IL-6 Concentrations Research Articles

Cholesterol to saturated fat index (CSI), metabolic parameters and inflammatory factors among obese individuals

BackgroundThe role of dietary fat quality in promotion of cardiovascular diseases is studies before. However, the results are inconsistent. Recently, cholesterol to saturated fatty acid index (CSI) is suggested as a novel indicator of the atherogenicity and thrombogenicity potential of a diet. However, due to limited number of studies, in the current cross-sectional study, we aimed to evaluate the role of CSI in metabolic and inflammatory response among obese individuals.MethodsIn the current cross-sectional study 488 obese individuals aged 18–50 years old were involved in volunteer based invitation from outpatient obesity clinics. Subjects underwent anthropometric assays including weight, height, waist circumference (WC) and body composition and their fasting blood sample were obtained for biochemical assessments including blood sugar, serum lipids, hs-CRP and IL-6 concentrations by commercial kits. Physical activity was also assessed by short form of international physical activity questionnaire (IPAQ).ResultsAccording to our results, being at the top tetile of CSI was associated with higher anthropometric indices including weight, height, WC, FFM, and basal metabolic rate (BMR) compared with those at the lowest tertile (P < 0.05). Similarly, those at the highest category of CSI had significantly higher levels of serum glucose and hs-CRP both in crude and adjusted models in ANCOVA and in multinomial logistic regression models (P < 0.05).ConclusionIn the current study, for the first time, we identified the possible triggering role of dietary cholesterol to saturated fat index in increasing serum glucose and hs-CRP levels. due to cross-sectional design of the current study, causal inference is impossible. Further studies will help for better scientific justification.

NLRC5 exerts anti-endometriosis effects through inhibiting ERβ-mediated inflammatory response

BackgroundEndometriosis is well known as a chronic inflammatory disease. The development of endometriosis is heavily influenced by the estrogen receptor β (ERβ), while NOD-like receptors (NLRs) family CARD domain-containing 5 (NLRC5) exhibits anti-inflammatory properties during endometriosis. However, whether NLRC5-mediated anti-inflammation is involved in the ERβ-mediated endometriosis is still uncertain. This study aimed to assess that relation.MethodsNine cases of eutopic endometrial tissue and ten cases of ectopic endometrial tissue were collected from patients with endometriosis, and endometrial samples from ten healthy fertile women were analyzed, and the expression levels of ERβ were quantified using immunohistochemistry (IHC). Subsequently, we constructed mouse model of endometriosis by intraperitoneal injection. We detected the expression of ERβ, NLRC5, tumor necrosis factor-alpha (TNF-α), interleukin (IL)-6, and IL-10 and measured the volume of ectopic lesions in mice with endometriosis. In vitro, human endometrial stromal cells (hESCs) were transfected respectively with ERβ-overexpressing and NLRC5-overexpressing plasmids. We then assessed the expression of ERβ and NLRC5 using quantitative real-time PCR (qRT-PCR) and western blot analysis. Furthermore, we measured the concentrations of TNF-α, IL-6, and IL-10 in the cell culture supernatant through enzyme-linked immunosorbent assay (ELISA). Additionally, we evaluated the migration and invasion ability of hESCs using transwell and wound healing assays.ResultsInhibition of NLRC5 expression promotes the development of ectopic lesions in mice with endometriosis, upregulates the expression of pro-inflammatory factors TNF-α and IL-6, and downregulates the expression of anti-inflammatory factor IL-10. The high expression of NLRC5 in endometriosis depended on the ERβ overexpression. And ERβ promoted the migration of hESCs partially depend on inflammatory microenvironment. Lastly, NLRC5 overexpression inhibited ERβ-mediated development and inflammatory response of endometriosis.ConclusionsOur results suggest that the innate immune molecule NLRC5-mediated anti-inflammation participates in ERβ-mediated endometriosis development, and partly clarifies the pathological mechanism of endometriosis, expanding our knowledge of the specific molecules related to the inflammatory response involved in endometriosis and potentially providing a new therapeutic target for endometriosis.

Bioinformatics and Chemoinformatics Analysis Explored the Role of Linum usitatissimum in Diabetic Heart Conditions: Experimental Analysis in H9c2 Rat Embryonic Cardiomyocytes Cell Lines.

Cytokine storms and inflammation lead to heart failure (HF). Bioactive compounds, as complementary medicine, can be the primary source of compounds with anti-inflammatory properties. Linum usitatissimum (LiU) has antioxidant capacity and anti-inflammatory activity. Here, candidate hugeness was selected based on the in silico studies, bio-cheminformatics, and bioinformatic analysis for excremental validation. We selected the vital genes with differential expression from the GSE26887 dataset. Based on the bioinformatics analysis, several parameters are determined to choose switchable genes involved in diabetic HF (DHF). We designed the protein-protein interactions network to consider the nodes' degree, modularity, and betweenness centrality. Hence, we selected the interleukin (IL)-6 protein as a target for drug design and discovery to reduce diabetes complications in the heart. Here, H9c2 cell lines of rat embryonic cardiomyocytes induce HF using hyperglycemic and hyperlipidemic conditions. Real-time polymerase chain reaction evaluated the relative expression of SMAD7/NRF-2/STAT3. Furthermore, we assessed the concentration of IL-6 using the enzyme-linked immunosorbent assay technique. Based on the bioinformatic analysis, we found that IL-6 with the highest network parameters score might be presented as a druggable protein in the DHF. Bioactive compounds and phytochemicals have potential strategies to manage DHF. LiUs decreased the expression level of the SMAD7 (P <0.0001) and STAT3 (P < 0.0001), and increased the expression level of the NRF2 (P < 0.0001). In addition, LiUs significantly reduced the concentration of IL-6 (P < 0.0001). Our data proposed that LiUs regulated inflammation and triggered the antioxidant defense in HF. Moreover, LiUs could have potential approaches to managing and preventing DHF.

Abstract P390: The Adipokine Chemerin Does Not Promote Activation or Cytokine Secretion by T Cells Stimulated With a Strong Polyclonal Activator

High fat diet has long been implicated as a risk factor for obesity and other conditions associated with metabolic syndrome, including hypertension. The mechanism by which high fat diet increases risk of hypertension is likely multifactorial, but corresponding low-grade inflammation may play an important role. Low-grade inflammation associated with high fat diet is often accompanied by altered adipokine levels, including an increase in circulating chemerin. However, our understanding of the effects of chemerin and other adipokines on immune cells is limited. Thus, the purpose of these studies was to test the hypothesis that the adipokine chemerin promotes CD4 and CD8 T cell activation and cytokine secretion. To test this hypothesis, primary mouse splenocytes were treated with chemerin or vehicle 30 min prior to activation with a T cell-specific activator (anti-CD3/anti-CD28) for 24 h. T cell activation causes a rapid upregulation of cell surface proteins that are commonly used as experimental markers of activation. We found that chemerin did not impact the induction of the T cell activation markers, CD25, CD69, CD44, CD134, CD137 and CD154, in either CD4 or CD8 T cells. Likewise, we did not see a difference in the downregulation of CD62L, which is rapidly internalized by T cells following activation. Consistent with this, chemerin caused no difference in cytokine secretion by activated T cells. Specifically, the concentrations of IL-2, IL-17A, IFNγ, and TNFα were similar between groups. Overall, this study shows that the adipokine chemerin does not promote acute changes in activation or cytokine expression in primary mouse splenic T cells fully activated with anti-CD3/anti-CD28.

Relationship of Inflammatory Mediators (Interleukin and Cortisol Concentrations) with Corneal Epithelial Quantifiable Metrics

PurposeTo investigate the relationship of inflammatory biomarkers with corneal epithelial quantifiable metrics in patients with keratoconus and in healthy eyes. DesignProspective observational comparative study. ParticipantsThis study included 100 eyes of 100 patients, 48 eyes of 48 patients with KC, and 52 healthy eyes of 52 healthy controls. MethodsThe concentrations of tear cytokines were investigated in both groups: interleukin (IL) 1B, IL6, IL8, IL10, IL12p70 and TNFα were obtained by capillary flow and measured using flow cytometer. Cortisol concentrations were determined in both groups from the most proximal hair segment as an index of cumulative secretion and measured by liquid chromatography mass spectrometry. Epithelial variables were obtained with optical coherence tomography (OCT). Pearson correlation (r) was used to measure linear dependence between two different variables. Main Outcome MeasuresInvestigating the existence, strength, and significance of any correlation between inflammatory biomarkers (IL1B, IL6, IL8, IL10, IL12p70, TNFα and hair cortisol concentration) and OCT corneal epithelial quantifiable variables such as minimum and maximum epithelial thickness of the map, difference between the minimum and maximum (Epithelial Min-Max) and standard deviation of the epithelial thickness of the map (Epithelial Std Dev) and average epithelial thickness of the superior and inferior regions of the map. ResultsEyes with keratoconus presented statistically significantly higher levels of IL1b (p=0.02), IL6 (p <0.0001), IL8 (p <0.0001), and TNFα (p <0.0001) and hair cortisol concentration (p=0.01) compared to healthy controls.There was a significant correlation between IL6 and measurement Epithelium Min-Max [Pearson= -0.59 (-0.69, -0.47); p<0.0001] and Epithelial standard deviation (Std Dev) [Pearson= +0.56 (0.44, 0.67); p<0.0001]. There was a significant correlation between hair cortisol concentration and Epithelium Min-Max [Pearson=-0.27 (-0.42, -0.1); p<0.0001] and Epithelium Std Dev groups [Pearson= +0.2 (0.03, 0.36); p=0.021]. There was also a significant correlation between TNFα and Epithelial Max. [Pearson= -0.37 (-0.55, 0.17); p<0.0001]. We found no significant correlation between the concentration of IL1b, IL8, IL10 and IL2p70 with any epithelium parameters. ConclusionsThe higher concentration of inflammatory markers (IL6 and hair cortisol) in eyes with keratoconus present a significant correlation with OCT metrics identifying epithelial variability, such as Epithelial Min-Max and Std Dev. These findings demonstrate the role of chronic inflammation in eyes with keratoconus, and that these epithelial changes detectable with OCT are sensitive to this inflammatory process.

Increased nuclear receptor subfamily 2, group E, member 1 (NR2E1) concentrations of PBMCs are associated with chronic inflammation in overweight/obesity

BackgroundChronic inflammation plays a crucial role in the pathogenesis of overweight/obesity. Nuclear receptor subfamily 2, group E, member 1 (NR2E1) is one of the nuclear receptor family proteins that play crucial roles in regulating numerous life processes. In this study, we attempted to detect NR2E1 levels in peripheral blood mononuclear cells (PBMCs) of overweight/obese people and preliminarily elucidate the regulatory role of NR2E1 in obesity-related chronic inflammation. MethodsWe conducted a cross-sectional analysis of the clinical and biochemical data from 62 overweight/obese people and 70 control subjects. PBMCs of the participants were collected for detection of NR2E1 levels. PBMCs isolated from the control subjects were treated with different concentrations of palmitic acid (PA). We also transfected p-EGFP-N1-NR2E1 plasmids into PBMCs and treated them with PA, then detected TNF-α and IL-6 concentrations in the supernatant of PBMCs. ResultsThe NR2E1 mRNA and protein levels in overweight/obese people were both significantly higher than those in normal-BMI people (p < 0.01). NR2E1 mRNA levels in PBMCs of overweight/obese people were positively related with TC, FFA, IL-6, TNF-α (r = 0.387, 0.440, 0.610, 0.530, p < 0.01) and LDL-c (r = 0.290, p < 0.05). A similar correlation was also found between NR2E1 protein levels and these parameters. The expression of NR2E1 in PBMCs from the control subjects increased apparently with the treatment of PA in a concentration-depend manner in vitro. Overexpression of NR2E1 in PBMCs decreased TNF-α and IL-6 expression induced by PA (p < 0.01). ConclusionNR2E1 levels are increased in overweight/obese people and have a positive relationship with TC, FFA, LDL-C, TNF-α and IL-6. Overexpression of NR2E1 could alleviate PA-induced chronic inflammation. NR2E1 may be a potential target for regulating chronic inflammation in obesity.

Evaluation of Regulatory B Cells and Serum IL-10 Concentration in Peripheral Blood of Women with Recurrent Pregnancy Loss

Background: Although the role of B cells in normal pregnancy has been recently highlighted, their importance and function are not completely clarified. Until now, some investigations have shown that during pregnancy, regulatory B cells (Breg), a subset of B cells, are one of the key players in immune regulation by both producing IL-10 and cell-cell interactions. Therefore, any decrease in the number or function of these cells may lead to recurrent pregnancy loss (RPL). Thus, the objective of this study was to characterize Breg cell frequency and function in women who suffered from RPL in comparison with healthy non-pregnant and pregnant women (under twenty weeks of gestational age) as controls. Method: In this study, peripheral blood samples of women suffering from RPL (n=8), women with normal pregnancy under 20 weeks of gestational age (n=14), and healthy nonpregnant women (n=10) were collected. The frequency of Breg cells (CD19+CD24hiCD38hi) was measured by flow cytometry. The serum level of the IL-10 cytokine, as a marker of Breg cell function, was measured by ELISA. Results: The Percentage of Breg cells in women who suffered from RPL was significantly lower than that of women who had normal pregnancies (P=0.0016). The percentages of Breg cells in women who suffered from RPL were also significantly lower than in non-pregnant women (P=0.0001). Furthermore, no significant differences were observed in Breg cell percentages between normal pregnant and non-pregnant women. Evaluation of IL-10 concentration in the serum of women who had participated in this study showed no significant differences between the three groups. Conclusion: Based on our results, the number of Breg cells was significantly lower in RPL women than in healthy non-pregnant and normal-pregnant women, which shows the significance of these cells in the maintenance of normal pregnancy. However, we could not detect significant differences in the serum levels of IL-10, bringing to mind the notion that the beneficial and supportive function of these cells during pregnancy might be independent of IL-10 secretion. by these cells. Thus, screening of Breg cells in women with pregnancy complications, especially RPL, could be helpful for predicting a healthy pregnancy.

Oral mucositis management with photobiomodulation, Bidens pilosa L. (Asteraceae) and Curcuma longa L. (Zingiberaceae), the FITOPROT herbal medicine, and its influence on inflammatory cytokine levels: a randomized clinical trial.

This randomized clinical trial aimed to compare the effects of a mucoadhesive formula, containing curcuminoids from Curcuma longa L. and glycerinated extract of Bidens pilosa L. (FITOPROT), associated with photobiomodulation (PBM), and of PBM exclusively, on the incidence of oral mucositis (OM)-induced by radiotherapy (RT) in the head and neck region, and the salivary expression of inflammatory cytokines, in patients with head neck cancer. Patients were randomly assigned into two intervention groups-FITOPROT + PBM (n = 25) or PBM (n = 27). PBM protocol comprised a wavelength of 660nm, 25 mW, 0.25J/point, and daily irradiation from the first until the last day of RT. FITOPROT was gargled twice a day. All patients underwent a preventive oral care program throughout the study. OM degree, salivary concentration of nitrite, and inflammatory (IL-1, TNFα, IL-6, IL-8, and IL-12p70), and anti-inflammatory (IL-10) cytokines were assessed at baseline, and at the 7th, 14th, 21st, and 30th RT sessions. There were no differences in the OM degree between groups, but the RT dose significantly affected the OM. The RT significantly affected the salivary nitrite, TNFα, IL-1β, and IL-10 concentrations. FITOPROT associated with PBM showed limited effects on preventing the incidence of severe OM compared to PBM alone. However, FITOPROT + PBM may be associated with nitrite and cytokine balance, which may contribute to the occurrence of fewer cases of severe OM. Brazilian Clinical Trials database (ReBEC; RBR-9vddmr), registered UTN code: U1111-1193-2066, registered in August 8th, 2017.

Exosomes secreted by podocytes regulate the differentiation of Th17/Treg cells in idiopathic nephrotic syndrome

BackgroundPrevious studies have demonstrated that immune cells release exosomes, which act as antigen-presenting vesicles to activate T cells. In our previous study, we discovered that podocytes, a type of kidney cell, can also exhibit antigen-presenting functions to naïve CD4+ T cells in idiopathic nephrotic syndrome (INS). Building upon these findings, the objective of this study was to investigate whether podocytes can regulate the balance between Th17 and Treg cells through the release of exosomes. MethodsWe co-cultured naïve CD4+ T cells with LPS-treated bone marrow dendritic cells (LPS-BMDC), LPS-treated mouse podocyte clone 5 (LPS-MPC-5), and exosomes derived from LPS-MPC-5 (LPS-EXO). As a control group, naïve CD4+ T cells were cultured with exosomes from untreated MPC-5 (EXO). After 48 h, we analyzed the percentages of Th17 and Treg cells using flow cytometry, measured the concentrations of IL-17A, IL-10, and IL-4 were using ELISA, and examined the expressions of IL-17a, IL-10, RORC, and FOXP3 using RT-qPCR. ResultsWe confirmed the presence of exosomes derived from podocytes based on their morphology, size distribution, concentrations, and the levels of exosomes-specific markers. The percentage of Th17 and Treg cells in the LPS-EXO group was significantly higher than that in the control groups, but lower than in the LPS-MPC-5 group. Furthermore, the ratio of Th17/Treg was relatively higher in the LPS-EXO group compared to the LPS-MPC-5 group. ConclusionThis study indicated further insights into the role of exosomes released from LPS-treated podocytes in regulating the balance between Th17 and Treg cells in INS.

Gut microbiota and inflammation analyses reveal the protective effect of medium-chain triglycerides combined with docosahexaenoic acid on cognitive function in APP/PS1 and SAMP8 mice

Accumulating evidence has demonstrated that medium-chain triglycerides (MCTs) and docosahexaenoic acid (DHA) positively affect cognitive function. However, it remains unclear whether the improvement is related to the alterations of gut microbiota and inflammation and the impact of the combined intervention. In this study, we hypothesized that the supplementation of MCTs combined with DHA could modulate gut microbiota, inflammation, and improve cognitive function in APPswe/PS1De9 (APP/PS1) model mice and senescence-accelerated mouse prone-8 (SAMP8), which are two different mouse models used in neurodegeneration research. The mice were divided into four groups: Control group, MCTs group, DHA group and MCTs+DHA group. The study assessed cognitive function, inflammatory cytokines, and gut microbiota composition. The results showed that supplementation of MCTs+DHA improved spatial learning ability, memory capacity, exploratory behavior; decreased the relative abundance of Proteobacteria; reduced the ratio of Firmicutes/Bacteroidetes (F/B); decreased the concentrations of serum interleukin (IL)-2, IL-6, monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-alpha), while increasing the concentration of IL-10. Furthermore, supplementation with MCTs+DHA exhibited significantly superior effects compared to MCTs or DHA alone in reducing inflammation, optimizing gut microbiota composition, and improving cognitive function. In conclusion, supplementation with MCTs+DHA improved cognition function, accompanied with favorable alterations in gut microbiota and inflammation in APP/PS1 and SAMP8 mice.

The role of cytokines in acute and chronic postsurgical pain after major musculoskeletal surgeries in a quaternary pediatric center

Study ObjectiveTo determine if baseline cytokines/chemokines and their changes over postoperative days 0–2 (POD0-2) predict acute and chronic postsurgical pain (CPSP) after major surgery. DesignProspective, observational, longitudinal nested study. SettingUniversity-affiliated quaternary children’s hospital. PatientsSubjects (≥8 years old) with idiopathic scoliosis undergoing spine fusion or pectus excavatum undergoing Nuss procedure. MeasurementsDemographics, surgical, psychosocial measures, pain scores, and opioid use over POD0-2 were collected. Cytokine concentrations were analyzed in serial blood samples collected before and up to two weeks after surgery, using Luminex bead arrays. After data preparation, relationships between pre- and post-surgical cytokine concentrations with acute (% time in moderate-severe pain over POD0-2) and chronic (pain score > 3/10 beyond 3 months post-surgery) post-surgical pain were analyzed using univariable and multivariable regression analyses with adjustment for covariates and mixed effects models were used to associate longitudinal cytokine concentrations with pain outcomes. Main ResultsAnalyses included 3,164 repeated measures of 16 cytokines/chemokines from 112 subjects (median age 15.3, IQR 13.5–17.0, 54.5 % female, 59.8 % pectus). Acute postsurgical pain was associated with higher baseline concentrations of GM-CSF (β = 0.95, SE 0.31; p = 0.003), IL-1β (β = 0.84, SE 0.36; p = 0.02), IL-2 (β = 0.78, SE 0.34; p = 0.03), and IL-12 p70 (β = 0.88, SE 0.40; p = 0.03) and longitudinal postoperative elevations in GM-CSF (β = 1.38, SE 0.57; p = 0.03), IFNγ (β = 1.36, SE 0.6; p = 0.03), IL-1β (β = 1.25, SE 0.59; p = 0.03), IL-7 (β = 1.65, SE 0.7; p = 0.02), and IL-12 p70 (β = 1.17, SE 0.58; p = 0.04). In contrast, CPSP was associated with lower baseline concentration of IL-8 (β = -0.39, SE 0.17; p = 0.02), and the risk of developing CPSP was elevated in patients with lower longitudinal postoperative concentrations of IL-6 (β = -0.57, SE 0.26; p = 0.03), IL-8 (β = -0.68, SE 0.24; p = 0.006), and IL-13 (β = -0.48, SE 0.22; p = 0.03). Covariates female (vs. male) sex and surgery type (pectus surgery vs. spine) were associated with higher odds for CPSP in baseline adjusted cytokine-CPSP association models for IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, TNFα, and IL-8, IL-10, respectively. ConclusionWe identified pro-inflammatory cytokine profiles associated with higher risk of acute postoperative pain. Interestingly, pleiotropic cytokine IL-6, chemokine IL-8 (which promotes neutrophil infiltration and monocyte differentiation), and monocyte-released anti-inflammatory cytokine IL-13, were associated with lower CPSP risk. Our results suggest heterogenous outcomes of cytokine/chemokine signaling that can both promote and protect against post-surgical pain. These may serve as predictive and prognostic biomarkers of pain outcomes following surgery.

LOCAL IMMUNE STATUS OF PATIENTS AFTER HYSTEROSCOPIC MYOMECTOMY AND METHODS OF ITS CORRECTION

Aim. To evaluate changes in the local immune status of women after hysteroscopic myomectomy and the effect of the proposed treatment complex on them. Materials and methods. 221 patients with submucous uterine myoma who underwent hysteroscopic myomectomy were examined. 121 patients of the main group in the semi-operative period received complex rehabilitation therapy, 100 patients of the comparison group did not receive additional treatment after surgery. 80 women who applied for a routine preventive examination formed the control group. In all patients, the content of factors of local immunity – IL-6, IL-10, lactoferrin and secretory inhibitor of leukocyte proteinase in cervical mucus was studied by enzyme-linked immunosorbent assay (ELISA) using Atiscare reagents in the laboratory conditions of the Center. The indicators were studied before surgery, 3 and 6 months after surgery. Results. In patients with submucosal uterine myoma, an increased content of pro-inflammatory IL-6 in cervical mucus was found (357.9 pg/ml and 345.3 pg/ml), in the control group 134.7 pg/ml and a decrease in the concentration of anti-inflammatory IL-10 (37 .6 pg/ml and 34.3 pg/ml) while the value of the control group was 78.5 pg/ml. The increase in the cervical content of the secretory inhibitor of leukocyte proteinase in women with submucous uterine myoma (274.2 pg/ml and 284.9 pg/ml against 114.5 pg/ml in the control group) is evidence of an active reaction of the mucous membrane to the inflammatory process. The role of the factor in the implementation of antisperm immunity is a factor of impaired fertility in such women. Elevated lactoferrin (27.4 ng/ml and 28.4 ng/ml vs. 3.5 ng/ml in healthy women) is also associated with infertility. Performing hysteroscopic myomectomy leads to a gradual, but incomplete normalization of the content of the specified immune factors in the cervical mucus, and the improvement of the treatment complex leads to the full achievement of the indicator of the control group after 6 months, which is important for the implementation of reproductive function. Conclusions. 1. Patients with submucous uterine myoma are characterized by an increased content of pro-inflammatory factors of local immunity – IL-6, lactoferrin and secretory inhibitor of leukocyte proteinase, as well as a reduced concentration of anti-inflammatory IL-10. 2. After hysteroscopic myomectomy for 6 months, there is a decrease in the content of pro-inflammatory factors and an increase in the concentration of IL-10 in the cervical mucus, but the indicators do not reach the values of healthy women. 3. The use of the proposed complex of treatment after hysteroscopic myomectomy allows to achieve faster changes in the indicators of local cervical immunity, and after 6 months to achieve the values obtained in healthy women.

Interleukin-6, -17, and -35 levels in association with clinical status in stage III and stage IV periodontitis: a cross-sectional study

BackgroundThis study compared the concentrations of interleukin (IL)-6, IL-17, and IL-35 in the gingival crevicular fluid of periodontally healthy participants with individuals who had stage III and IV periodontitis.MethodsIn total, 60 participants with stage III grade B-C (n = 12)—stage IV grade C (n = 18) periodontitis and 30 healthy controls were included in this cross-sectional study. Full-mouth clinical periodontal measurements were performed. Concentrations of IL-6, IL-17, and IL-35 were determined using enzyme-linked immunosorbent assays. Parametric/nonparametric methods, Pearson’s/Spearman’s correlation, and logistic regression methods were used for data analyses.ResultsThe periodontitis group exhibited significantly higher levels of IL-6, IL-17, and IL-35 compared with the healthy group (p < 0.001). IL-17 levels had a positive correlation with pocket depth (PD) (r = 0.395; p = 0.031) in the periodontitis group. IL-6, IL-17, and IL-35 levels were associated with periodontitis (odds ratio [OR] = 1.344, 95% confidence interval [CI] = 1.159–1.56; OR = 1.063, 95% CI = 1.025–1.102; OR = 1.261, 95% CI = 1.110–1.434, respectively) (p < 0.001, p = 0.001, p < 0.001, respectively). Full-mouth and sampling sites PD and clinical attachment loss (CAL) values were significantly higher in the periodontitis group than in the healthy group (p < 0.001).ConclusionsThis study revealed upregulated levels of IL-6, IL-17, and IL-35 in periodontitis patients compared to healthy individuals. IL-17 shows a correlation with increased PD. These findings suggest a potential association between these cytokines and severe and advanced periodontitis.Trial registrationThe trial was registered in ClinicalTrials.gov with this identifier NCT05306860 on 24/01/2022.

РОЛЬ ЦИТОКИНОВ В ПРОГНОЗИРОВАНИИ ТЯЖЕСТИ СИСТЕМНОЙ ВОСПАЛИТЕЛЬНОЙ РЕАКЦИИ У ПАЦИЕНТОВ С СОЧЕТАННОЙ ТРАВМОЙ

In connection with changes in the level of blood cytokines and the development of post-traumatic complications, monitoring these indicators can help in understanding the role of blood cytokines in the development of inflammatory reactions after injury and in the future the possibility of using them as biomarkers to predict the development of the severity of the systemic inflammatory response. Aim. to study the role of cytokines in the development of inflammatory reactions after injury and the possibility of using them as biomarkers. Material and methods. The study included 228 patients with combined trauma of several organ cavities and skeletal injuries. Determination of the concentrations of IL-8, IL-10, TGFβ and IL-4 was carried out using the enzyme immunoassay method in strict accordance with the manufacturer's instructions (R &amp; D Systems, USA). Results As a result of the study, IL-6 increased in patients 1-2 days after injury, as did IL-8, IL-10, TGFβ and IL-4. The levels of IL-6, IL-8, IL-10, TGFβ in patients on days 5-7 remained high and tended to increase, on the contrary, the level of IL-4 tended to decrease. In our study, we detected an increase in TGFβ-1 on days 1-2 after the patient received injury and reached maximum values (TGFβ-1 -134 ng/ml). Conclusion. Changes in the levels of the studied blood cytokines in patients with injuries revealed that there is an imbalance in inflammatory reactions after injury, which was more pronounced in patients in severe conditions and in patients with complications.

CSF Cytokines and Chemokines Involved in Cytotoxic Cell Function and Risk of Acute 14-Day Mortality in Persons with Advanced HIV and Cryptococcal Meningitis.

Despite availability of HIV treatment globally, cryptococcal meningitis continues to cause considerable morbidity and mortality. The role of the immune response in acute mortality remains unclear. To investigate the immune environment in the central nervous system, cerebrospinal fluid (CSF) from 337 Ugandans with advanced HIV and first-episode cryptococcal meningitis was collected at time of hospitalization. Participants were treated with standard of care amphotericin-B and fluconazole. Cytokines and chemokines in the CSF were quantified and compared by 14-day survival, stratification by quartiles, and logistical regression to determine association with acute mortality. 84 (24.9%) of the participants died by day 14 of hospitalization. Persons who survived to day 14 had higher levels of proinflammatory macrophage inflammatory protein (MIP)-3β and interferon (IFN)-β and cytotoxicity-associated Granzyme-B and inflammatory protein (IP)-10 compared to those who died (P<.05 for each). Logistical regression analysis revealed that per two-fold increase in proinflammatory IL-6, IL-1α, MIP-1β, MIP-3β, and IFN-β and cytotoxicity-associated IL-12, TNF-α, Granzyme-B, and IP-10 CSF concentrations, the risk of acute 14-day mortality decreased. Similar biomarkers were implicated when stratified by quartiles and further identified that lower concentrations of anti-inflammatory IL-10 and IL-13 as associated with 14-day mortality (P<.05 for each). Proinflammatory and cytotoxicity-associated cytokine and chemokine responses in the CSF decrease the risk of acute 14-day mortality. These data suggest that a cytotoxic immune environment in the CSF could potentially improve acute survival. Further research on cytotoxic cells is crucial to improve understanding of innate and adaptive immune responses in cryptococcal meningitis.

Beetroot (Beta vulgaris L.) Extract Gives Superior Effect than Beetroot Juice on Increasing HDL and Decreasing LDL and IL-6 in Dyslipidemic Rats Model

BACKGROUND: Dyslipidemia derives from disturbances in lipid metabolism as an interaction between genetic and environmental factors, characterized by increased levels of cholesterol, triglycerides, low-density lipoprotein (LDL) and decreased levels of high-density lipoprotein (HDL). Continuous dyslipidemia will increase the risk of atherosclerosis. In dyslipidemia, macrophages release inflammatory cytokines such as interleukin (IL)-1, IL-6, IL-12, IL-15, IL-18, and then induce T-cell infiltration formed atherosclerotic plaques. Beet (Beta vulgaris L.) which contains active compounds and antioxidant, such as flavonoids and betacyanin, has the ability to reduce dyslipidemia. This study evaluated effect of beetroot juice and beetroot extract on dyslipidemic rat models by evaluating the level of HDL, LDL and IL-6.METHODS: Fourty-two male Sprague Dawley rats were divided into seven groups. Rats were fed either with standard food or high fat diet (HFD) for 28 days, then different interventions using either single combination of simvastatin, beetroot juice, or beetroot extract were given alongside the standard food or HFD diet for another 28 days. LDL and HDL were measured pre- and post-intervention, while IL-6 was measured only post-treatment.RESULTS: Daily administration of a single beetroot juice or beetroot extract or in combination with simvastatin reduced LDL significantly compared to pre-intervention. The interventions also increased HDL significantly and lower IL-6 concentrations compared to group that received no intervention.CONCLUSION: Administration of beetroot juice and beetroot extract can lower LDL, increase HDL, and decrease IL-6 either alone or in combination treatment with simvastatin, it is indicated beetroot have a potential benefit for prevention and therapeutic in dyslipidemia.KEYWORDS: beetroot juice, beetroot extract, LDL, HDL, IL-6

MiR-6721-5p as a natural regulator of Meta-VCL is upregulated in the serum of patients with coronary artery disease

BackgroundCoronary artery disease (CAD), the leading cause of mortality globally, arises from atherosclerotic blockage of the coronary arteries. Meta-vinculin (meta-VCL), a large spliced isoform of VCL, co-localizes in muscular adhesive structures and plays significant roles in cardiac physiology and pathophysiology. This study aimed to identify microRNAs (miRNAs) regulating meta-VCL expression and investigate the expression alterations of the miRNAs of interest and meta-VCL as potential biomarkers in the serum of CAD patients. MethodsBioinformatics tools were employed to select miRNAs targeting meta-VCL. Cell-based ectopic expression analysis and a dual-luciferase assay were used to examine the interactions between miRNAs and meta-VCL. An ELISA assessed the concentrations of interleukin-6 (IL-6), IL-10, and tumor necrosis factor-α (TNF-α). MiRNA and meta-VCL expression patterns and biomarker suitability were evaluated in serum samples from CAD and non-CAD individuals using real-time PCR. A cardiac cell-line data set and CAD blood exosome samples were analyzed using bioinformatics and ROC curve analyses, respectively. ResultsmiR-6721-5p directly interacted with the putative target sites at the 3′-UTR of meta-VCL and regulated its expression. IL-10 and TNF-α concentrations, which may act as anti-inflammatory factors, decreased following miR-6721-5p upregulation and meta-VCL downregulation. Bioinformatics and experimental expression analyses confirmed downregulated meta-VCL expression and upregulated miR-6721-5p expression in CAD samples. ROC curve analysis yielded an AUC score of 0.705 (P = 0.018), indicating the potential suitability of miR-6721-5p as a biomarker for CAD. ConclusionsmiR-6721-5p plays a regulatory role in meta-VCL expression and may contribute to CAD development by reducing anti-inflammatory factors. These findings suggest that miR-6721-5p could serve as a novel biomarker in the pathogenesis of CAD.

Immunological insights: assessing immune parameters in medical professionals exposed to SARS-CoV-2

BackgroundThe immunological background responsible for the severe course of COVID-19 and the immune factors that protect against SARS-CoV-2 infection are still unclear. The aim of this study was to investigate immune system status in persons with high exposure to SARS-CoV-2 infection.MethodsSeventy-one persons employed in the observation and infectious diseases unit were qualified for the study between November 2020 and October 2021. Symptomatic COVID-19 was diagnosed in 35 persons. Anti-SARS-CoV-2 antibodies were also found in 8 persons. Peripheral blood mononuclear cells subpopulations were analyzed by flow cytometry, and the concentrations of cytokines and anti-SARS-CoV-2 antibodies were determined by ELISA.ResultsThe percentages of cytotoxic T lymphocytes (CTLs), CD28+ and T helper (Th) cells with invariant T-cell receptors were significantly higher in persons with symptomatic COVID-19 than in those who did not develop COVID-19’ symptoms. Conversely, symptomatic COVID-19 persons had significantly lower percentages of: a) CTLs in the late stage of activation (CD8+/CD95+), b) NK cells, c) regulatory-like Th cells (CD4+/CTLA-4+), and d) Th17-like cells (CD4+/CD161+) compared to asymptomatic COVID-19’ persons. Additionally, persons with anti-SARS-CoV-2 antibodies had a significantly higher lymphocyte count and IL-6 concentration than persons without these antibodies.ConclusionNumerous lymphocyte populations are permanently altered by SARS-CoV-2 infection. High percentages of both populations: NK cells—as a part of the non-specific response, and T helper cells’ as those regulating the immune response, could protect against the acute COVID-19 symptoms development. Understanding the immune background of COVID-19 may improve the prevention of this disease by identifying people at risk of a severe course of infection.Trial registrationThis is a retrospective observational study without a trial registration number.

Uncovering the mechanisms of Zhubi decoction against rheumatoid arthritis through an integrated study of network pharmacology, metabolomics, and intestinal flora

Ethnopharmacological relevanceZhubi Decoction (ZBD) is a modified formulation derived from the classic traditional Chinese medicine prescription “Er-Xian Decoction” documented in the esteemed “Clinical Manual of Chinese Medical Prescription”. While the utilization of ZBD has exhibited promising clinical outcomes in treating rheumatoid arthritis (RA), the precise bioactive chemical constituents and the underlying mechanisms involved in its therapeutic efficacy remain to be comprehensively determined. Aim of the studyThis study aims to systematically examine ZBD's pharmacological effects and molecular mechanisms for RA alleviation. Materials and methodsUtilizing the collagen-induced arthritis (CIA) rat model, we comprehensively evaluated the anti-rheumatoid arthritis effects of ZBD in vivo through various indices, such as paw edema, arthritis index, ankle diameter, inflammatory cytokine levels, pathological conditions, and micro-CT analysis. The UPLC-MS/MS technique was utilized to analyze the compounds of ZBD. The potential therapeutic targets and signaling pathways of ZBD in the management of RA were predicted using network pharmacology. To analyze comprehensive metabolic profiles and identify underlying metabolic pathways, we conducted a serum-based widely targeted metabolomics analysis utilizing LC-MS technology. Key targets and predicted pathways were further validated using immunofluorescent staining, which integrated findings from serum metabolomics and network pharmacology analysis. Additionally, we analyzed the gut microbiota composition in rats employing 16 S rDNA sequencing and investigated the effects of ZBD on the microbiota of CIA rats through bioinformatics and statistical methods. ResultsZBD exhibited remarkable efficacy in alleviating RA symptoms in CIA rats without notable side effects. This included reduced paw redness and swelling, minimized joint damage, improved the histopathology of cartilage and synovium, mitigated the inflammatory state, and lowered serum concentrations of cytokines TNF-α, IL-1β and IL-6. Notably, the effectiveness of ZBD was comparable to MTX. Network pharmacology analysis revealed inflammation and immunity-related signaling pathways, such as PI3K/AKT, MAPK, IL-17, and TNF signaling pathways, as vital mediators in the effectual mechanisms of ZBD. Immunofluorescence analysis validated ZBD's ability to inhibit PI3K/AKT pathway proteins. Serum metabolomics studies revealed that ZBD modulates 170 differential metabolites, partially restored disrupted metabolic profiles in CIA rats. With a notable impact on amino acids and their metabolites, and lipids and lipid-like molecules. Integrated analysis of metabolomics and network pharmacology identified 6 pivotal metabolite pathways and 3 crucial targets: PTGS2, GSTP1, and ALDH2. Additionally, 16 S rDNA sequencing illuminated that ZBD mitigated gut microbiota dysbiosis in the CIA group, highlighting key genera such as Ligilactobacillus, Prevotella_9, unclassified_Bacilli, and unclassified_rumen_bacterium_JW32. Correlation analysis disclosed a significant link between 47 distinct metabolites and specific bacterial species. ConclusionZBD is a safe and efficacious TCM formulation, demonstrates efficacy in treating RA through its multi-component, multi-target, and multi-pathway mechanisms. The regulation of inflammation and immunity-related signaling pathways constitutes a crucial mechanism of ZBD's efficacy. Furthermore, ZBD modulates host metabolism and intestinal flora. The integrated analysis presents experimental evidence of ZBD for the management of RA.

Kai Yu Zhong Yu recipe mitigates stress-induced accelerated follicle loss in mice by regulating the interplay between apoptosis and autophagy via the SIRT1/FOXO1/3 pathway

BackgroundPsychological stress-related infertility is often attributed to a decline in both the quality and quantity of oocytes. Kai Yu Zhong Yu (KYZY) recipe, a classic herbal formula in Traditional Chinese Medicine (TCM), has been employed for centuries to address stress-related infertility. Our previous studies have demonstrated its effectiveness in counteracting the reduction in oocyte competence induced by psychological stress. PurposeTo investigate the mechanism by which KYZY mitigates the adverse effects of stress. MethodsFemale ICR mice were randomly assigned to one of five groups: control, CUMS, KYZY-H, KYZY-M, and KYZY-L. Except for the control group, the other groups underwent a six-week Chronic Unpredictable Mild Stress (CUMS) procedure. Following CUMS, the three KYZY groups were received high (38.2 g kg-1), medium (19.1 g kg-1), and low (9.6 g kg-1) doses of the KYZY recipe intragastrically for four weeks. Additionally, RNA interference was used to reduce Sirt1 expression in the ovary to assess the involvement of the SIRT1 pathway. ELISA, HE staining, follicle counting, TUNEL staining, and Western blot were conducted to explore the mechanism of KYZY in treatment of stress-related infertility. ResultsKYZY treatment significantly reduced concentrations of IL-6, IL-1β and TNF-α, reversed the loss of small follicles, and alleviated follicular apoptosis and autophagy. KYZY also elevated SIRT1/FOXO1/3 activity, reduced BAX expression, increased BCL-2 expression, and restored Beclin-1-dependent autophagy. Sirt1 silencing downregulated the activity of the SIRT1/FOXO1/3 pathway and triggered Caspase-3-mediated cleavage of Beclin-1 in mouse ovary. Conversely, Sirt1 silencing nullified the effect of KYZY recipe on inflammation and the regulation of apoptosis and autophagy. ConclusionKYZY enhances SIRT1/FOXO1/3 pathway activity, shifting the balance from apoptosis towards autophagy to safeguard oocyte capacity against the effects of psychological stress.