Interleukin-1 Activity Research Articles

An NLR family member X1 mutation (p.Arg707Cys) suppresses hepatitis B virus infection in hepatocytes and favors the interaction of retinoic acid-inducible gene 1 with mitochondrial antiviral signaling protein.

NLR family member X1 (NLRX1) is an important member of the NOD-like receptor (NLR) family and plays unique roles in immune system regulation. Patients with hepatitis B virus (HBV) infection are more likely to have the NLRX1 mutation p.Arg707Cys than healthy individuals. It has been reported that NLRX1 increases the infection rate of HBV in HepG2 cells expressing sodium taurocholate cotransporting polypeptide (NTCP). However, the role of NLRX1 mutation (p.Arg707Cys) in hepatitis remains unclear. We constructed Huh7 cells that stably overexpressed NTCP, using LV003 lentivirus. First, wild-type (WT) and mutant (MT) NLRX1 overexpression plasmids were constructed. The MT plasmid contained a point mutation at position 707 of the WT overexpression plasmid. Then, Huh7-NTCP cells were transfected with the WT or MT NLRX1 overexpression plasmid, and subsequent NLRX1 expression was analyzed using real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. HBV RNA levels were determined using RT-qPCR. HBsAg and HBcAg levels were confirmed immunohistochemically. Interferon alpha (IFN-α), interleukin 6 (IL-6), and type I interferon beta (IFN-β) levels were determined using enzyme-linked immunosorbent assay kits. p-p65, p-interferon regulatory factor (IRF) 3, and p-IRF7 expression levels were examined using western blot. The interaction of NLRX1 and retinoic acid-inducible gene (RIG)-1/mitochondrial antiviral signaling (MAVS) protein was confirmed by coimmunoprecipitation. The interaction of NLRX1 with IFN-α, IL-6, or IFN-β was analyzed by dual luciferase reporter gene assay. The levels of HBV RNA, HBsAg, and HBcAg in infected cells transfected with the WT NLRX1 or MT NLRX1 expression plasmid were higher than those in the untransfected control group; and these levels were lower in the cells transfected with MT NLRX1 than in those transfected with WT NLRX1. The levels of IFN-α, IFN-β, IL-6, p-p65, p-IRF3, and p-IRF7 were lower in cells transfected with WT NLRX1 or MT NLRX1 than in control cells. The levels of IFN-β, p-p65, p-IRF3, and p-IRF7 were higher in cells transfected with MT NLRX1 than in those transfected with WT NLRX1. Moreover, NLRX1 competitively inhibited RIG1 binding to MAVS, but the mutation in MT NLRX1 reduced this inhibitory effect. In addition, NLRX1 decreased the promoter activity of IFN-α, IFN-β, and IL-6. Our findings revealed that NLRX1 is a regulatory factor that inhibits the anti-HBV ability of hepatocytes and that the mutation p.Arg707Cys in NLRX1 suppresses HBV infection and activates the IFN/nuclear factor κB pathway.

Effects of anakinra on patients with heart failure below and above 60 years of age

Abstract Background Anakinra, an interleukin-1 (IL-1) blocker, is being explored in heart failure (HF) to improve cardiorespiratory fitness (CRF). CRF declines with increasing age. Purpose We aimed to determine the influence of anakinra on CRF in aging patients with HF. Methods We analyzed data from 73 patients with HF (55 [51-62] years, 37 (51%) female) treated with anakinra 100 mg subcutaneously daily. Patients were stratified into two groups according to a predetermined cut-off age of 60 years. We measured peak oxygen consumption (peak VO2) and C-reactive protein (CRP), a surrogate for IL-1 activity. Results We included 49 (67%) patients with age <60 years and 24 (33%) with age ≥60 years. The duration of treatment with anakinra was 4 [2-12] weeks. Age was negatively correlated with peak VO2 at baseline and during treatment with anakinra (Spearman’s Rho= -0.375, p=0.001 and Rho= -0.334, p=0.004, respectively). Younger patients had higher baseline peak VO2 (15.2 [12.4–17.7] vs 12.4 [10.3–14.3] mLO2·kg-1·min-1, p=0.001) but similar CRP (6.6 [3.6-16.6] vs 5.1 [2.7-11.2] mg/L, p=0.18). In both groups, anakinra decreased CRP (<60 years: from 6.6 [3.6–16.6] to 1.5 [0.9–3.8] mg/L, p<0.001; ≥60 years: from 5.1 [2.7–11.2] to 1.0 [0.5–3.4] mg/L, p<0.001)(Figure 1A) and increased peak VO2 (<60 years: from 15.2 [12.4–17.7] to 16.0 [14.0–18.7] mLO2·kg-1·min-1, p=0.036; ≥60 years: from 12.4 [10.3–14.3] to 13.7 [11.7–15.8] mLO2·kg-1·min-1, p<0.001)(Figure 1B). No significant differences in interval changes were observed between the groups. Conclusions Older patients with HF have a greater impairment in CRF than younger patients, yet they appear to benefit similarly from treatment with anakinra.Figure 1

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.

Molecular Mechanisms of Skatole-Induced Inflammatory Responses in Intestinal Epithelial Caco-2 Cells: Implications for Colorectal Cancer and Inflammatory Bowel Disease.

Inflammatory cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6), in intestinal epithelial cells significantly contribute to inflammatory bowel disease (IBD) and colorectal cancer (CRC). Given our previous findings that TNF-α is upregulated in intestinal epithelial Caco-2 cells induced by skatole, a tryptophan-derived gut microbiota metabolite, the present study aimed to explore the relationship between skatole and IL-6, alongside TNF-α. Skatole elevated the promoter activity of IL-6 as well as TNF-α, and increased IL-6 mRNA expression and protein secretion. In addition to activating NF-κB, the NF-κB inhibitor BAY 11-7082 reduced skatole-induced cell survival and the mRNA expression of IL-6 and TNF-α. NF-κB activation was attenuated by the extracellular signal-regulated kinase (ERK) pathway inhibitor U0126 and the p38 inhibitor SB203580, but not by the c-Jun N-terminal kinase (JNK) inhibitor SP600125. U126 and SB203580 also decreased the skatole-induced increase in IL-6 expression. When skatole-induced AhR activation was inhibited by CH223191, in addition to promoting NF-κB activation, IL-6 expression was enhanced in a manner similar to that previously reported for TNF-α. Taken together, these results suggest that skatole-elicited NF-κB activation induces IL-6 and TNF-α expression, although AhR activation partially suppresses this process. The ability of skatole to increase the expression of IL-6 and TNF-α may significantly affect the development and progression of these diseases. Moreover, the balance between NF-κB and AhR activation appears to govern the skatole-induced increases in IL-6 and TNF-α expression. Therefore, the present findings provide new insights into the mechanisms linking tryptophan-derived gut microbiota metabolites with colorectal disease.

The Antidiabetic Drug Metformin Attenuated Depressive and Anxietylike Behaviors and Oxidative Stress in the Brain in a Rodent Model of Inflammation Induced by Lipopolysaccharide in Male Rats.

Inflammation is considered to be a link between diabetes and central nervous system (CNS) disorders, including depression and anxiety. Metformin is suggested to have antioxidant, anti-inflammatory, and mood-improving effects. The aim of the current research was to investigate the effects of the antidiabetic drug metformin on depressive- and anxiety- like behaviors and oxidative stress in the brain in a rodent model of inflammation induced by lipopolysaccharide (LPS) in male rats. The rats were treated as follows: (1) Vehicle instead of metformin and lipopolysaccharide, (2) Lipopolysaccharide (1 mg/ kg) + vehicle instead of metformin, (3-5) Lipopolysaccharide + 50, 100, or 150 mg/ kg of metformin. After the behavioral tests, including open field (OF), elevated pulse maze (EPM), and force swimming (FS) tests, the brains were removed, and malondialdehyde (MDA), nitric oxide (NO) metabolites, total thiol, catalase (CAT) activity, interleukin-6 (IL-6) and superoxide dismutase (SOD) activity were determined. In the EPM, metformin increased the open arm time and entry and decreased closed arm time and entry. In the FS test, metformin lowered the immobility and increased active time compared to lipopolysaccharide. In the OF test, metformin increased total crossing and total distance, time spent, traveled distance, and crossing number in the central zone. As a result of metformin administration, IL-6, MDA, and NO metabolites were decreased while thiol content, SOD, and CAT activity were increased. The results indicated that the well-known antidiabetic drug metformin attenuated depressive- and anxiety-like behaviors induced by inflammation in rats. These beneficial effects are suggested to be due to their attenuating effects on neuroinflammation, oxidative stress, and NO in the brain.

In Vivo Chemosuppressive Effects of Kolaviron on 7,12-Dimethylbenzanthracene-Induced Mammary Lesions are Associated with Changes in Levels of Estrogen Receptor-α, CYP 1A1, Proinflammatory Cytokines, and Alterations to Metabolic Pathways Implicated in Mammary Carcinogenesis.

Garcinia kola is a medicinal food commonly consumed in Sub-Sahara Africa, for which Kolaviron (KV) is the active portion. As a follow-up to our earlier chemopreventive studies, we investigated the chemotherapeutic effects of KV on experimentally induced mammary carcinogenesis in female Wistar rats. Mammary carcinogenesis was induced using 80 mg/kg of 7,12-dimethylbenzanthracene (DMBA) administered by oral gavage. One hundred-fifty days post-DMBA induction, estrogen receptor-α (ER-α) levels were determined in the experimental rats before treatment with KV commenced. Treatment was done using 50, 100, and 200 mg/kg KV thrice a week for 4 weeks, after which the experiment was terminated. Significantly higher levels of estrogen receptor-α, CYP 1A1, malondialdehyde, formation of lobular neoplastic cells, epithelial hyperplasia, lymphocyte infiltration, and increased cytokine (interleukin-6 and tumor necrosis factor-α) activity were observed in DMBA-induced rats, which were attenuated in KV-treated rats. Tyrosine metabolism was exclusively enriched in DMBA-induced rats in contrast to KV-treated rats. Collectively, the results point to the chemotherapeutic potential of KV.

Exercise-induced changes in left ventricular strain are affected by interleukin-6 activity: An exploratory analysis of a randomised-controlled trial in humans with abdominal obesity.

Whilst the exercise-induced myokine interleukin-6 (IL-6) plays a beneficial role in cardiac structural adaptations, its influence on exercise-induced functional cardiac outcomes remains unknown. We hypothesised that IL-6 activity is required for exercise-induced improvements in left ventricular global longitudinal strain (LV GLS). In an exploratory study 52 individuals with abdominal obesity were randomised to 12weeks' high-intensity exercise or no exercise in combination with IL-6 receptor inhibition (IL-6i) or placebo. LV strain and volume measurements were assessed by cardiac magnetic resonance. Exercise improved LV GLS by -5.4% [95% CI: -9.1% to -1.6%] (P=0.007). Comparing the change from baseline in LV GLS in the exercise + placebo group (-4.8% [95% CI: -7.4% to -2.2%]; P<0.0004) to the exercise + IL-6i group (-1.1% [95% CI: -3.8% to 1.6%]; P=0.42), the exercise + placebo group changed -3.7% [95% CI: -7.4% to -0.02%] (P=0.049) more than the exercise + IL6i group. However, the interaction effect between exercise and IL-6i was insignificant (4.5% [95% CI: -0.8% to 9.9%]; P=0.09). Similarly, the exercise + placebo group improved LV global circumferential strain by -3.1% [95% CI: -6.0% to -0.1%] (P=0.04) more compared to the exercise + IL-6i group, yet we found an insignificant interaction between exercise and IL-6i (4.2% [95% CI: -1.8% to 10.3%]; P=0.16). There was no effect of IL-6i on exercise-induced changes to volume rates. This study underscores the importance of IL-6 in improving LV GLS in individuals with abdominal obesity suggesting a role for IL-6 in cardiac functional exercise adaptations.

Possible beneficial effects of nano chitosan against doxycycline toxicity in Nile tilapia (Oreochromis niloticus)

In aquaculture, improper and illogical antibiotic use could yield negative outcomes. Therefore, this study investigates the ameliorative effects of chitosan (CS) and chitosan nanoparticles (CSNPs) against the toxicity of the antibiotic doxycycline in Nile tilapia fish. The fish were divided into four distinct categories: the first group served as the control, the second group was subjected to doxycycline (40 mg/L), the third group experienced doxycycline (40 mg/L) and CS (0.5% in fish diet), and the fourth group was exposed to doxycycline (40 mg/L) with CSNPs (0.5% in fish diet) for 15 days. Levels of creatinine, uric acid, and aspartate aminotransferase activities notably elevated (p < 0.05) in the doxycycline-treated group when compared with the control group. Meanwhile, fish in the doxycycline-exposed group demonstrated significantly lower (p < 0.05) levels of red blood cells, hemoglobin, packed cell volume, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and neutrophils. Conversely, mean corpuscular volume, platelets, white blood cells, and lymphocytes exhibited notably higher (p < 0.05) levels in the doxycycline group. Other hematological indicators, including monocytes and eosinophils, showed no notable variances (p > 0.05) among the studied groups. Fish treated with doxycycline exhibited notably higher (p < 0.05) interleukin-1β and interleukin-6 activity in comparison to the control group. Additionally, histopathological changes were detected in the gills, liver, and kidney tissues of doxycycline-exposed fish when contrasted with the control group. Co-exposure of CS and CSNPs significantly restored most hematological, biochemical, and immunological parameters, as well as histopathological indices, in the fish, aligning these values closer to those observed in the control group in comparison to the fish treated solely with doxycycline. In conclusion, both CS and CSNPs played an important role in moderating the negative effects of doxycycline on fish, through improving hematological indices, reductions in creatinine, uric acid, and aspartate aminotransferase activity, as well as anti-inflammatory action through reductions in interleukin-1β and interleukin-6 activity.

Cinnamic Acid Ameliorates Acetic Acid-induced Inflammatory Response through Inhibition of TLR-4 in Colitis Rat Model.

Cinnamic acid, an active compound in cinnamon spp., has anti-inflammatory and antioxidant characteristics and is favorable in managing inflammatory bowel diseases. Evaluate cinnamic acid's effects on colitis in rats. To induce colitis in experimental rats, excluding the sham group, a 4% intrarectal solution of acetic acid was administered. The rats were then given oral doses of cinnamic acid at 30, 45, and 90 mg/kg for two days. The animals were assessed for macroscopic and microscopic changes, and the levels of inflammatory mediators such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and myeloperoxidase (MPO) were measured using Eliza kits. Additionally, real-time PCR was performed to examine the gene level of toll-like receptor 4 (TLR-4) in the colon. Effective reduction of inflammation in acetic acid-induced colitis was achieved through Cinnamic acid administration at doses of 45 and 90 mg/kg. The decrease was achieved by inhibiting the activities of TNF-α, IL-6, and MPO while downregulating the expression of TLR-4. It is important to note that macroscopic and microscopic evaluations were significant in determining the effectiveness of cinnamic acid in reducing inflammation. Downregulation of inflammatory cytokines and TLR-4 expression may contribute to cinnamic acid's anti-inflammatory effect.

Natural products as IL-6 inhibitors for inflammatory diseases: Synthetic and SAR perspective.

Interleukin-6 (IL-6), a pleiotropic cytokine, plays a pivotal role in the pathophysiology of various diseases including diabetes, atherosclerosis, Alzheimer's disease, multiple myeloma, rheumatoid arthritis, and prostate cancer. The signaling pathways associated with IL-6 offer promising targets for therapeutic interventions in inflammatory diseases and IL-6-dependent tumors. Although certain anti-IL-6 monoclonal antibodies are currently employed clinically, their usage is hampered by drawbacks such as high cost and potential immunogenicity, limiting their application. Thus, the imperative arises to develop novel small non-peptide molecules acting as IL-6 inhibitors. Various natural products derived from diverse sources have been investigated for their potential to inhibit IL-6 activity. Nevertheless, these natural products remain inadequately explored in terms of their structure-activity relationships. In response, our review aims to provide syntheses and structure activity perspective of natural IL-6 inhibitors. The comprehensive amalgamation of information presented in this review holds the potential to serve as a foundation for forthcoming research endeavors by medicinal chemists, facilitating the design of innovative IL-6 inhibitors to address the complexities of inflammatory diseases.

Interleukin-6 (IL-6) as a Marker of Endothelial Dysfunction Confirmed Using Flow Mediated Dilatation in Active Smoker

Background: The pro-inflammatory cytokine interleukin-6 (IL-6) is thought to be the catalyst for endothelial dysfunction caused by smoking. Flow-mediated dilatation (FMD) is a non-invasive method to identify the development of blood vessel endothelial dysfunction at an early stage. However, several studies have not sufficiently explained how smoking and IL-6 interact. Objective: This study focuses on the role of Interleukin-6 (IL-6) in the process of endothelial dysfunction confirmed using non-invasive modalities such as Flow Mediated Dilatation (FMD) in smokers related to the risk of cardiovascular disease in the future with the hope of providing insight in education to smokers to quit smoking. Material and Methods: In this study, we gathered 116 male participants, of which 56 were nonsmokers, and 60 were smokers, and we used IL-6 and FMD testing. SPSS for Windows version 23 will be used to analyze the data. Result: According to the analysis of laboratory tests, the average IL-6 level in light smokers was 67.7±3.2 and in moderate smokers it was 95.111.0; with a p-value of 0.009, indicating a significant difference among the groups. The average FMD in the smoking group was 5.4±0.4% (p-value 0.000), whereas the average FMD in the non-smoker samples was 10.9±0.9%. We also looked at the average FMD among light and moderate smokers based on Brinkman Index, which was 5.5±0.4% and 5.00.4%, with a p-value of 0.780, indicating no significant difference between the groups Conclusion: Compared to non-smokers, this study demonstrates a correlation between IL-6 and smoking activity. However, there was no statistically significant difference between the prevalence of endothelial dysfunction and the degree of smoking dependence in this investigation.

Serum Interleukin-6 in Schizophrenia: Associations with Clinical and Sociodemographic Characteristics.

Recently a significant part of schizophrenia studies have been focused on the role of cytokines, especially interleukin-6 (IL-6). Some authors have suggested a pathogenetic role for IL-6 in schizophrenia and concluded that therapy that centers on suppressing IL-6 activity may prove beneficial for certain categories of patients with the disorder. However, many questions about whether the changes in IL-6 levels in schizophrenia are primary, related to symptoms or caused by therapy, are concomitant metabolic disorders, are related to smoking or other secondary factors remain unanswered. To assess the level of serum IL-6 in patients with schizophrenia in comparison with healthy controls, as well as to study its association with clinical and socio-demographic characteristics. Some 125 patients with schizophrenia and 95 healthy volunteers were examined. The evaluation of IL-6 was performed by enzyme immunoassay. All patients were assessed using standardized psychometric instruments. Information from patient medical records on the course of the disease and treatment was analyzed. The level of IL-6 was significantly higher in the patients than in the healthy volunteers (z=2.58; p=0.0099), but among men the difference between the patients and volunteers was not significant. Statistically significant correlations were found between the level of serum IL-6 and the severity of the cognitive impairment of patients: (auditory [ρ=-0.31; p=0.00063] and working memory [ρ=-0.25; p=0.0065], hand-eye coordination [ρ=-0.29; p=0.0011], verbal fluency [ρ=-0.28; p=0.0019] and problem-solving capacity [ρ=-0.22; p=0.013]), total severity of schizophrenia symptoms (PANSS, ρ=0.22; p=0.016), PANSS positive subscale (ρ=0.18; p=0.048), and the age of manifestation (ρ=0.20; p=0.025) and disease duration (ρ=0.18; p=0.043). The level of IL-6 was the lowest in patients treated with third-generation antipsychotics, and the highest in those treated with first-generation antipsychotics (H=6.36; p=0.042). Moreover, in hospital patients, the level of IL-6 was significantly higher than in outpatients and inpatients hospitals (H=18.59; p=0.0001). The study confirmed that there are associations between the serum IL-6 level and schizophrenia, the age of the patient, duration of the disease and how late in one's life cycle it began manifesting itself, as well as a number of clinical characteristics. Considering that IL-6 is associated with a wide range of symptoms that are loosely controlled by antipsychotics, this biochemical marker needs to be studied to look into how closely its level tracks with an unfavorable course of schizophrenia. That would require further prospective studies.

Interleukin-6 and disease activity in childhood systemic lupus erythematosus

Background Systemic lupus erythematosus (SLE) is a complex disease with various manifestations. Interleukin-6 (IL-6) is a pleiotropic cytokine with a wide range of biological activities which plays an important role in immune regulation and inflammation. Serum level of IL-6 may be used as a parameter of disease activity, especially in pediatric SLE patients with mild disease activity or total remission with conflicting clinical manifestations and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) scores.&#x0D; Objective To identify the characteristics of serum IL-6 concentration in pediatric SLE with mild disease activities and total remission.&#x0D; Methods This case-control study was performed at the allergy-immunology outpatient clinic, Department of Child Health Dr. Cipto Mangunkusumo Hospital, Jakarta and Dr. Sardjito Hospital, Yogyakarta. Serum IL-6 concentration and disease activity were assessed in all pediatric SLE patients aged 1-18 years old. Disease activity was assessed with SLEDAI scores and serum level of IL-6 was measured by enzyme-linked immunosorbent assay.&#x0D; Results Among 60 subjects included in this study, 30 subjects with mild activities were in the case group and 30 subjects with total remissions were in the control group. There was no difference in serum IL-6 concentration between the case and control group (OR 0.483; 95%CI 0.041to 5.628; P=0.500). In this study, 2 subjects with urinary tract infection had high serum IL-6 concentrations.&#x0D; Conclusion There is no difference in serum IL-6 concentration between pediatric SLE patients with mild disease activities compared to total remissions.

Coexistence of Granuloma Annulare and Castleman’s Disease: A Case Report

Granuloma annulare (GA) is a self-limited disorder that may present as localized or general. Localized GA, the most common form, presents as an erythematous annular plaque without scale. The generalized form of GA presents as widespread erythematous papules and plaques over the trunk and extremities. A cross-sectional study in the United States found a female to male prevalence ratio of 3:1 with a higher prevalence in the fifth decade of life.1 The etiology of GA is largely unknown; however, a number of potential inciting factors have been reported including viral infections, drug exposure, trauma, and insect bites.&#x0D; Castleman’s Disease is a rare heterogenous lymphoproliferative disorder that encompasses a spectrum of clinicopathologic manifestations. Unicentric disease affects one lymph node or chain of lymph nodes with primarily hyaline vascular changes. The multicentric form affects multiple chains of lymph nodes with primarily plasmacytic changes. The pathogenesis is related to hypercytokinemia due to dysregulated interleukin-6 (IL-6) activity in unicentric disease and dysregulated IL-6 and human herpes virus-8 in multicentric disease.&#x0D; We report a case of concomitant granuloma annulare and Castleman’s Disease.

Differential response to interleukin-1 blockade with anakinra on cardiorespiratory fitness in patients with heart failure with preserved ejection fraction stratified according to ejection fraction

Abstract Background Patients with heart failure with preserved ejection fraction (HFpEF) are a heterogeneous clinical population and might respond differently to pharmacologic therapies based on their baseline left ventricular EF (LVEF) (50-59% vs. ≥60%). Interleukin-1 (IL-1) is a key pro-inflammatory cytokine being explored as a therapeutic target in HFpEF for its effects on numerous outcomes including cardiorespiratory fitness (CRF). Purpose Our aim was to examine changes in CRF and C-reactive protein (CRP), a surrogate for IL-1 activity, in patients with HFpEF treated with the IL-1 recombinant human receptor antagonist anakinra, stratified according to resting LVEF. Methods We analyzed data from 32 patients (median age 55 [51-64] years, 24 (75%) female) with HFpEF, defined as a presence of HF symptoms, LVEF≥50%, and evidence of left ventricular diastolic dysfunction or elevated filling pressures. All patients were treated with anakinra 100 mg subcutaneously daily for 12 [2-12] weeks. Patients were stratified into two groups: LVEF 50-59% and LVEF ≥60%. At baseline and during treatment, patients underwent cardiopulmonary exercise testing and venipuncture to measure changes in peak oxygen consumption (peak VO2) and CRP. Categorical and continuous variables are expressed as N (%) and median (interquartile range) and were compared with the chi-square and Mann-Whitney test, respectively. The within-group paired differences were compared using the Wilcoxon signed-rank test. Results The cohort included 15 patients with LVEF 50-59% and 17 with LVEF ≥60%, without significant differences in age and sex between groups. In patients with LVEF 50-59% and ≥60%, baseline peak VO2 was 14.1 [13.1–17.3] and 13.8 [11.8-18.6] mLO2·kg-1·min-1, respectively (p = 0.82), and CRP was 6.2 [3.4-15.9] and 6.1 [3.7-17.6] mg/L, respectively (p = 0.74). A significant increase in peak VO2 (from 13.8 [11.8-18.6] to 15.8 [12.7-19.9] mLO2·kg-1·min-1, p = 0.033) and decrease in CRP (from 6.2 [3.4-15.9] mg/L to 3.4 [1.0-6.7] mg/L, p = 0.003) was seen in patients with LVEF ≥60%; whereas in those with LVEF 50-59% there were no significant changes in peak VO2 (from 14.1 [13.1–17.3] to 14.1 [11.0–17.3] mLO2·kg-1·min-1, p = 0.25) despite a significant reduction in CRP (from 6.1 [3.7-17.6] mg/L to 1.4 [0.8-11.4] mg/L, p = 0.016). We found a difference in the interval changes in peak VO2 (0 [from -1.1 to +0.3] vs +1.7 [from -0.7 to +2.3] mLO2·kg-1·min-1 in patients with LVEF 50-59% and ≥60%, respectively, p = 0.01), without differences in the interval change in CRP levels (–3.5 [from -6 to -2.1] vs –3.0 [from -11.4 to -2.1] mg/L, respectively, p = 0.91). Figure 1. Conclusions Patients with HFpEF demonstrate differential peak VO2 response to anakinra treatment based on resting LVEF, with those with a LVEF ≥60% showing greater improvement in CRF. Further randomized control trials are needed to validate these data and explore the potential underlying mechanisms.

AI-driven Discovery of Celecoxib and Dexamethasone for Exploring their Mode of Action as Human Interleukin (IL-6) Inhibitors to Treat COVID-19-induced Cytokine Storm in Humans.

In the case of COVID-19 patients, it has been observed that the immune system of the infected person exhibits an extreme inflammatory response known as cytokine release syndrome (CRS) where the inflammatory cytokines are swiftly produced in quite large amounts in response to infective stimuli. Numerous case studies of COVID-19 patients with severe symptoms have documented the presence of higher plasma concentrations of human interleukin-6 (IL-6), which suggests that IL-6 is a crucial factor in the pathophysiology of the disease. In order to prevent CRS in COVID-19 patients, the drugs that can exhibit binding interactions with IL-6 and block the signaling pathways to decrease the IL-6 activity may be repurposed. This research work focused on molecular docking-based screening of the drugs celecoxib (CXB) and dexamethasone (DME) to explore their potential to interact with the binding sites of IL-6 protein and reduce the hyper-activation of IL-6 in the infected personnel. Both of the drugs were observed to bind with the IL-6 (IL-6 receptor alpha chain) and IL-6Rα receptor with the respective affinities of -7.3 kcal/mol and -6.3 kcal/mol, respectively, for CXB and DME. Moreover, various types of binding interactions of the drugs with the target proteins were also observed in the docking studies. The dynamic behaviors of IL-6/IL-6Rα in complex with the drugs were also explored through molecular dynamics simulation analysis. The results indicated significant stabilities of the acquired drug-protein complexes up to 100 ns. The findings of this study have suggested the potential of the drugs studied to be utilized as antagonists for countering CRS in COVID-19 ailment. This study presents the studied drugs as promising candidates both for the clinical and pre-clinical treatment of COVID-19.

In vitro response of dental pulp stem cells to dural substitute grafts: Analysis of cytocompatibility and bioactivity.

The objective of this study was to assess and compare the cytocompatibility of decellularized porcine small intestine submucosal dural graft from Biodesign (BD) and polyester urethane-based Neuro-Patch (NP) dural substitute with the mineral trioxide aggregate (MTA) and the cyanoacrylate-based Histoacryl surgical adhesive. Furthermore, the study evaluated the inflammatory response and osteogenic differentiation of human dental pulp stem cells (hDPSCs) when cultured in direct contact with the dural substitutes in comparison with MTA. The viability of hDPSCs in direct contact with the tested materials was investigated in vitro by a CCK-8 assay. Additionally, the effects of dural substitutes and MTA on the expression of the inflammatory mediator interleukin-6 (IL-6) was investigated via enzyme-linked immunosorbent assay (ELISA), whilst effects on the differentiation were evaluated using alizarin red staining, alkaline phosphatase staining, ELISA and energy-dispersive X-ray elemental mapping. The dural substitutes were cytocompatible and promoted cellular adhesion. The Histoacryl and MTA demonstrated cytotoxicity in fresh preparations but showed a more favourable cellular reaction when set. Investigations of biological activity indicated that dural substitute membranes did not induce an inflammatory response or osteogenic differentiation of hDPSCs. In contrast, MTA induced the expression of IL-6 and alkaline phosphatase activity contributing to enhanced differentiation and mineralization. The dural substitute membranes showed cytocompatibility, did not provoke an inflammatory response and maintained the stemness of hDPSCs better than MTA. Additionally, the set Histoacryl surgical adhesive demonstrated good biocompatibility. Taken together, these results highlight the potential use of dural substitutes in regenerative endodontic procedures as coronal barriers alternative to MTA to reduce the incidence of intracanal calcifications.

Network Pharmacology, Molecular Docking, Molecular Dynamics Simulation, and in vitro Experiments to Explore the Mechanism of Asiatic Acid Inhibiting Acetaldehyde-Induced Activation of Hepatic Stellate Cells

Purpose: This study utilized network pharmacology, molecular docking, molecular dynamics simulation, and in vitro experimental verification to explore the mechanisms of action by which asiatic acid (AA) inhibits acetaldehyde-induced activation of hepatic stellate cells (HSCs). Methods: Databases were screened to identify intersections between AA and alcoholic liver fibrosis. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) Enrichment analysis were performed, followed by molecular docking and molecular dynamics simulation to predict further the interactions between AA and cross-targets. Mechanisms of action of AA were assessed experimentally in HSC-T6 rat HSCs. Results: Screening of the relevant databases using web pharmacology identified several genes, including those encoding signal transducer and activator of transcription 3, nuclear factor kappa-B (NF-κB)-P65 (RELA), tumor necrosis factor α (TNF-α), interleukin 6 (IL-6), and interleukin 1 beta (IL-1β), that were closely associated with alcoholic liver fibrosis. GO and KEGG enrichment analysis showed that the alcoholic liver disease pathway was the most relevant one, and molecular docking showed critical binding activity of RELA, TNF-α, IL-6, and IL-1β with AA. Molecular dynamics simulation showed that the binding between RELA and AA was stable. AA at concentrations of 0 to 30 μM were determined to be nontoxic to HSC-T6 cells. Real-time quantitative polymerase chain reaction and Western blotting showed that the levels of expression of α-SMA and type 1 collagen were higher in acetaldehyde-treated than untreated HSC-T6 cells, with AA reducing their expression dose-dependently in acetaldehyde-treated cells. Western blotting also showed that AA could upregulate the expression of the apoptotic proteins BCL2 associated X and cleaved caspase 3 and could inhibit the phosphorylation of RELA and IKB-α. Conclusion: AA inhibited acetaldehyde-induced HSC-T6 cell proliferation and suppressed the secretion of fibrillar factors by inhibiting RELA phosphorylation.

Effect of Diabetic Neuropathy on Reparative Ability and Immune Response System.

The effects of diabetes can be divided into short, medium and long term and various human organ systems can be effected. The present study aimed to determine how much the duration of diabetes mellitus (DM) affect the reparative ability of the body, immune response and the development of DM complications. Interleukin 1-β (IL-1β) and Interleukin 6 (IL-6) were monitored as specific indicators of inflammatory reaction and C-reactive protein (CRP), leukocyte count (WBC) and sedimentation rate (ESR) as general markers of inflammatory reaction. Tumour necrosis factor α (TNF-α) and transforming growth factor β1 (TGF-β1) were observed as indicators of reparative ability and polyneuropathy. All interleukins were determined by ELISA and evaluated spectrophotometrically. Michigan Neuropathy Screening Instrument (MNSI) is performed for neuropathy examination. Patients with diabetes mellitus were divided into 3 groups, according to duration of diabetes mellitus. IL-6 levels correlated with clinical stage of diabetic polyneuropathy at p = 0.025 R = 0.402; with CRP at p = 0.0001, R = 0.784 as well as correlation of CRP and MNSI score (R = 0.500, p = 0.034) in a group of patients with DM lasting up to 10years. The reparative ability of the body is reduced by physiological age and ages of DM duration. The immune response is weakened in DM additionally. The dual activity of cytokines IL-6 and TGF-β1 is present in long-duration Diabetes Mellitus.

CHANGES IN MARKERS OF BONE TISSUE REMODELING AND THE INFLAMMATORY PROCESS IN THE BLOOD SERUM OF WHITE RATS IN CASE OF DEFECT FILLING OF THE FEMUR WITH IMPLANTS BASED ON POLYLACTIDE AND TRICALCIUMPHOSPHATE WITH MESENCHYMAL STEM CELLS

Objective. Based on the analysis of markers of inflammation and metabolism of bone tissue in the blood serum of laboratory rats, to evaluate the course of bone remodeling after filling the defect in the distal metaphysis of the femur with 3D-printed implants based on polylactide and tricalcium phosphate (3D-I) alone or in combination with mesenchymal stromal cells (MSCs). Methods. 53 white rats were used, which were divided into groups: intact (5 animals) — the operation was not performed; Control (15) — 3D-I; Experiment I (15) — 3D-I + cultured alloMSCs; Experiment II (15) — 3D-I + introduction of alloMSCs into the area of surgical intervention 7 days after implantation. The following were studied: the content of glycoproteins (GP), interleukin-6 (IL-6), osteocalcin, chondroitin sulfates (CS), total protein, calcium, alkaline (AlP) and acid phosphatase (AP) activity, and their ratio, mineralization indices were calculated. Results. Compared with intact animals, higher indicators were determined in the rats of the Control group: the content of GP by 39.73; 32.88; 23.29 %; CS — 250.00; 222.09 and 196.51 %, AlP activity — 81.67, 51.03, 39.36 %, on the 15th, 30th, and 90th days of the experiment; IL-6 — 44.89; 60.06 % on the 15th and 30th days. In the rats of the Experiment I g roup: the content of GP — by 82.19; 65.75, 57.53 %, IL-6 — 72.14; 96.59; 79.88 %, CS — 306.98; 276.74; 253.49 %; AlP activity — 63.73; 129.70; 51.28 %, on the 15th, 30th and 90th days of the experiment. In the Experiment II group: on the 15th, 30th and 90th days, the content of GP was higher by 27.40; 26.03; 129.18 %; CS — by 175.58; 137.21 and 115.12 %; AlP activity — 192.99; 178.02, 76.31 %; on the 15th and 30th days: IL-6 — by 37.46; 20.74 %. Conclusions. In the case of filling the defect with 3D-printed implants, biochemical signs of moderate inflammation were determined; 3D-printed implants together with MSCs — pronounced inflammation, slowing of bone formation, formation of connective tissue; 3D-printed implants with postoperative injection of MSCs — moderate inflammation and optimal conditions for healing the defect with bone tissue.