Anti-inflammatory Cytokine Genes Research Articles

Effect of rifampicin on Toll-like receptor system gene expression in brain structures of rats with prenatal alcohol exposure

Alcohol intake during pregnancy may affect the normal course of fetal development, causing the symptoms of fetal alcohol spectrum disorders (FASD). There is evidence that a number of prenatal pathologic conditions exhibit altered expression of several pro- and anti-inflammatory cytokines. In our study, we focused on studying expression of numerous pro- and anti-inflammatory cytokine genes in the forebrain and temporal regions of rat brain during the postnatal development, thus modeling prenatal alcohol exposure effects (PAE). We also evaluated expression of genes associated with regulation of genes controlling expression of pro- and anti-inflammatory cytokines. Moreover, the objectives of our study included pharmacological correction of the observed changes by rifampicin (Rif), a potential pharmacological agent which had a neuroprotective effect shown by other studies,. The experimental model of PAE was produced by oral intake of 15% ethanol solution to pregnant female rats throughout pregnancy. The drug injections were performed to the pups from the 1 to the 7 postnatal days. Brain structures were sampled for gene expression analysis on the 8 postnatal day. The results of the study showed distinct changes in Tlr3 and Tlr4 gene expression in anterior and temporal lobes of brain on the 8 day of postnatal development. Expression of Myd88 and Ticam genes showed multidirectional changes among the studied brain structures of PAE rats. The increased mRNA level of proinflammatory genes was noted. Usage of Rif in experiments showed the ability of Rif (50 mg/kg) to correct the observed long-term pathological changes in the expression of the genes under study. It is of interest to study the dose-dependent effect in the future, as well as to investigate the revealed changes at the level of protein analysis. In future studies, it seems important to evaluate TLR signaling system in other brain structures with PAE, as well as at different terms of postnatal development in ontogenesis.

Sleeve gastrectomy and pancreatic omentoplasty decrease insulin resistance: Impact on pro-inflammatory and anti-inflammatory cytokine gene expression

Sleeve gastrectomy and pancreatic omentoplasty decrease insulin resistance: Impact on pro-inflammatory and anti-inflammatory cytokine gene expression

Dietary Solid-state-fermentation product of Bacillus velezensis T23 alleviate hepatic steatosis, oxidative stress, gut barrier damage, and microbiota dysbiosis in juvenile genetically improved farmed tilapia (GIFT, Oreochromis niloticus)

The application of commensal probiotic bacteria is one of the preferable green feed additives for antibiotic substitution in aquaculture, while the application methods and detailed action mechanisms of commensal bacteria isolated from aquatic animals are not consistent with normal mammalian-derived probiotics. The purpose of this study was to evaluate the effects of solid-state fermentation product of autochthonous Bacillus velezensis T23 on improving hepatic steatosis, liver antioxidation capacity, gut injury and gut microbiota profile of genetically improved farmed tilapia (GIFT, Oreochromis niloticus). The T23 probiotic was added to the basal diet at a level of 0.00, 0.05, 0.1, 0.2 and 0.3 g/kg to develop five experimental diets, respectively. After 4 weeks of the feeding trial, the results showed that T23 supplementation notably reduced the hepatocytes vacuolization and the content of liver TAGs by downregulated the mRNA expression of the lipogenesis gene (fas), while upregulated the mRNA expression of lipid degradation genes (cpt1 and pparα) (p < 0.05). Compared with the control, the 0.3T23 group showed significantly upregulated expression levels of anti-inflammatory cytokine genes (tgf-β and il-10) in the liver of the fish (p < 0.05). Besides, the activities of antioxidase SOD and CAT were significantly upregulated in T23 treatment groups, compared with the control (p < 0.05). Fish groups fed with 0.2 g/kg and 0.3 g/kg T23 supplemented diet also exhibited significantly lower intestinal injury and intestinal inflammation via upregulating the expression of tight junction protein gene claudin, anti-inflammatory factor gene il-10 (p < 0.05). However, markedly downregulation of the mRNA level of pro-inflammatory cytokine il-1β (p < 0.05) was observed in 0.05 and 0.3T23 groups, compared with the control. 16S rRNA sequencing analysis data on gut microbiota indicated that supplementation of T23 at lower levels (0.05, 0.1 and 0.2) resulted in a remarkable elevation of the abundance of phylum Firmicutes and reduced the abundance of Proteobacteria, and the ratio of (Firmicutes +Bacteroidota +Fusobacteria) /Proteobacteria. On the contrary, feeding with 0.3T23 negatively affects the gut microbiota diversity and structure of tilapia. Therefore, a 0.2 g/kg dose is suggested to supplement the diet of Nile tilapia to improve liver health and modulate the gut microbiota profile of the fish positively. Overall, these findings provide a shred of beneficial evidence of the application of B. velezensis T23 as a green feed additive to improve the productivity and profitability of tilapia farming.

Interventions to modify the habituation of biological responses to repeated stress in healthy adults: a randomized controlled trial

BackgroundChanges in response patterns of biological stress systems, including responses of the sympathetic nervous system (SNS) and the hypothalamus–pituitary–adrenal (HPA) axis to repeated stress, can promote the development and progression of chronic diseases via changes in downstream inflammatory processes. The aim of this project is thus to investigate, whether habituation of biological stress system activity including responses of the inflammatory system can be modified. Aiming to test for possible paths of action, a randomized controlled study with two intervention programs designed to manipulate cognitive coping strategies will be carried out. By increasing either ruminative or self-compassionate thoughts among healthy young adults, the intervention programs are expected to affect the regulation of occurring emotions as expressed by the responsiveness of biological systems during repeated stress exposure.MethodsIn this study, a total of 120 healthy adults will complete the Trier Social Stress Test (TSST) on two consecutive days. Immediately after the first stress induction, participants will be randomly assigned to two experimental conditions designed to manipulate cognitive coping strategies (rumination vs. self-compassion) or a control condition. Measures of HPA axis (salivary cortisol) and autonomic activity (salivary alpha amylase, heart rate, heart rate variability) as well as inflammatory markers (plasma interleukin(IL)-6, expression rates of pro- and anti-inflammatory cytokine genes) will be repeatedly assessed throughout the experimental sessions. Response and habituation indices of these measures will be calculated and compared between the experimental conditions and the control condition.DiscussionThe results should provide insight into whether modifying response patterns of biological stress systems could reverse a significant biological mechanism in the development of stress-related diseases.Trial registrationGerman Clinical Trials Register (DKRS), DRKS00034790. Registered on August 12, 2024, https://www.drks.de/DRKS00034790.

MicroRNA-30a-5p as a target for pharmacological correction of pathological conditions of the nervous system

In the brain, the main inducers of neuroinflammation are proinflammatory cytokines, chemokines, reactive oxygen species and other mediators produced by microglia, astrocytes and endothelial cells. Chronic neuroinflammatory conditions are manifested by the infiltration of peripheral immune cells through the blood-brain barrier and cause tissue damage in the central nervous system, promoting glial activation and increasing the permeability of the blood-brain barrier. According to a number of studies, one of the regulators of these processes is small non-coding RNA, or microRNA, which can either contribute to disease progression or, conversely, reflect an attempt by the nervous system to prevent excessive damage and restore homeostasis. Studying the role of microRNA. miR-30a-5p among others, in these processes can shed light on the pathogenetic mechanisms underlying a number of neurological diseases and lead to the discovery of new therapeutic agents. In this review, we discuss the role of miR-30a-5p in the regulation of pro- and anti-inflammatory cytokine gene expression, possible mechanisms of its action, and the use of miR-30a-5p as a potential therapeutic target for pharmacological correction of neuroinflammation in pathological conditions of the nervous system.

The pro-inflammatory responses of innate immune cells to Leishmania RNA virus 2-infected L. major support the survival and proliferation of the parasites

Infection of Leishmania by Leishmania RNA virus (LRV) has been proposed as a pathogenic factor that induces pro-inflammatory responses through the TLR3/TLR4 signaling pathway. We investigated the effect of L. major infection by LRV2 on innate immune cell responses (human neutrophil (HL-60) and macrophage (THP-1) cell lines). The expression levels of pro- and anti-inflammatory cytokine and chemokine genes as well as genes involved in the amino acid metabolism of arginine were then investigated by RT-qPCR. Moreover, the expression of TLR genes and their downstream signaling pathways were compared in THP-1 cells infected with the two isolates. Apoptosis was also evaluated in infected THP-1 and HL-60 cells using the PI/Annexin V flow cytometry assay. In both cell lines, the expression of pro-inflammatory cytokines increased in response to LRV2+ L. major (Lm+), and the expression of chemokines shifted toward macrophage recruitment. In contrast to LRV2- L. major (Lm-), Lm + infected THP-1 cells acquired the M2-like phenotype. The presence of LRV2 increased the gene expression of TLRs and their signaling pathways, especially TLR3 and TLR4, which was proportional to the increase in pro-inflammatory cytokines. In addition, Lm + increased the expression of IL-10 and IFN-β, which contribute to the survival and growth of the parasite in the phagolysosome. Altogether, our results showed that Lm + could stimulate pro-inflammatory responses that promote parasite replication and stabilization in the host.

PPARβ/δ Agonist GW0742 Modulates Microglial and Astroglial Gene Expression in a Rat Model of Temporal Lobe Epilepsy.

The role of astroglial and microglial cells in the pathogenesis of epilepsy is currently under active investigation. It has been proposed that the activity of these cells may be regulated by the agonists of peroxisome proliferator-activated nuclear receptors (PPARs). This study investigated the effects of a seven-day treatment with the PPAR β/δ agonist GW0742 (Fitorine, 5 mg/kg/day) on the behavior and gene expression of the astroglial and microglial proteins involved in the regulation of epileptogenesis in the rat brain within a lithium-pilocarpine model of temporal lobe epilepsy (TLE). TLE resulted in decreased social and increased locomotor activity in the rats, increased expression of astro- and microglial activation marker genes (Gfap, Aif1), pro- and anti-inflammatory cytokine genes (Tnfa, Il1b, Il1rn), and altered expression of other microglial (Nlrp3, Arg1) and astroglial (Lcn2, S100a10) genes in the dorsal hippocampus and cerebral cortex. GW0742 attenuated, but did not completely block, some of these impairments. Specifically, the treatment affected Gfap gene expression in the dorsal hippocampus and Aif1 gene expression in the cortex. The GW0742 injections attenuated the TLE-specific enhancement of Nlrp3 and Il1rn gene expression in the cortex. These results suggest that GW0742 may affect the expression of some genes involved in the regulation of epileptogenesis.

Isolation and characterization of plant-derived exosome-like nanoparticles from Carica papaya L. fruit and their potential as anti-inflammatory agent.

Inflammation is an immune system response that identifies and eliminates foreign material. However, excessive and persistent inflammation could disrupt the healing process. Plant-derived exosome-like nanoparticles (PDENs) are a promising candidate for therapeutic application because they are safe, biodegradable and biocompatible. In this study, papaya PDENs were isolated by a PEG6000-based method and characterized by dynamic light scattering (DLS), transmission Electron Microscopy (TEM), bicinchoninic acid (BCA) assay method, GC-MS analysis, total phenolic content (TPC) analysis, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. For the in vitro test, we conducted internalization analysis, toxicity assessment, determination of nitrite concentration, and assessed the expression of inflammatory cytokine genes using qRT-PCR in RAW 264.7 cells. For the in vivo test, inflammation was induced by caudal fin amputation followed by analysis of macrophage and neutrophil migration in zebrafish (Danio rerio) larvae. The result showed that papaya PDENs can be well isolated using the optimized differential centrifugation method with the addition of 30 ppm pectolyase, 15% PEG, and 0.2 M NaCl, which exhibited cup-shaped and spherical morphological structure with an average diameter of 168.8±9.62 nm. The papaya PDENs storage is stable in aquabidest and 25 mM trehalose solution at -20˚C until the fourth week. TPC estimation of all papaya PDENs ages did not show a significant change, while the DPPH test exhibited a significant change in the second week. The major compounds contained in Papaya PDENs is 2,3-dihydro-3,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP). Papaya PDENs can be internalized and is non-cytotoxic to RAW 264.7 cells. Moreover, LPS-induced RAW 264.7 cells treated with papaya PDENs showed a decrease in NO production and downregulation mRNA expression of pro-inflammatory cytokine genes (IL-1B and IL-6) and an upregulation in mRNA expression of anti-inflammatory cytokine gene (IL-10). In addition, in vivo tests conducted on zebrafish treated with PDENs papaya showed inhibition of macrophage and neutrophil cell migration. These findings suggest that PDENs papaya possesses anti-inflammatory properties.

Hyperexpression of Proinflammatory Cytokines in Blood as a Biomarker of Systemic Inflammatory Response in Schizophrenia: Scoping Review

Introduction: An imbalance of the genetically determined cytokine response plays a key role in the etiology of treatment-resistant schizophrenia (TRS). In recent years, an attempt has been made to evaluate the prognostic role of systemic inflammation in the development of TRS. The problem requires a multidisciplinary approach on the part of the specialists in the following clinical disciplines: psychiatry, immunology, experimental medicine and pharmacogenetics. The solution of this problem is possible with the involvement of preventive and personalized medicine. The purpose: Evaluation the prognostic role of genetic polymorphisms of pro-inflammatory cytokines in the development of TRS. Materials and Methods: We conducted a keyword-based analysis of the English and Russian-language articles published within the past 5 years. The following databases were used in the study: PubMed, MedLine, Web of Science Core Collection (Clarivate Analytics), Web Science, Russian Science Citation Index, Scopus, Scientific Research, Google Scholar, Oxford Press, and eLibrary. Results: In a number of the analyzed works, an increased level of pro-inflammatory cytokine production was noted in patients with TRS. Based on this, single nucleotide variants (SNVs), their influence on the expression of pro- and anti-inflammatory cytokine genes, as well as their predictor role in the development of TRS. The most promising SNVs for further studies were identified. Conclusion: The risk of developing TRS is associated with a genetically determined status of the cytokine response and its regulation. Studies of the association of various SNVs of genes encoding pro-inflammatory cytokines in the Russian Federation need to be continued.

Proinflammatory cytokines IL-6, IL-1β, TNF-α in infective endocarditis

Aim. To study the features of macrophages in the tissues of resected valves in operated patients with infective endocarditis (IE), their significance and interaction with inflammatory markers to improve the effectiveness of IE diagnosis. Materials and methods. Prospectively the research included 25 adult patients with active IE (Duke criteria 2015) and 24 patients with heart defects without IE, hospitalized in a cardiosurgical hospital in Moscow (2021–2022). A standard laboratory and instrumental examination was carried out for the diagnosis of IE, including etiological diagnosis with microbiological and molecular biological methods, and echocardiographic examination of heart. Additionally, the neutrophil-to-lymphocyte ratio (NLR) was calculated. The study of macrophages was carried out in the tissues of resected valves with the determination of the expression of pro- and anti-inflammatory cytokine genes, macrophage markers (CD 68+) using real-time PCR. Results. Increased expression of proinflammatory cytokines IL-1β, TNF-α and IL-6 was revealed in the group of operated patients with IE with significant differences in IL-1β (CI [IQR] 0.00367 [0.00047–0.01553] vs 0.00018 [0.00012–0.00262]; p0.05) and IL-6 (CI [IQR] 0.00367 [0.00047–0.01553] vs 0.00018 [0.00012–0.00262]; p0.05) and IL-6 (CI [IQR] 0.00338 [0.00066–0.01674] vs 0.00054 [0.00044–0.00378]; p0.05). The expression of anti-inflammatory cytokines in valve tissues prevailed in the control group without significant differences from patients with IE. The macrophage marker CD 68+ was revealed in all examined patients with a significant quantitative predominance in the group of patients with IE. There were no differences in the expression of pro- and anti-inflammatory cytokines depending on the presence of embolic events, intracardiac complications, etiological affiliation to S. aureus, as well as hospital mortality and combined endpoint (death from all causes or recurrence of IE 6 months after surgery) in patients with IE with or without events. Cytokines IL-1β and IL-6 positively correlated with each other, with leukocytes and NLR. ROC analysis determined that IL-1β and NLR had the most favorable features for the diagnosis of IE [IL-1β AUC 0.816 (p=0.02), NLR AUC 0.807 (p=0.03)]. IL-6 did not show a diagnostic value in IE. The threshold value for IL-1β was 0.00029 (sensitivity 86.4%, specificity 60.0%, prognostic value of negative result 75.0% and positive 76.0%, AUC 0.761; p=0.008). Conclusion. The valve macrophages of patients with IE express elevated levels of proinflammatory cytokines IL-1β and IL-6, regardless of etiological affiliation or complicated course of IE. IL-1β has a high diagnostic value for determining the inflammatory activity in IE.

Polymorphism of IL4 and IL13 anti-inflammatory cytokine genes in indigenous populations of the russian Arctic

Living conditions in the Arctic zone are extremely unfavorable for living and contribute to the development of a number of diseases due to suppression of the functions of the immune system. Cytokines are one of the main mediators of the immune system; they are encoded by genes with a high degree of polymorphism. This study examined the distribution of genetic variants in the cytokine genes (rs2243250 IL4, rs1800925 IL13) among the Nenets, Dolgan-Nganasan and Slavs. It was previously established that these mutations are associated with the level of expression of these interleukins and their production, which leads to changes in the impairment of the immune response to the influence of the pathogen. The study showed the predominance of genotypes CT and TT of the rs2243250 IL4 polymorphism, and genotype CC of the rs1800925 IL13 polymorphism in the Nenets and Dolgan-Nganasan populations. The results obtained suggest that the indigenous inhabitants of the Russian Arctic have a genetically determined rapid immune response and protection against the development of allergic diseases compared to the Slavs.

Enterococcus durans 98D alters gut microbial composition and function to improve DSS-induced colitis in mice

Enterococcus durans, is a potential functional strain with the capacity to regulate intestinal health and ameliorate colonic inflammation. However, the strain requires further investigation regarding its safety profile and potential mechanisms of colitis improvement. In this study, the safety of E. durans 98D (Ed) as a potential probiotic was studied using in vitro methods. Additionally, a dextran sulfate sodium (DSS)-induced murine colitis model was employed to investigate its impact on the intestinal microbiota and colitis. In vitro antimicrobial assays revealed Ed sensitivity to common antibiotics and its inhibitory effect on the growth of Escherichia coli O157, Streptococcus pneumoniae CCUG 37328, and Staphylococcus aureus ATCC 25923. To elucidate the functional properties of Ed, 24 weight-matched 6-week-old female C57BL/6J mice were randomly divided into three groups (n = 8): NC group, Con group (DSS), and Ed group (DSS + Ed). Ed administration demonstrated a protective effect on colitis mice, as evidenced by improvements in body weight, colonic length, reduced disease activity index, histological scores, diminished splenomegaly, and decreased goblet cell loss. Furthermore, Ed downregulated the expression of the pro-inflammatory cytokine genes (IL-6, IL-1β, and TNF-α) and upregulated the expression of the anti-inflammatory cytokine gene IL-10. The 16S rRNA gene sequencing revealed significant alterations in microbial α-diversity, with principal coordinate analysis indicating distinct differences in microbial composition among the three groups. At the phylum level, the relative abundance of Actinomycetota significantly increased in the Ed-treated group. At the genus level, Ed treatment markedly elevated the relative abundance of Paraprevotella, Rikenellaceae_RC9, and Odoribacter in DSS-induced colitis mice. In conclusion, Ed exhibits potential as a safe and effective therapeutic agent for DSS-induced colitis by reshaping the colonic microbiota.

Extracellular vesicles secreted by the ТНР-1 cells influence on the inflammation gene expression in zebrafish

Extracellular vesicles secreted by immune cells may play a significant role in the initiation, maintenance, and progression of systemic inflammation. The aim of the study was to investigate the regulatory effect of extracellular vesicles (EVs) produced by activated monocyte-like THP-1 cells on expression levels of inflammatory genes in a zebrafish. Real-time PCR analysis was performed to investigate the relative expression levels of il-1β, il-6, tnf-α, ifn-γ, mpeg1.1, mpeg1.2, mpx, and il-10 genes in the brain, liver, and heart of zebrafish followed by intracelomic injection of EVs produced by THP-1 cells activated with tumor necrosis factor (TNF) and phorbol-12-myristate-13-acetate (PMA) at different concentrations. EVs, secreted by activated THP-1 cells with TNF at a concentration of 10 ng/mL and PMA at concentrations of 16 and 50 ng/mL, reduced the expression levels of il-1β, ifn-γ, tnf-α, mpx, mpeg1.1, mpeg1.2, and IL-10 genes in the brain, heart and liver of Danio rerio. Wherein, EVs secreted by THP-1 cells treated with TNF at doses of 10 and 20 ng/ml had opposite effects on the gene expression levels of il-1β in the brain, il-1β, il-10, and il-6 in the heart; on il-1β, il-10, mpx, and mpeg1.2 in the liver. EVs secreted by THP-1 cells treated with PMA at doses of 16 and 50 ng/ml had opposite effects on the expression levels of il-6 and il-10 genes in the heart and ifn-γ gene in the liver. EVs, produced by activated THP-1 cells have a systemic effect on Danio rerio manifested in a changing of the expression level of pro- and anti-inflammatory cytokine genes in the brain, liver, and heart. The qualitative composition of the EVs produced by activate THP-1 cells varies depending on the type and dose of the used stimulus, that reflects on strength and direction of the effects detected in vivo.

Effects of extended dietary supplementation with Santalum album essential oil on hemato-biochemical changes, innate immune response, antioxidant status, and expression of related gene in Nile tilapia (Oreochromis niloticus).

The effects of long-term dietary supplementation with sandalwood (Santalum album L.) essential oil (SEO) was investigated on hemato-biochemical biomarkers, immune status, antioxidant capacity, and resistance against Staphylococcus aureus in Nile tilapia (Oreochromis niloticus). Five groups (with four replicates) of O. niloticus (12.60 ± 0.20 g) were fed diets supplemented with SEO at doses of 0, 0.5, 1.0, 2.0, and 4.0 mL/kg diet for 60 days. Results indicated a substantial increase in blood protein levels and lower serum cholesterol, cortisol, glucose, urea, creatinine levels and, transaminase activities of fish fed a 2.0-mL SEO/kg diet. Serum lysozyme activity, nitric oxide, complement-3 levels, and phagocytic activity were significantly improved in O. niloticus after 60 days of feeding SEO-supplemented diets. Dietary SEO at level of 2.0-mL SEO/kg diet increased the activities of SOD, CAT, and GPx, and decreased MDA levels in liver homogenate. In addition, dietary 2.0-mL SEO/kg diet significantly upregulated antioxidant genes expression (CAT, SOD, GPx, GST, and GSR) with downregulation of apoptotic genes (HSP70, TLR2, caspase-3, and PCNA) in the liver. Furthermore, SEO-enriched diets significantly down-regulated pro-inflammatory (TNF-α, IL-1β, and IL-8) and up-regulated anti-inflammatory cytokine genes (TFG-β and IL-10) in the spleen. Moreover, SEO fortification increased the relative percentage of survival against S. aureus challenge and regulated immune-antioxidant genes in the spleen after the challenge. Overall, the results revealed that long-term using SEO might strengthen the physiological performance, hepatic oxidant/antioxidant balance, innate immune response, and resistance of O. niloticus against bacterial infections.

CHANGES IN THE EXPRESSION LEVEL OF PRO- AND ANTI-INFLAMMATORY CYTOKINE GENES IN THE BLOOD OF COWS DURING THE THERAPY OF CHRONIC ENDOMETRITIS

The article presents the material on studying the relative expression of pro- and anti-inflammatory cytokine genes in the blood of cows during the therapy of chronic endometritis in cows. It has been established that during treatment there is a decrease in the relative expression level of pro-inflammatory (TNF-α, IL-2) with a simultaneous increase in anti-inflammatory (IL-4, IL-10) cytokines. The greatest clinical efficacy of treatment of cows with chronic endometritis was established when using the antimicrobial drug "PK", biostimulgin and uteroton – 89.5%, accompanied by a decrease in the relative expression level of the TNFα genes in the blood by 2.1 times, IL-2 - by 2.18 times, with a simultaneous increase in IL-4 by 1.82 times, IL-10 – by 1.71 times. The dominance of the relative expression of anti-inflammatory cytokine genes over pro-inflammatory ones indicates the extinction of inflammatory processes in the uterus.

Chronic social defeat stress and glucocorticoid regulation in brain regions: resistance or hypersensitivity?

Chronic social defeat stress and glucocorticoid regulation in brain regions: resistance or hypersensitivity?

The ameliorative role of methionine in hepatic steatosis and stress response in juvenile black seabream (Acanthopagrus schlegelii) fed with a high-fat diet

Methionine (Met) is not only an essential amino acid of aquatic animals, but it also regulates lipid metabolism. To investigate the impacts of dietary Met supplementation on lipid metabolism, oxidative stress (OS), inflammation and apoptosis in marine fish, a feeding trial lasting for eight weeks was conducted on juvenile black seabream (Acanthopagrus schlegelii) (initial weight 1.61 ± 0.01 g) fed with five different experimental diets: Control, regular fat diet (11% lipid level); HFD, high-fat diet (17% lipid level), HFD contained different contents of Met (5, 10 or 20 g/ kg) named HFD + M1, HFD + M2, and HFD + M3, respectively. Serum biochemical indices results indicated that the contents of triglyceride (TG), total cholesterol (TC), non-esterified fatty acid (NEFA) and low-density lipoprotein cholesterol (LDL-C), as well as the activity of alanine transaminase (ALT), markedly decreased in HFD + M1 group compared to those in the HFD treatment. Results of liver sections indicated that lipid droplets dramatically decreased by dietary Met supplementation than the HFD group. Similar results were reflected in transmission electron microscope (TEM) images. The mRNA-abundant levels of adenosine monophosphate-activated protein kinase α (ampkα) and silent information regulator 1 (sirt1) significantly increased by dietary HFD + M1 than those in the HFD treatment. When comparing to the HFD treatment, the levels of gene expression for lipogenesis pathway genes, specifically sterol regulatory element-binding protein-1c (srebp-1c) and fatty acid synthase (fas) were markedly improved by dietary HFD + M1. In contrast, opposite results were recorded in the mRNA expression levels of lipolysis pathway related genes (pparα, cpt1a, hsl, atgl, lpl) that were down-regulated by the HFD + M1 by promoting related lipogenesis. The transcriptional expression level of srebp-1c was significantly suppressed in HFD + M1 diet. HFD caused oxidative stress, inflammation and apoptosis in the liver reflected in increasing the reactive oxygen species (ROS) and malonaldehyde (MDA) contents, c-Jun N-terminal kinase (jnk) and nuclear factor kappa B (nf-κb) mRNA expression levels, but those physiological stresses could be alleviated by dietary Met supplementation. Specifically, the antioxidant enzyme activities and gene expression levels were notably up-regulated by HFD + M1 group than those in fish fed with HFD. Additionally, compared to the HFD treatment, dietary Met supplementation significantly decreased the transcriptional expression levels of pro-inflammation and pro apoptosis genes, but up-regulated the transcriptional expression of anti-inflammatory cytokine and anti-apoptosis gene. Together, this study elucidates that HFD supplemented with Met can ameliorate hepatic steatosis, oxidative stress, inflammation and apoptosis induced by HFD, confirming that Met can play a vital role in lowing lipid accumulation by regulating lipid metabolism in A. schlegelii fed with HFD.

Contribution of polymorphic variants of pro- and anti-inflammatory cytokine genes to the development of occupational lung dust pathology in miners

Introduction. The formation and progression of occupational pathology of the bronchopulmonary system during prolonged contact with fibrogenic dust are closely related to changes in the concentration of certain pro- and anti-inflammatory cytokines in the bloodstream. The change in cytokine expression is due to the presence of polymorphism of certain genes, which results in quantitative or structural changes in proteins that can affect the course and outcome of dust lung diseases.&#x0D; The study aims to determine the associations of single-nucleotide polymorphisms of cytokine genes with the formation of occupational dust pathology of the lungs.&#x0D; Materials and methods. Scientists examined 140 coal mine workers who had been exposed to high concentrations of coal and stone dust for a long time. We divided the study participants into 2 groups. The main group included 75 miners with a previously established diagnosis of "dust pathology of the lungs", the comparison group consisted of 65 men working in the same sanitary and hygienic conditions, but without an established occupational disease of the bronchopulmonary system. The research groups are comparable by gender, age and ethnicity. The scientists carried out genotyping by allele-specific polymerase chain reaction in real time.&#x0D; Results. Experts have identified a number of gene polymorphisms associated with the risk of developing occupational pathology of the bronchopulmonary system as a result of exposure to fibrogenic dust in Kuzbass miners. They also found that the following markers predispose to the formation of occupational lung pathology in miners: allele C and heterozygous genotype C/T of the polymorphic locus rs1800896 of the IL-10 gene, allele C of the polymorphism rs1800795 of the IL-6 gene, homozygous genotype T/T of the IL-4 gene (rs2243250). In turn, the T allele and the homozygous T/T genotype of the rs1800896 polymorphism of the IL-10 gene, as well as the G allele of the rs1800795 polymorphic locus of the IL-6 gene have a protective effect on the development of occupational lung dust pathology.&#x0D; Limitations. The conducted study has limitations in the form of a relatively small sample size and the lack of quantitative determination of the concentration of cytokines in the blood.&#x0D; Conclusion. The results of the conducted studies indicate the contribution of polymorphic variants of the genes of the studied pro- and anti-inflammatory cytokines to the formation of genetic predisposition and resistance to the development of dust pathology of the lungs of professional genesis. With the help of the obtained informative markers, it is possible to assess the risk of developing dust lung diseases in miners working in contact with high concentrations of coal-rock dust.&#x0D; Ethics. The Bioethical Committee of the Research Institute of Complex Problems of Hygiene and Occupational Diseases has approved the study. The examination of patients was carried out on the basis of ethical standards developed in accordance with the Helsinki Declaration of the World Medical Association "Ethical principles of conducting Scientific medical research with human participation" as amended in 2013 and "Rules of Clinical Practice in the Russian Federation" approved by Order of the Ministry of Health of the Russian Federation No. 266 dated 06/19/2003. The study was conducted with the written consent of the subjects.

The Effects of Zinc Oxide Nanoparticles on Antioxidation, Inflammation, Tight Junction Integrity, and Apoptosis in Heat-Stressed Bovine Intestinal Epithelial Cells In Vitro.

Zinc oxide nanoparticles (nano-ZnO) have diverse applications in numerous biomedical processes. The present study explored the effects of these nanoparticles on antioxidation, inflammation, tight junction integrity, and apoptosis in heat-stressed bovine intestinal epithelial cells (BIECs). Primary BIECs that were isolated and cultured from calves either were subjected to heat stress alone (42°C for 6 h) or were simultaneously heat-stressed and treated with nano-ZnO (0.8 μg/mL). Cell viability, apoptosis, and expression of genes involved in antioxidation (Nrf2, HO-1, SOD1, and GCLM), inflammation-related genes (TLR4, NF-κB, TNF-α, IL-6, IL-8, and IL-10), intestinal barrier genes (Claudin, Occludin, and ZO-1), and apoptosis-related genes (Cyt-c, Caspase-3, and Caspase-9) were assessed to evaluate the effect of nano-ZnO on heat-stressed BIECs. The nanoparticles significantly increased cell viability and decreased the rate of apoptosis of BIECs induced by heat stress. In addition, nano-ZnO promoted the expression of antioxidant-related genes HO-1 and GCLM and anti-inflammatory cytokine gene IL-10, and inhibited the pro-inflammatory cytokine-related genes IL-6 and IL-8. The nanoparticles also enhanced expression of the Claudin and ZO-1 genes, and decreased expression of the apoptosis-related genes Cyt-c and Caspase-3. These results reveal that nano-ZnO improve the antioxidant and immune capacity of BIECs and mitigate apoptosis of intestinal epithelial cells induced by heat stress. Thus, nano-ZnO have potential for detrimental the adverse effects of heat stress in dairy cows.

Tea polyphenols protect bovine intestinal epithelial cells from the adverse effects of heat-stress in vitro

Heat-stress (HS) leads to impaired gut health, adversely affecting milk production of dairy cows. In the present study, we investigated the protective effects of tea polyphenols (TP) against HS-induced damage in bovine intestinal epithelial cells (BIECs) and explored the underlying mechanisms. Primary BIECs were isolated from bovine duodenum, cultured and treated as follows: (1) control cells incubated in complete medium at 37 °C for 12 h, (2) TP group incubated in medium containing 100 μg/mL TP at 37 °C for 12 h, (3) HS group incubated in medium at 37 °C for 6 h followed by 6 h at 42 °C, and (4) HS + TP group incubated with 100 μg/mL TP for 6 h at 37 °C and 6 h at 42 °C. TP improved cell viability and antioxidant capacity, and decreased apoptosis and LDH activity. TP led to upregulation of Nrf2 and its target antioxidant genes HO-1, NQO1 and SOD1 expression. TP significantly decreased the expression of proinflammatory cytokine genes (NF-κB, IL-6 and TNF-α), and increased expression of the anti-inflammatory cytokine gene, IL-10. The above results suggested that TP protected BIECs from HS-induced adverse effects by alleviating oxidative stress and inflammatory responses, indicating that TP can alleviate HS-induced intestinal damage in dairy cows.