.053) in the levels of TNF-α and IL-10 (P&gt;0.093) between the symptomatic and asymptomatic participants. The prevalence of P. falciparum obtained in this study area which is endemic to malaria is 15.2% suggesting a significant reduction of the disease over time due to awareness of the disease in the community. The levels of pro-inflammatory cytokines (IFN-γ and TNF-α) in this study were lower due to the down-regulatory action of the anti-inflammatory cytokines (IL-10). These findings suggest that higher levels of anti-inflammatory cytokines, especially IL-10 levels may contribute to the pathogenesis of uncomplicated malaria.