Interleukin-6 Protein Levels Research Articles

Lipocalin 2 inhibits the expressions of interleukin-8 and macrophage inflammatory protein-1α in human neutrophil-like cells.

Lipocalin 2 (LCN2) is a glycoprotein with multiple functions, including antimicrobial activity, inflammatory response modulation, and cell migration. LCN2 is expressed in some cells, such as epithelial cells and neutrophils, and its levels are increased in inflammatory diseases. This study investigated the presence of LCN2 receptor (24p3R) in cells around periodontal tissues and function of LCN2 in cells with a receptor to explore the role of LCN2 in periodontal diseases. The presence of 24p3R was examined in periodontal cells, including human gingival fibroblasts, periodontal ligament fibroblasts, human oral epithelial cells (HOECs), and neutrophil-like cells (HL-60), by Western blotting. Changes in periodontal disease-associated proteins in the presence of recombinant LCN2 (rLCN2) were examined using a protein array in differentiated HL-60 (dHL-60) cells. Interleukin-8 (IL-8) and macrophage inflammatory protein-1α (MIP-1α) mRNA expressions were analyzed by qRT-PCR, and IL-8 and MIP-1α levels in dHL-60 cells treated with rLCN2 or Porphyromonas gingivalis-lipopolysaccaharide (P.g-LPS) were determined by enzyme-linked immunosorbent assay. We detected 24p3R in dHL-60 cells. IL-8 was highly expressed and MIP-1α was weakly expressed in dHL-60 cells using a protein array. rLCN2 significantly decreased IL-8 mRNA and protein levels and suppressed P.g-LPS-induced IL-8 production in dHL-60 cells. As dHL-60 cells were co-cultured with HOECs in which LCN2 was knocked down, IL-8 mRNA expression increased in dHL-60 cells. Furthermore, rLCN2 inhibited MIP-1α production in dHL-60 cells. LCN2 suppresses inflammatory responses by regulating IL-8 and MIP-1α expression in periodontal diseases.

Role of Adenosine A1 Receptor in Sleep Deprivation-Induced Neuroinflammation: Insights on Rapid Eye Movement Sleep and Fear Extinction Memory Recall in Rats.

Sleep deprivation (SD), stemming from a myriad of aetiologies, is a prevalent health condition frequently overlooked. It typically impairs memory consolidation and synaptic plasticity, potentially through neuroinflammatory mechanisms and adenosinergic signalling. It is still unclear whether the adenosine A1 receptor (A1R) modulates SD-induced neurological deficits in the hippocampus. This study aims to evaluate the effects of SD on fear extinction memory recall and emotional behaviour in male Sprague Dawley rats; to investigate the role of A1R antagonism by the administration of 8-cyclopentyltheophylline (8-CPT), an A1R antagonist during 48-hour SD in mitigating neuroinflammation and synaptic plasticity deficits induced by SD; and to assess changes in hippocampal neurogenesis, neuronal cell death, and sleep architecture in response to A1R antagonism during SD. A total of 39 animals were used in the study, and they were divided into three experimental groups: 1) cage control (CC; n = 13); 2) SD for 48 hours (SD; n = 13); 3) SD for 48 hours+ 8-CPT (20 mg/kg/day in 20% DMSO divided into two doses, morning and evening, i.p.; n = 13). 'n' refers to the sample size/number of animals in each group. Rats were subjected to SD after cued fear extinction training for 48 hours followed by fear extinction memory recall test, anxious-depressive-like behaviours by open field test (OFT), sucrose preference test, and forced swim test (FST). Levels of adenosine in the hippocampus were quantified by high-performance liquid chromatography. Protein levels of interleukin-6 (IL-6) and IL-10 were quantified by enzyme-linked immunosorbent assay (ELISA). Expression levels of proteins and genes of interest were analysed using immunohistochemistry and real-time polymerase chain reaction (RT-PCR), respectively. Sleep architecture was assessed by recording electroencephalography (EEG), electromyography, and electrooculography from rats. Administration of CPT during SD reversed extinction recall impairments (p = 0.01), improved line crossings in OFT, sucrose preference (p < 0.01), and reduced immobility during the FST (p < 0.01). Immunohistochemical analysis of DG, CA3, and CA1 regions of the hippocampus revealed a significant upregulation of A1R expression in the SD and SD+CPT groups (p < 0.001, n = 5). Expression of post-synaptic density protein (PSD-95) and synaptophysin increased and a marked reduction in the Toll-like receptor-4(TLR-4) expression in activated microglia in the SD+CPT group. 8-CPT partially restored SD-induced decline in serotonin and brain-derived neurotrophic factor. SD-induced neuronal apoptosis through caspase-3 and the P-p38 mitogen-activated protein kinase pathway was partially reversed by 8-CPT. RT-PCR results showed that A1R antagonism attenuated gene expression of pro-inflammatory cytokines (IL-1β, TNFα, p-NFκB s536, and IL-6) and increased anti-inflammatory cytokines (IL-1ra, IL-4, IL-10, IL-11, and IL-13) during SD. EEG recordings revealed that A1R antagonism increased REM sleep without affecting non-REM sleep during SD, leaving rebound sleep unaffected. Conclusion: These findings highlight the role of A1R antagonism in restoring fear extinction memory recall, synaptic plasticity, adult neurogenesis, neuronal cell death, and attenuating neuroinflammation during SD, paving the way for the further exploration of its therapeutic potential in sleep-related cognitive deficits.

Comprehensive Analysis of circRNA and mRNA Revealing Potential Mechanism Underlying Neuroinflammation in BV2 Cells

Background: The significance of circular RNAs (circRNAs) in diabetic complications has been established. However, their role in basal and diabetic states, as well as cognitive dysfunction, requires further investigation. Methods: BV-2 microglial cells were exposed to high glucose (50mM) and insulin (2μM) for 48 hours. The levels of interleukin-1beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factoralpha (TNF-α) were assessed through quantitative polymerase chain reaction (qPCR), western blot, and ELISA. CircRNA and messenger RNA (mRNA) sequencing were performed, and the data were analyzed. Differentially expressed circRNAs and mRNAs were identified using qPCR. The circRNA-miRNA interaction was predicted using Miranda and TargetScan software, and their levels were quantified by qPCR. Results: The results demonstrated a significant increase in mRNA and protein levels of IL-1β, IL-6, and TNF-α in BV2 cells treated with glucose and insulin. Five circRNAs (four upregulated and one downregulated) were identified in both glucose and insulin groups compared to the control. Further qPCR analysis revealed marked increases in the levels of chr17:40159331- 40159711+ and chr2:72800499-72801858- (mmu_circ_0010164) in both treatment groups. Competitive endogenous RNA networks showed significant upregulation of mRNA levels of mitochondrial transcription termination factor 1b (Mterf1b) and G protein subunit gamma 4 (Gng4), accompanied by a decrease in mmu-miR-6918-3p and mmu-miR-7043-3p levels in the glucose and insulin groups compared to the control. Knockdown of mmu_circ_0010164 significantly inhibited the inflammatory response induced by glucose and insulin in BV-2 microglial cells. Conclusion: These findings indicate that both glucose and insulin can elicit inflammatory responses in BV2 cells through the modulation of mmu_circ_0010164 levels. The underlying mechanism may involve potential downstream targets of mmu_circ_0010164, specifically mmu-miR-7043-3p/Gng4 and mmu-miR-6918-3p/Mterf1b. This provides novel insights into the treatment of glucose-induced neuroinflammation.

Value of Serum Retinol-Binding Protein, Lipoprotein A and Inflammatory Nutritional Indexes and their Interactions in Predicting Acute Kidney Injury after Coronary Angiography

Background: Contrast-induced acute kidney injury (CI-AKI) is a serious condition that can arise after undergoing coronary angiography (CAG). This complication is primarily caused by the toxic impact of contrast agents on the kidneys, along with oxidative stress and inflammatory responses. Identifying reliable biomarkers for early prediction remains essential due to the limited accuracy of existing clinical risk scores. Methods: This retrospective case-control study included 365 patients undergoing CAG between December 2021 and December 2023. Patients were categorized into non-AKI (n = 320) and AKI (n = 45) groups based on post-procedural renal function. Biomarkers including serum retinol-binding protein (RBP), lipoprotein A (Lp(a)), interleukin-6 (IL-6), and nutritional markers were measured. Baseline demographics, lipid profiles, and procedure details were recorded. Results: Baseline characteristics revealed no significant differences between groups in terms of age, BMI, gender, and prevalence of hypertension or diabetes mellitus. Notably, RBP (41.83 ± 10.22 mg/L vs. 32.45 ± 8.76 mg/L, p &lt; 0.001), Lp(a) (42.35 ± 13.87 mg/dL vs. 35.76 ± 11.54 mg/dL, p &lt; 0.001), and IL-6 (6.23 ± 2.14 pg/mL vs. 4.87 ± 1.59 pg/mL, p &lt; 0.001) levels were significantly higher in the AKI group. Total protein levels were significantly lower in the AKI group (6.88 ± 0.62 g/dL vs. 7.12 ± 0.54 g/dL, p = 0.006). Multivariate logistic regression identified RBP (OR 1.118, 95% CI 1.072–1.166, p &lt; 0.001), Lp(a) (OR 1.047, 95% CI 1.014–1.080, p = 0.005), and IL-6 (OR 1.710, 95% CI 1.355–2.157, p &lt; 0.001) as independent predictors of AKI, while higher total protein levels were protective (OR 0.497, 95% CI 0.257–0.960, p = 0.037). Conclusion: Serum RBP, Lp(a), IL-6, and total protein levels are valuable biomarkers for predicting CI-AKI after CAG. Elevated RBP, Lp(a), and IL-6 indicate higher risk, while higher total protein suggests protective effects.

TMIC-36. EVALUATING PHARMACOLOGICAL INHIBITION OF LSD1 IN MODELS OF DIFFUSE MIDLINE GLIOMA AND EFFECTS ON IMMUNE MICROENVIRONMENT SIGNALING

Abstract BACKGROUND Diffuse midline glioma (DMG) is the leading cause of death for pediatric brain tumor patients. DMG is remarkably immunologically senescent, even in comparison to other “immune-cold” tumors like adult glioblastoma. Lysine specific demethylase-1 (LSD1) is a histone demethylase exerting context-specific mechanisms of gene repression and activation and can be inhibited pharmacologically. We found LSD1 regulates the expression of inflammatory cytokines such as Interleukin 18 (IL18) and immune signaling receptors in DMG. Here, we investigated whether IL18 is transcriptionally regulated by LSD1 in vitro and in vivo using clinically relevant small molecule inhibitors. METHODS LSD1 binding to the IL-18 promoter was evaluated using chromatin immunoprecipitation. Transcript and protein expression using qRT-PCR and western blotting assessed the impact of LSD1 inhibition on IL18-related pathways and immune signaling pathways. LSD1 inhibitors (IMG-7289 or GSKLSD1) were evaluated patient-derived xenograft models. Results. We found elevated LSD1 binding on the IL18 promoter in DIPG VI cells. Subsequent qRT-PCR and western blot analyses demonstrated increased IL18 protein levels following treatment with GSKLSD1 or IMG-7289 in DIPG VI cells, alongside heightened IL18 mRNA expression compared to DMSO-treated cells. Either GSKLSD1 or IMG-7289 treatment in DIPG IV cells also led to significantly elevated IL18 binding protein mRNA levels. Orthotopic xenografts using DIPG IV bearing mice dosed with LSD1 inhibitors over the course of 35 days several weeks showed a decrease in IL-18R, and an increase in IL-18 BP transcript levels alongside a significant increase in Slamf7 transcript levels. Some evidence of tumor inhibition was seen in male but not female mice treated with GSKLSD1. CONCLUSIONS Our investigation revealed IL18 as a direct target of LSD1 specifically in DIPG VI, and gender specific effects of GSKLSD1 treatment in vivo. Future studies will prioritize validating and elucidating the underlying mechanisms the therapeutic potential of LSD1 inhibition in DMG.

Association of IL-6 G-174C (rs1800795) variant with the susceptibility to hepatocellular carcinoma in patients with chronic hepatitis

AimAn ineffective immune response resulting from dysregulation of cytokine production might encourage viral persistence and cause chronic viral hepatitis to worsen. This study examined the relationship between alterations in interleukin-6 (IL-6) levels and the IL-6 − 174 G > C (rs1800795) polymorphism, as well as how this polymorphism affects the development and progression of chronic hepatitis brought on by hepatitis B (HBV) and hepatitis C (HCV) into hepatocellular carcinoma (HCC).Patients and methodsWhole blood samples from 126 Egyptian patients with HCC (111 with HCV and 15 with HBV), as well as 126 age- and sex-matched healthy individuals, were used to extract DNA. Using PCR-based allele-specific amplification (ASA), the existence of the IL-6 G-174C polymorphism was investigated. Additionally, each participant's serum IL-6 levels were determined using an enzyme-linked immunosorbent assay (ELISA).ResultsThe primary observations revealed that HCC patients had greater serum levels of IL-6 compared to the control groups (p < 0.001). Patients with the variant (CG and GG) genotype in the HCC group were found to have more disease severity indicated by higher levels of alpha-fetoprotein (AFP) and a higher ascites grade, as well as increased inflammatory activity as defined by higher levels of IL-6 and C-reactive protein (CRP) (p < 0.001 for both) in comparison to patients with the wild-type (CC) genotype (p < 0.001 and p = 0.002, respectively).ConclusionThe rs1800795 SNP in the IL-6 gene was associated with increased inflammatory activity and high levels of IL-6, indicating that this SNP may play a role in the development of HCC in Egyptian patients with chronic viral hepatitis.

Effect of Dendrobii Officinalis Caulis water extract on hyperviscosity rats based on TLR4/NF-κB/ICAM-1 signaling pathway

This study aims to reveal the effect and mechanism of Dendrobii Officinalis Caulis water extract on the rat model of hyperviscosity induced by a high-sugar, high-salt, and high-fat diet. Thirty-six male SD rats were randomized into normal, model, Compound Danshen Tablets(0.5 g·kg~(-1)), and low-, medium-, and high-dose(0.25, 0.5, and 1.0 g·kg~(-1), respectively) Dendrobii Officinalis Caulis water extract groups. Except the normal group, the remaining groups were fed with a high-salt and high-fat diet and received regular gavage of sucrose solution(20 g·kg~(-1)) for 12 consecutive weeks. The modeling was accompanied by gavage of corresponding drugs. During the experimental period, general signs(such as grip strength, automatic activity, vertigo time, and pain threshold), blood rheology, and blood flow in the tail microcirculation were measured. Flow cytometry was employed to detect the positive cells of intercellular cell adhesion molecule 1(ICAM-1/CD54) and vascular cell adhesion molecule 1(VCAM-1/CD106) in the peripheral blood and the Ca~(2+) fluorescence intensity. Enzyme-linked immunosorbent assay was employed to measure the levels of ICAM-1, VCAM-1, platelet endothelial cell adhesion molecule 1(PECAM-1/CD31), endothelin 1(ET-1), thromboxane A2(TXA2), prostaglandin I2(PGI2), nitric oxide(NO), interleukin-1β(IL-1β), C-reactive protein(CRP), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α), and lipopolysaccharide(LPS). Hematoxylin-eosin, Masson, and Gomori's aldehyde-fuchsin staining were used to observe the pathological and morphological changes in rat colon and aorta. Fluorescence quantitative PCR was conducted to determine the mRNA levels of Toll-like receptor 4(TLR4), myeloid differentiation factor 88(MyD88), nuclear factor-kappaB(NF-κB), ICAM-1, and VCAM-1 in the aortic tissue. Western blot was employed to determine the protein levels of occludin and claudin-1 in the colon tissue and ICAM-1, VCAM-1, and IL-6 in the aortic tissue. Immunohistochemistry and immunofluorescence were employed to detect the expression of TLR4 and NF-κB in the aortic tissue. The results showed that Dendrobii Officinalis Caulis water extract significantly improved the general signs, reduced the whole blood viscosity, whole blood reducing viscosity, and whole blood flow resistance, increased the blood flow of tail microcirculation, reduced the rates of CD54, CD106-positive cells in peripheral blood, and attenuated the blood flow obstruction in the rat model of hyperviscosity. The water extract significantly lowered the serum levels of ICAM-1, VCAM-1, PECAM-1, and ET-1, elevated the PGI2 and NO level, and alleviated vascular endothelial injury. In addition, the water extract decreased the content of IL-1β, TNF-α, IL-6, and CRP in the serum and reduced the vascular inflammatory injury. Moreover, the Dendrobii Officinalis Caulis water extract improved the morphology of colon and aortic tissues, promoted the expression of occludin and claudin-1 in the colon, and down-regulated the mRNA levels of TLR4, MyD88, NF-κB, ICAM-1, and VCAM-1 as well as the protein levels of TLR4, NF-κB, and IL-6 in the aortic tissue. In conclusion, the results suggest that the Dendrobii Officinalis Caulis water extract may regulate the endothelial inflammatory injury via the TLR4/NF-κB/ICAM-1 pathway, thereby ameliorating hyperviscosity.

Alpha(α)-mangostin (Xanthone of Garcinia mangostana L.): Augmenting Macrophages Activity for an Effective Diabetic Wound Healing

Diabetic wounds are particularly difficult to treat medically because they heal at a slower pace than regular wounds. Macrophages are essential in all stages of normal wound healing, including inflammation, proliferation and rem odelling. When wound healing is affected, macrophages can reduce the level of growth factor and increase the level of interleukin-6 (IL-6), disrupt the balance between tissue inhibitors of metalloproteinase (TIMPs) and matrix metalloproteinase (MMPs), which can slow down healing. Alpha(α)-mangostin, a natural xanthone derived from the pericarp of the mangosteen, has gained considerable attention due to its anti-inflammatory properties, suggesting its potential to promote wound healing. However, its exact role in healing diabetic foot ulcers, common in diabetes, remains unclear. Hence, this study aims to explore how α-mangostin might affect diabetic wound healing by evaluating its impact on PDGF, CTGF, BFGF, VEGF, TGF-β, MMP-9, TIMP-2 and IL-6 secretion in macrophage cells. Human monocytic macrophages (THP-1) were incubated with a 35 mM glucose solution for 72 h to create a glucose-enriched medium. The cells were then incubated with α-mangostin (0.15, 2.5 and 5 µg/mL) together with 35 mM glucose. Carboxymethyl cellulose (CMC) served as positive controls; glucose-enriched media and media-alone served as negative controls. Protein expression was measured using ELISA. α-mangostin (2.5 µg/mL) increased the levels of PDGF and VEGF and decreased the level of MMP-9 compared to glucose controls. There was no significant difference in other growth factors, TIMP-2 and IL-6 protein levels across any of the treatment groups compared to glucose controls. In conclusion, α-mangostin particularly at 2.5 µg/mL demonstrated a significant increase in PDGF and VEGF levels while simultaneously reducing MMP-9 in macrophage cells under glucose-induced conditions. These findings suggest that α-mangostin holds the potential for enhancing the healing of chronic wounds in diabetic conditions. HIGHLIGHTS Treatment of macrophages (THP-1 cells) with 2.5 µg/mL Alpha (α)-mangostin in a high glucose medium led to a significant increase in PDGF and VEGF secretion compared to glucose control. Treatment of the cells with 2.5 µg/mL Alpha (α)-mangostin in a high glucose medium led to a significant reduction of MMP-9 secretion compared to glucose control. There is no significant effects of Alpha (α)-mangostin on IL-6 and TIMP secretion compared to controls This study suggests that Alpha (α)-mangostin may be beneficial in promoting diabetic wound healing via stimulation of PDGF and VEGF secretion and reducing MMP-9 levels. GRAPHICAL ABSTRACT

Chromosomal instability is associated with prognosis and efficacy of bevacizumab after resection of colorectal cancer liver metastasis

Introduction Individualized treatment of colorectal cancer liver metastases (CRLM) remains challenging due to differences in the severity of metastatic disease and tumour biology. Exploring specific prognostic risk subgroups is urgently needed. The current study aimed to investigate the prognostic value of chromosomal instability (CIN) in patients with initially resectable CRLM and the predictive value of CIN for the efficacy of bevacizumab. Methods Ninety-one consecutive patients with initially resectable CRLM who underwent curative liver resection from 2006 to 2018 at Sun Yat-sen University Cancer Center were selected for analysis. CIN was evaluated by automated digital imaging systems. Immunohistochemistry (IHC) was performed to detect interleukin-6 (IL-6), vascular endothelial growth factor A (VEGFA) and CD31 expression in paraffin-embedded specimens. Recurrence-free survival (RFS) and overall survival (OS) were analysed using the Kaplan–Meier method and Cox regression models. Results Patients with high chromosomal instability (CIN-H) had a worse 3-year RFS rate (HR, 1.953; 95% CI, 1.001–3.810; p = 0.049) and a worse 3-year OS rate (HR, 2.449; 95% CI, 1.150–5.213; p = 0.016) than those with low chromosomal instability (CIN-L). CIN-H was identified as an independent prognostic factor for RFS (HR, 2.569; 95% CI, 1.078–6.121; p = 0.033) and OS (HR, 3.852; 95% CI, 1.173–12.645; p = 0.026) in the multivariate analysis. The protein levels of IL-6, VEGFA and CD31 were upregulated in patients in the CIN-H group compared to those in the CIN-L group in both primary tumour and liver metastases tissues. Among them, 22 patients with recurrent tumours were treated with first-line bevacizumab treatment and based on the clinical response assessment, disease control rates were adversely associated with chromosomal instability (p = 0.043). Conclusions Our study showed that high chromosomal instability is a negative prognostic factor for patients with initially resectable CRLM after liver resection. CIN may have positive correlations with angiogenesis through expression of IL-6–VEGFA axis and be used as a potential predictor of efficacy of bevacizumab.

Study of Association of Infection-related Biomarkers and Inflammatory Cytokines with the Severity of Coronavirus Disease 2019: A Prospective Observational Study.

Sepsis-linked biomarkers and inflammatory cytokines are markedly associated with potential risks of progression to severity in coronavirus disease 2019 (COVID-19). Clinical studies that find a plausible association between sepsis biomarkers and the inflammatory cytokine response in the Indian community need to be studied with clarity. To study the relationship between sepsis-linked biomarkers and inflammatory cytokines interleukin-6 (IL-6), C-reactive protein (CRP), ferritin, and D-dimer linked to clinical severity resulting from COVID-19 infection. The present prospective observational cohort study was conducted between March and December 2021 in the Department of Critical Care Medicine at a tertiary care hospital in Pune, Maharashtra, India, on COVID-19 patients. Upon patient admission, inflammatory biomarkers such as IL-6, CRP, ferritin, and D-dimer were recorded. Oxygen requirements during hospitalization, invasive mechanical ventilation (IMV), high-flow nasal cannula (HFNC), noninvasive mechanical ventilation (NIV), duration of ventilator use, intensive care unit (ICU) stay, and mortality were documented. The average levels of IL-6, CRP, D-dimer, and serum ferritin protein recorded at the time of patient arrival were notably higher in the severe (S) group compared to the nonsevere (NS) group. The average duration of ventilator use, ICU stay, and hospital stay was significantly longer in the S group than in the NS group. The percentage of patients who required HFNC, NIV, IMV, and mortality was significantly higher in the S group compared to the NS group. Sepsis-linked biomarkers and inflammatory cytokines such as IL-6, CRP, D-dimer, and serum ferritin levels at the time of admission were markedly associated with severity outcomes in COVID-19 infection.

The role of interleukin-10 in mitigating endoplasmic reticulum stress in aged mice through exercise.

Although unfolded protein response (UPR) is essential for cellular protection, its prolonged activation may induce apoptosis, compromising cellular longevity. The aging process increases the endoplasmic reticulum (ER) stress in skeletal muscle. However, whether combined exercise can prevent age-induced ER stress in skeletal muscle remains unknown. Evidence suggests that ER stress may increase inflammation by counteracting the positive effects of interleukin-10 (IL-10), whereas its administration in cells inhibits ER stress and apoptosis. This study verified the effects of aging and combined exercise on physical performance, ER stress markers, and inflammation in the quadriceps of mice. Moreover, we verified the effects of IL-10 on ER stress markers. C57BL/6 mice were distributed into young (Y, 6 mo old), old sedentary (OS, sedentary, 24 mo old), and old trained group (OT, submitted to short-term combined exercise, 24 mo old). To clarify the role of IL-10 in UPR pathways, knockout mice lacking IL-10 were used. The OS mice presented worse physical performance and higher ER stress-related proteins, such as C/EBP homologous protein (CHOP) and phospho-eukaryotic translation initiation factor 2 alpha (p-eIF2α/eIF2α). The exercise protocol increased muscle strength and IL-10 protein levels in OT while inducing the downregulation of CHOP protein levels compared with OS. Furthermore, mice lacking IL-10 increased BiP, CHOP, and p-eIF2α/eIF2α protein levels, indicating this cytokine can regulate the ER stress response in skeletal muscle. Bioinformatics analysis showed that endurance and resistance training downregulated DNA damage inducible transcript 3 (DDIT3) and XBP1 gene expression in the vastus lateralis of older people, reinforcing our findings. Thus, combined exercise is a potential therapeutic intervention for promoting adjustments in ER stress markers in aged skeletal muscle.NEW & NOTEWORTHY Aging elevates endoplasmic reticulum (ER) stress in skeletal muscle, potentially heightening inflammation by opposing interleukin-10 (IL-10) effects. This study found that short-term combined exercise boosted strength and IL-10 protein levels while reducing CHOP protein levels in older mice. In addition, IL-10-deficient mice exhibited increased ER stress markers, highlighting IL-10's role in regulating ER stress in skeletal muscle. Consequently, combined exercise emerges as a therapeutic intervention to elevate IL-10 and adjust ER stress markers in aging.

Effects of Luhong Yixin Granules on myocardial fibrosis in heart failure rats based on TGF-β1/Smads signaling pathway

To investigate the effects of Luhong Yixin Granules on myocardial fibrosis in rats with heart failure and its possible mechanism, a total of 60 male Wistar rats were randomly divided into the control group, model group, and low-, medium-and high-dose Luhong Yixin Granules groups, with 12 rats in each group. Except for those in the control group, rats in the other groups were induced by intraperitoneal injection of doxorubicin(DOX) into a rat model. After the Luhong Yixin Granules were dissolved in the same amount of normal saline, they were given by gavage at low, medium and high doses(2.8, 5.6, 11.2 g·kg~(-1)·d~(-1)), and the control group and the model group were given the same amount of normal saline by gavage for 40 days. After the end of dosing, echocardiography was used to measure left ventricular ejection fraction(LVEF) and left ventricular fractional shortening(LVFS). Rat body weight(BW) and heart weight(HW) were calculated as HW/BW. Enzyme-linked immunosorbent assay was used to measure the levels of interleukin-6(IL-6), interleukin-17(IL-17), tumor necrosis factor-α(TNF-α), transforming growth factor-β1(TGF-β1), growth stimulation expressed gene 2 protein(ST2), N-terminal pro-B-type natriuretic peptide(NT-proBNP), galectin-3(Gal-3) and creatine kinase isoenzyme(CK-MB) in serum. Hematoxylin-eosin(HE) staining and Masson staining were used to observe the pathological morphology of myocardial tissue. Western blot and quantitative real-time polymerase chain reaction were used to detect the protein and mRNA expression levels of IL-6, IL-17, TNF-α, TGF-β1, Smad3, Smad7, α-smooth muscle actin(α-SMA), and collagen Ⅰ(COL-Ⅰ), respectively. RESULTS:: showed that compared with those in the control group, LVEF, LVFS, and HW/BW in the model group were decreased(P&lt;0.05), and the levels of IL-6, IL-17, TNF-α, TGF-β1, ST2, NT-proBNP, Gal-3, and CK-MB were increased(P&lt;0.05). HE staining showed inflammatory changes in myocardial tissue; Masson staining showed decreases in the cross-sectional area and ventricular cavity area of the heart, and myocardial fibrosis of varying degrees(P&lt;0.05). The protein and mRNA expression of IL-6, IL-17, TNF-α, TGF-β1, Smad3, α-SMA, and COL-Ⅰ were increased(P&lt;0.05), and the protein and mRNA expression of Smad7 protein was decreased(P&lt;0.01). Compared with those in the model group, LVEF, LVFS and HW/BW of the low-, medium-and high-dose Luhong Yixin Granules groups were increased(P&lt;0.05), and the levels of IL-6, IL-17, TNF-α, TGF-β1, ST2, NT-proBNP, Gal-3 and CK-MB were decreased(P&lt;0.05). HE staining showed gradually reduced inflammatory changes of myocardial tissue, and Masson staining showed increased cross-sectional area and ventricular cavity area of the heart and decreased area of myocardial fibrosis(P&lt;0.05). The protein and mRNA expression levels of IL-6, IL-17, TNF-α, TGF-β1, Smad3, α-SMA, and COL-Ⅰ were decreased(P&lt;0.05), while the protein and mRNA expression levels of Smad7 were increased(P&lt;0.05). Luhong Yixin Granules may be of great value in the treatment of heart failure by regulating the TGF-β1/Smads signaling pathway, inhibiting the expression of inflammation-related proteins, reducing the deposition of extracellular matrix, and alleviating myocardial fibrosis.

Dual analysis of IL-8 in non-small cell lung cancer: A clinical perspective on protein and RNA evaluation via imaging mass cytometry.

e15068 Background: The cytokine Interleukin-8 (IL-8) plays a pivotal role in the tumorigenesis and progression of non-small cell lung cancer (NSCLC), influencing both the tumor microenvironment and immune escape mechanisms. Given its significance, accurately assessing IL-8 levels at the protein and RNA levels could provide valuable insights into its prognostic value and inform therapeutic targeting strategies. This study utilizes imaging mass cytometry (IMC) to conduct a comprehensive evaluation of IL-8 expression in NSCLC, integrating both protein presence and gene expression analysis within the spatial context of the tumor microenvironment. Methods: Our approach leverages the capabilities of IMC to simultaneously detect protein and RNA levels of IL-8 in NSCLC tissue samples. For protein analysis, antibodies tagged with heavy metal isotopes specifically target IL-8, allowing for precise quantification and spatial localization within the tumor. Concurrently, RNA detection is achieved through hybridization probes designed for IL-8 mRNA, enabling the direct observation of gene expression patterns. This method provides a unique opportunity to correlate IL-8's protein abundance with its mRNA expression, offering insights into the regulatory mechanisms at play and their impact on the tumor's behavior and patient outcomes. Results: Preliminary data demonstrate a marked heterogeneity in IL-8 distribution at both the protein and RNA levels, with distinct patterns emerging that relate to tumor aggressiveness and immune infiltration. High IL-8 expression, identified through our integrated IMC approach, correlates with poor prognosis and reduced response to standard therapies, underscoring its potential as a biomarker for NSCLC management. Furthermore, the spatial analysis afforded by IMC highlights the microenvironmental niches where IL-8-mediated interactions could drive tumor progression and immune evasion. Conclusions: By employing IMC for the dual analysis of IL-8 protein and RNA in NSCLC, our study provides a comprehensive view of this cytokine's role in lung cancer pathology. The ability to visualize and quantify IL-8 expression within the spatial context of the tumor microenvironment opens new avenues for understanding its biological impact and for developing targeted therapies. Our findings advocate for the inclusion of IL-8 in the biomarker panel for NSCLC, with potential implications for patient stratification and personalized treatment planning. Future research will focus on expanding these observations to a larger cohort and exploring the therapeutic modulation of IL-8 as a strategy for improving NSCLC outcomes.

LncRNA NEAT1/miR-211/IL-10 Axis Regulates Inflammation of Peripheral Blood Mononuclear Cells in Acute Myocardial Infarction

This study aimed to explore the expression of long non-coding RNA (lncRNA) nuclear paraspeckle assembly transcript 1 (NEAT1) in patients with acute myocardial infarction (AMI) and its inflammatory regulation mechanism through miR-211/interleukin 10 (IL-10) axis.A total of 75 participants were enrolled in this study: 25 healthy people in the control group, 25 patients with stable angina pectoris (SAP) in the SAP group, and 25 patients with AMI in the AMI group. Real-time qPCR was used to detect mRNA expression levels of NEAT1, miR-211, and IL-10. The interaction between miR-211, NEAT1, and IL-10 was confirmed by dual-luciferase reporter assay, and protein expression was detected using western blot.High expression of NEAT1 in peripheral blood mononuclear cells (PBMCs) of patients with AMI was negatively related to serum creatine kinase-MB (CK-MB), cardiac troponin I (cTnI), tumor necrosis factor-α (TNF-α), IL-6, and IL-1β and was positively correlated with left ventricular ejection fraction (LVEF). In THP-1 cells, miR-211 was confirmed to target and inhibit IL-10 expression. NEAT1 knockdown and miR-211-mimic markedly decreased IL-10 protein levels, whereas anti-miR-211 markedly increased IL-10 protein levels. Importantly, miR-211 level was negatively related to NEAT1 and IL-10 levels, whereas IL-10 level was positively related to the level of NEAT1 expression in PBMCs of patients with AMI.LncRNA NEAT1 was highly expressed in PBMCs of patients with AMI, and NEAT1 suppressed inflammation via miR-211/IL-10 axis in PBMCs of patients with AMI.

Retinoic Acid Neutralizes the Effects of Herpes Simplex Virus Type 1-Infected Cell Protein 0 (ICP0) in Retinal Pigment Epithelial Cells.

Herpes simplex virus (HSV) infection of the cornea, uvea, and retina is the leading infectious cause of blindness worldwide. This study examined the effects of retinoic acid (RA) on the protein levels of interleukin (IL)-17A and IL-23 cytokines with known proinflammatory effects and toll-like receptor 3 (TLR3) messenger RNA (mRNA) expression in retinal pigment epithelial (ARPE-19) cells treated with HSV-1-infected cell protein 0 (ICP0). We used 3-[4.5-dimethylthiazol-2-yl]-2.5-diphenyl tetrazolium bromide assay to calculate the half maximal inhibitory concentration (IC50) doses of RA and ICP0 in ARPE-19 cells. At the end of 24 hours, protein levels of IL-17A and IL-23 were analyzed using enzyme-linked immunosorbent assay. TLR3 mRNA expression levels were also calculated using reverse transcription-polymerase chain reaction (RT-PCR). RA administration decreased IL-17A levels, which were elevated by ICP0. The IL-23 levels were similar between the ICP0-treated and control groups, but the difference was significant between the ICP0-treated group and RA+ICP0 combination. These results showed that RA can significantly increase IL-23 levels in the presence of ICP0. Although ICP0 dramatically increased TLR3 mRNA expression compared with that in the control group, the RA+ICP0 combination returned TLR3 mRNA expression to a level similar to that in the control group (P = 0.419). RA may potentially neutralize HSV-1 ICP0 negative effects in ARPE-19 cells.

Decreased Hepatic and Serum Levels of IL-10 Concur with Increased Lobular Inflammation in Morbidly Obese Patients.

Background and Objectives: Non-alcoholic fatty liver disease (NAFLD) is associated with obesity and ranges from simple steatosis to non-alcoholic steatohepatitis (NASH), fibrosis, cirrhosis, and hepatocellular carcinoma. Accumulating evidence in animal models suggests that loss of interleukin-10 (IL-10) anti-inflammatory actions might contribute to lobular inflammation, considered one of the first steps toward NASH development. However, the role of IL-10 in lobular inflammation remains poorly explored in humans. We examined mRNA and protein levels of IL-10 in liver biopsies and serum samples from morbidly obese patients, investigating the relationship between IL-10 and lobular inflammation degree. Materials and Methods: We prospectively enrolled morbidly obese patients of both sexes, assessing the lobular inflammation grade by the Brunt scoring system to categorize participants into mild (n = 7), moderate (n = 19), or severe (n = 13) lobular inflammation groups. We quantified the hepatic mRNA expression of IL-10 by quantitative polymerase chain reaction and protein IL-10 levels in liver and serum samples by Luminex Assay. We estimated statistical differences by one-way analysis of variance (ANOVA) and Tukey's multiple comparison test. Results: The hepatic expression of IL-10 significantly diminished in patients with severe lobular inflammation compared with the moderate lobular inflammation group (p = 0.01). The hepatic IL-10 protein levels decreased in patients with moderate or severe lobular inflammation compared with the mild lobular inflammation group (p = 0.008 and p = 0.0008, respectively). In circulation, IL-10 also significantly decreased in subjects with moderate or severe lobular inflammation compared with the mild lobular inflammation group (p = 0.005 and p < 0.0001, respectively). Conclusions: In liver biopsies and serum samples of morbidly obese patients, the protein levels of IL-10 progressively decrease as lobular inflammation increases, supporting the hypothesis that lobular inflammation develops because of the loss of the IL-10-mediated anti-inflammatory counterbalance.

Amygdalin ameliorates alopecia areata on C3H/HeJ mice by inhibiting inflammation through JAK2/STAT3 pathway

BackgroundEvidence has demonstrated that Chinese medicine formula Xuefu Zhuyu decoction can markedly promote the formation of new hair in patients and mice with alopecia areata (AA). Amygdalin is one of the active components of Xuefu Zhuyu decoction, but its therapeutic effects and the underlying mechanisms on AA remains largely unrevealed. PurposeTherefore, this study aims to investigate the therapeutic effects and to probe its molecular mechanisms of inflammation and immune regulation on AA model of C3H/HeJ mice. Study designThe C3H/HeJ female mice were divided into control, AA, rusolitinib (60 mg/kg), and amygdalin groups (60, 90, and 120 mg/kg, 0.2 ml/10 g, i.g.). MethodsThe optical microscope was used to observe the feature of the local skin, and the number of lanugo and terminal hair. H&E staining was performed to determine the degree of pathological damage to the skin. ELISA was performed to detect levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ) in mice serum. Flow cytometry was carried out to analyze the CD4+CD25+FOXP3+, CD4+ and CD8+ of skin tissue. And the levels of CD4+ and CD8+, p-JAK/JAK2, p-STAT3/STAT, and SOCS3 were detected by immunohistochemistry. Western blot and qRT-PCR were employed to examine the expression levels of IL-6, TNF-α, IFN-γ, JAK2, p-JAK, STAT, p-STAT3 and SOCS3 proteins and genes in skin tissues. ResultsCompared with AA group, amygdalin immensely increased the number of vellus hairs and decreased the number of terminal hairs determined by skin microscopy and H&E staining. ELISA, Western blot and qRT-PCR data showed that the levels of IL-6, TNF-α and IFN-γ in serum and skin tissues of AA mice were significantly increased, while amygdalin administration dramatically restrained the contents of the three pro-inflammatory factors. Flow cytometry and immunohistochemistry hinted that amygdalin observably enhanced the number of CD4+CD25+FOXP3+ and CD4+ cells, while inhibited the number of CD8+ positive cells in mice with AA. Moreover, amygdalin signally reduced JAK2/STAT3 pathway-related protein and gene levels in AA mice. ConclusionAmygdalin could inhibit inflammatory response and improve immune function in the treatment of AA. The underlying molecular mechanism may be related to inhibition of JAK2/STAT3 pathway.

Describing the Cellular Impact of IQOS™ Smoke Extract and Vibration on Human Vocal Fold Fibroblasts

ObjectivesThe isolated or combined effects of vibration and smoke extract (SE) from the IQOS™ “heat-not-burn” technology on human vocal fold fibroblasts (hVFF) were evaluated in an in vitro setting in order to elucidate their influence on vocal fold (patho-) physiology. Study DesignExperimental pilot study using intervention with IQOS™-SE in vitro. MethodsImmortalized hVFF were exposed to IQOS™-SE or control medium under static or vibrational conditions. A phonomimetic bioreactor was used to deliver vibrational patterns to hVFF over a period of 5 days. Cytotoxicity was quantified by lactate dehydrogenase assay. Effects on extracellular matrix production, inflammation, fibrogenesis, and angiogenesis were assessed by reverse transcription-quantitative polymerase chain reaction, Western Blot, enzyme-linked immunosorbent assay, and Magnetic Luminex assays. ResultsWe observed significant changes induced either by IQOS™-SE exposure alone (matrix metalloproteinase 1, fibronectin, cyclooxygenase (COX)1, interleukin-8 gene expression), or by the combination of IQOS™-SE and vibration (hyaluronidase 2, COX2, interleukin-8 protein levels, vascular endothelial growth factor D). ConclusionShort-term in vitro exposure of hVFF to IQOS™-SE did not result in cytotoxicity and reduced the gene expression of measured inflammation mediators, but had no effect on their protein expression. However, the clinical effects of long-term IQOS™ use are still not known and further research is needed in order to asses, if IQOS™ is in fact less harmful than conventional cigarettes.

Enoxaparin Effect on Interleukin-10 Levels in Iraqi Patients with COVID-19: A Case-Control Study.

Coronavirus disease 19 (COVID-19), an infectious disease resulting from a virus known as severe acute respiratory syndrome coronavirus 2 (SARS-COV-2), was discovered in China in 2019 and causes several mild to moderate respiratory conditions. This study aimed to reveal the changes in serum interleukin-10 (IL-10) and other parameters in Iraqi COVID-19 patients compared with healthy controls by studying the effects of enoxaparin and evaluating the potential of IL-10 as a disease activity marker. This was a case-control study that included 180 samples: 90 patients hospitalized with COVID-19 from November 2022 to 20 April 2023 (40 patients had never used enoxaparin, whereas 50 patients had taken enoxaparin) and 90 healthy, age- and sex-matched control. There were 44 female patients and 46 male patients. The mean age of the patients and controls was 53.8 years vs. 50.8 years, respectively. The sandwich enzyme-linked immunosorbent assay (ELISA) method was used to measure IL-10 levels, while other parameters were assessed using the colorimetric method. The results of the study indicated highly significant changes between the patients and healthy controls in IL-10, D-dimer, and C-reactive protein (CRP) levels, as well as liver and renal functions. These findings elucidated a significant change between enoxaparin patients and non-enoxaparin patients in IL-10, D-dimer, and CRP levels. However, the liver and renal functions were not significantly altered. The Spearman's rank correlation test investigated the relationship between serum IL-10 and CRP. The results displayed a strong positive relationship between IL-10 and CRP. There were no significant differences between the other analyzed parameters; consequently, the patients had higher concentrations of IL-10, D-dimer, and some other parameters than the healthy controls. Additionally, IL-10 may be used as a marker of disease activity. Enoxaparin will likely help control IL-10 and D-dimer concentrations in patients since IL-10 levels decreased in patients treated with enoxaparin.

Time-Dependent Sequential Changes of IL-10 and TGF-β1 in Mice with Deep Vein Thrombosis.

To detect the expression changes of interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) during the development of deep vein thrombosis in mice, and to explore the application value of them in thrombus age estimation. The mice in the experimental group were subjected to ligation of inferior vena cava. The mice were sacrificed by excessive anesthesia at 1 d, 3 d, 5 d, 7 d, 10 d, 14 d and 21 d after ligation, respectively. The inferior vena cava segment with thrombosis was extracted below the ligation point. The mice in the control group were not ligated, and the inferior vena cava segment at the same position as the experimental group was extracted. The expression changes of IL-10 and TGF-β1 were detected by immunohistochemistry (IHC), Western blotting and real-time qPCR. IHC results revealed that IL-10 was mainly expressed in monocytes in thrombosis and TGF-β1 was mainly expressed in monocytes and fibroblast-like cells in thrombosis. Western blotting and real-time qPCR showed that the relative expression levels of IL-10 and TGF-β1 in each experimental group were higher than those in the control group. The mRNA and protein levels of IL-10 reached the peak at 7 d and 10 d after ligation, respectively. The mRNA expression level at 7 d after ligation was 4.72±0.15 times that of the control group, and the protein expression level at 10 d after ligation was 7.15±0.28 times that of the control group. The mRNA and protein levels of TGF-β1 reached the peak at 10 d and 14 d after ligation, respectively. The mRNA expression level at 10 d after ligation was 2.58±0.14 times that of the control group, and the protein expression level at 14 d after ligation was 4.34±0.19 times that of the control group. The expressions of IL-10 and TGF-β1 during the evolution of deep vein thrombosis present time-dependent sequential changes, and the expression levels of IL-10 and TGF-β1 can provide a reference basis for thrombus age estimation.