Gene Expression Levels Of IL-1β Research Articles

Suppressive Effects of β-Hydroxybutyrate Administration on Lipopolysaccharide-Induced Inflammation in Broiler Chickens.

This study aimed to evaluate the suppressive effects of β-hydroxybutyrate (BHB) administration on lipopolysaccharide (LPS)-induced inflammation in broiler chickens. Twenty-day-old male broiler chickens were randomly allocated to three groups, each of which was treated with saline (control), intraperitoneal administration of LPS [1.5 mg/kg body weight (BW), Escherichia coli O127:B8], or LPS plus BHB (3 mmol/kg BW). Plasma albumin and total protein concentration were significantly reduced by LPS administration, while BHB co-treatment partially attenuated the effects. The LPS treatment significantly induced plasma aspartate and alanine aminotransferase activities, and interleukin (IL)-6 concentration, with the increases suppressed by BHB co-treatment (p < 0.05). The LPS treatment significantly increased the gene expression levels of IL-1β, IL-6, and IL-18 in the spleen and peripheral blood monocytes (PBMC), while the increases were partially attenuated by BHB in the spleen. Relatively higher levels of BHB dehydrogenase 1 and succinyl-CoA:3-ketoacid CoA transferase were observed in the spleen and skeletal muscle, while these gene levels were lower in PBMC and the liver. The present results suggest that BHB can suppress LPS-induced inflammation, in which ketolytic enzyme expression levels may be involved in broiler chickens.

NLRP3 inflammasome pathway involved in the pathogenesis of metabolic associated fatty liver disease

The prevalence of Metabolic-associated fatty liver disease (MAFLD) has been steadily increasing worldwide, paralleling the global epidemic of obesity and diabetes. It is estimated that approximately one-quarter of the global population is affected by MAFLD. Despite its high prevalence, MAFLD often goes undiagnosed due to the lack of specific symptoms in its early stages. However, as the disease progresses, it can lead to more severe liver-related complications such as fibrosis, cirrhosis, and hepatocellular carcinoma. Therefore, we aimed to investigate the expression levels of the nucleotide-binding oligomerization domain, leucine-rich repeat (LRR)—containing proteins (NLR) family pyrin domain-containing protein 3 [NLRP3] inflammasome pathway components, NLRP3 and interleukin 1β (IL-1β) genes in patients with MAFLD with various degrees of steatosis and fibrosis. Participants were classified into two equal groups; MAFLD group: consisted of 120 patients with different degrees of hepatic fibrosis and steatosis based on fibro scan results. The non-MAFLD group was comprised of 107 participants. Molecular analysis of pyrin domain-containing protein 3 and IL-1β relative gene expressions was performed in the blood of all participants, using Real-time quantitative polymerase chain reaction (RT-qPCR). Patients with post-MAFLD hepatic fibrosis had significantly higher relative gene expression levels of IL-1β and NLRP3; with IL-1β > 1.1 had AUC of 0.919, sensitivity of 88.33, specificity of 96.26, PPV of 96.4, and NPV of 88 and 92.3 accuracy (p value < 0.001). NLRP3 > 1.33 had a sensitivity of 97.5, specificity of 99.07, PPV of 99.2, NPV of 97.2, and 98.3 accuracy with an AUC of 0.991 (p value < 0.001) as predictors of post-MAFLD hepatic fibrosis.. A significant increase in the mean relative gene expression levels of both IL-1β and NLRP3 found in patients with early fibrosis (F0-F1-2); 31.97 ± 11.8 and 6.76 ± 2.18, respectively; compared with patients with advanced hepatic fibrosis stages (F2-F3); 2.62 ± 3.71 and 4.27 ± 2.99 (p < 0.001 each). The present study provides novel evidence for the possible involvement of IL-1β and NLRP3 inflammasome in metabolic-associated fatty liver disease pathogenesis and could be valid markers for the early detection of post-MAFLD hepatic fibrosis.

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.

Exopolysaccharide secreted by Lactiplantibacillus plantarum Y12 showed inhibitory effect on the pathogenicity of Shigella flexneri in vitro and in vivo

Shigella flexneri is a prevalent foodborne and waterborne pathogen that threatens human health. Our previous research indicated that the Lactiplantibacillus plantarum Y12 exopolysaccharide (L-EPS) potentially inhibited the pathogenicity of S. flexneri. The in vitro results of this study demonstrated that L-EPS effectively mitigated the symptoms induced by S. flexneri in HT-29 cells, including inhibited gene expression levels of IL-1β, IL-6, IL-8, TNF-α, TLR 2/4, and NOD1/2; decreased apoptosis ratio; and alleviated damage degree of intestinal barrier function (Zona occludens 1, Occludin, and Claudin-1). The in vivo results demonstrated that S. flexneri treated with L-EPS elicited mild adverse physiological manifestations, an inflammatory response, and tissue damage. The infection of S. flexneri caused significant alterations in the abundance of phylum (Firmicutes, Bacteroidota, Actinobacteriota, and Proteobacteria), family (Lachnospiraceae, Muribaculaceae, Rikenellaceae, Prevotellaceaea, Ruminococcaceae, and Lactobaillaceae), and genus (Escherichia Shigella and Lachnospirillaceae NK4A136 group) within the cecal microbiota. These changes were accompanied by perturbations in taurine and hypotaurine metabolism, tricarboxylic acid (TCA) cycle activity, arginine biosynthesis, and histidine metabolic pathways. However, intervention with L-EPS attenuated the dysbiosis of cecal microbiota and metabolic disturbances. In summary, our research suggested a potential application of L-EPS as a functional food additive for mitigating S. flexneri infection.

УРОВЕНЬ ЭКСПРЕССИИ ГЕНОВ ИММУННОГО СТАТУСА В КРОВИ У ПОРОСЯТ-ГИПОТРОФИКОВ

The article presents data on the level of expression of immune defense genes in normotrophic and hypotrophic piglets. 20 sows of 3–4 farrows and the offspring obtained from them were selected for the experiment. The piglets underwent clinical examination and weighing. Not reaching 800 g animals were counted as piglets-hypotrophic, animals over 800 g – respectively normotrophic. In the blood of normotrophic and hypotrophic piglets, the gene expression levels of IL-1β, TGF-β and TNF-α have minor changes: an increase in hypotrophic piglets by 1.2 and 1.1 times. However, the level of expression of the IFN-α gene in hypotrophic piglets was 4.2 times higher than in normotrophic piglets, which may be due to the immune response against the background of malnutrition, namely possible pyrogenicity.

Analysis of histopathology and changes of major cytokines in the lesions caused by Mycoplasma ovipneumoniae infection

BackgroundMycoplasma ovipneumoniae (M. ovipneumoniae) is one of the main pathogens of sheep pneumonia, causing a series of clinical symptoms, such as depression, anorexia, hyperthermia, cough, dyspnea, and tract secretions. In recent years, the prevalence of M. ovipneumoniae pneumonia has become increasingly serious in sheep farms in Ningxia, China, leading to the death of sheep, and causing significant economic losses. In this study, the pathological organs infected by M. ovipneumoniae were collected to observe histopathological change, to determine the tissue localization of M. ovipneumoniae, and to analyze the cytokine changes, which lays a basis for the diagnosis and pathogenesis of M. ovipneumoniae disease.ResultsIn this study, M. ovipneumoniae was detected in 97 of 105 samples collected from 13 large-scale sheep farms for nucleic acid by PCR. One representative isolate per farm was isolated from 13 farms. The lesions caused by M. ovipneumoniae were mainly in the trachea, bronchus, and lung, including necrosis of tracheal mucosal epithelial cells, disintegration of some epithelial cells, edema of mucosal lamina propria, with inflammatory cell infiltration, cytoplasmic vacuolization of epithelial cells of bronchial mucosa, massive infiltration of inflammatory cells in the alveolar space of lung, necrosis and hyperplasia of alveolar epithelial cells. Immunohistochemical analysis showed that the proportion of M. ovipneumoniae positive area in the lung was the largest, followed by that in the bronchus and trachea. Compared to healthy animals, diseased animals exhibited up-regulated gene expression levels of IL-1β, IL-6, and NF-κB in the trachea, bronchus, and lungs. In contrast, the expression of IL-10, IL-12, and IFN-γ was primarily limited to the trachea and bronchus. The expression of IL-1β showed differential patterns across different lung regions, with variations observed among lung lobes. Additionally, other cytokines consistently showed significant up-regulation specifically in the bronchus.ConclusionsM. ovipneumoniae is primarily found in the lungs of infected individuals. NF-κB, an essential transcription factor, is involved in the regulation of IL-1β transcription. IL-12 may enhance the cytotoxic function of natural killer cells during M. ovipneumoniae infection. Those findings demonstrate the distinct expression profiles of cytokines in various anatomical sites throughout disease progression, suggesting the potential role of bronchial tissue as a major site of immune response.

Effects of fermented soybean meal substitution for fish meal on intestinal flora and intestinal health in pearl gentian grouper.

This study explored the role of replacing fish meal protein with fermented soybean meal (FSBM) protein on the growth performance and intestinal morphology, immunity, and microbiota of the pearl gentian grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂). Three isonitrogenous and isolipidic diets with increasing levels of FSBM (0%, 20% and 40%; referred to as FM, FSBM20 and FSBM40 diets, respectively) as a replacement for fish meal were selected for this study. The pearl gentian grouper were fed these diets for 10 weeks. The findings revealed that the growth of fish fed the FSBM diets (FSBM20 and FSBM40) were remarkably lower than the fish fed the FM diet. Pathological manifestations of intestinal inflammation, such as shortened intestinal mucosal folds and thickened lamina propria, were observed in the fish fed the FSBM diets. Moreover, the gene expression levels of IL1β, IL12, IL17, and TNFα were remarkably upregulated in fish fed the FSBM40 diet, in contrast to the gene expression levels of IL4, IL5, IL10, and TGFβ1, which were remarkably downregulated (p < 0.05). The FSBM diets significantly affected the stability of the fish gut microbiota. Photobacterium was the dominant phylum in all experimental groups, and the proportion of these bacteria gradually decreased with increasing FSBM substitution. The composition of intestinal flora at the genus level was not the same in the three experimental groups, with a richer composition of intestinal bacteria detected in the FSBM20 and FSBM40 groups (p < 0.05). The correlation between intestinal flora balance and immune gene expression revealed that only Photobacterium was negatively correlated with the above upregulated genes, while other bacteria were positively correlated with these pro-inflammatory factors (p < 0.05). Photobacterium was positively correlated with the above downregulated genes, while other bacteria were negatively correlated with these anti-inflammatory factors (p < 0.05). In conclusion, high levels of substitution of FSBM for fish meal causes intestinal inflammation in pearl gentian grouper. This is likely associated with changes to the intestinal flora. More attention should be paid to the negative role of dietary FSBM on intestinal flora.

The Role of NLRP3 Inflammasome in Obesity and PCOS-A Systematic Review and Meta-Analysis.

Inflammasomes have recently been implicated in the pathogenesis of several chronic inflammatory disorders, such as diabetes and obesity. The aim of this meta-analysis was to investigate the possible role of the NLRP3 inflammasome in obesity and polycystic ovarian syndrome (PCOS). A comprehensive search of electronic databases was conducted to identify studies investigating NLRP3 its related components (Caspase 1, ASC and IL-1β) in adipose tissue and/or blood from obese individuals compared to non-obese controls. Another search was conducted for studies investigating NLRP3 in PCOS women and animal models. The ssearched databases included Medline, EMBASE, Cochrane Library, PubMed, Clinicaltrials.gov, the EU Clinical Trials Register and the WHO International Clinical Trials Register. The quality and risk of bias for the included articles were assessed using the modified Newcastle-Ottawa scale. Data were extracted and pooled using RevMan software for the calculation of the standardized mean difference (SMD) and 95% confidence interval (CI). Twelve eligible studies were included in the obesity systematic review and nine in the PCOS review. Of the obesity studies, nine (n = 270) were included in the meta-analysis, which showed a significantly higher adipose tissue NLRP3 gene expression in obese (n = 186) versus non-obese (n = 84) participants (SMD 1.07; 95% CI, 0.27, 1.87). Pooled analysis of adipose tissue IL-1β data from four studies showed significantly higher IL-1β gene expression levels in adipose tissue from 88 obese participants versus 39 non-obese controls (SMD 0.56; 95% CI, 0.13, 0.99). Meta-analysis of adipose tissue ASC data from four studies showed a significantly higher level in obese (n = 109) versus non-obese (n = 42) individuals (SMD 0.91, 95% CI, 0.30, 1.52). Of the nine PCOS articles, three were human (n = 185) and six were animal studies utilizing PCOS rat/mouse models. All studies apart from one article consistently showed upregulated NLRP3 and its components in PCOS women and animal models. In conclusion, obesity and PCOS seem to be associated with upregulated expression of NLRP3 inflammasome components. Further research is required to validate these findings and to elucidate the role of NLRP3 in obesity and PCOS.

EVALUATION OF RESISTANCE AND GENE EXPRESSION OF BARRAMUNDI, Lates calcarifer POST-INFECTION OF NERVOUS NECROSIS VIRUS

The most common problem in barramundi Lates calcarifer seedling production is the high mortality (&gt; 90%) caused by nervous necrosis virus (NNV) infection. This research aims to evaluate the resistance and gene expression of barramundi challenged by NNV. Two populations were used in this study, i.e., Australian, and Situbondo-originated barramundi populations. The immune-related gene expression levels in the liver, head of kidney, and spleen were observed at 48 and 96 hours after post-infection (hpi). Barramundi’s survival and blood parameters were evaluated post-NNV infection. The results showed that the highest survival was revealed in Situbondo’s barramundi (42.0±4.47%) compared to Australian barramundi (20.0±7.07%) and no mortality was observed in the control without NNV infection. The higher survival rate in barramundi from Situbondo was in line with the blood profile. The number of red blood cell from Situbondo barramundi post-NNV infection (ST) at 96 hpi was higher (P&lt;0.05) than Australian barramundi post-NNV infection (AT). The number of white blood cell of ST at 48 hpi was higher (P&lt;0.05) than AT, but started to decrease at 96 hpi in ST barramundi. The total white blood cell in AT barramundi increased from 48 to 192 hpi. TNFα and IL1-β gene expression levels were significantly higher in the liver, head kidney, and spleen of Situbondo compared to Australian barramundi at 48 hpi, while MHCIIα gene expression in Situbondo’s was significantly higher compared to Australian barramundi at 96 hpi. These results indicate the important roles of all the genes in the barramundi’s immune responses against viral infection. Based on the results of the research, Situbondo’s barramundi has the potential to be used as a candidate for generating broodstock of disease-resistant strain.

Neutralizing peripheral circulating IL1β slows the progression of ALS in a lentivirus-infected OPTNE478G mouse model.

Amyotrophic lateral sclerosis (ALS) is irreversible and fatal within 3-5 years, with limited options for treatment. It is imperative to develop a symptom-based treatment that may increase the survival of ALS patients and improve their quality of life. Inflammation status, especially elevated interleukin 1β (IL1β), has been reported to play a critical role in ALS progression. Our study determined that neutralizing circulating IL1β slows down the progression of ALS in an ALS mouse model. The ALS mouse model was developed by microinjection of lentivirus-carrying OPTNE478G (optineurin, a mutation from ALS patients) into the intra-motor cortex of mice. Peripheral circulating IL1β was neutralized by injecting anti-IL1β antibody into the tail vein. Enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR) were carried out to determine the protein and gene expression levels of IL1β. TUNEL assay was used to assess the neural cell death. Immunofluorescent staining of MAP2 and CASP3 was accomplished to evaluate neuronal cell apoptosis. Glial fibrillary acidic protein staining was performed to analyze the number of astrocytes. Rotarod test, grip strength test, balance beam test, and footprint test were conducted to assess the locomotive function after anti-IL1β treatment. The model revealed that neuroinflammation contributes to ALS progression. ALS mice exhibited elevated neuroinflammation and IL1β secretion. After anti-IL1β treatment, ALS mice revealed decreased neural cell death and astrogliosis and gained improved muscle strength and motor ability. Blocking IL1β is a promising strategy to slow down the progression of ALS.

A study of the potential effect of dietary fishmeal replacement with cricket meal (Gryllus bimaculatus) on growth performance, blood health, liver antioxidant activities, intestinal microbiota and immune-related gene expression of juvenile channel catfish

Studies have shown that insect meals can partially or completely replace fish meal (FM) that are commonly used in aquaculture. Therefore, a 10-week feeding trail was done to test the potential impacts of replacement of FM with cricket meal (CM) on the growth performance, body composition, blood health, liver antioxidant activities, intestinal microbiota and immune reaction of juvenile channel catfish. Five isonitrogenous (crude protein 35%) and isocaloric (19 MJ/kg−1) diets were formulated to have 0%, 25%, 50%,75% and 100% CM and named T0, T25, T50, T75 and T100 groups, respectively. Final body weight (FBW), weight gain rate (WGR) and specific growth rate (SGR) in fish fed T50 and T75 groups were considerably elevated compared with fish fed control T0 group. Higher hepatosomatic index (HSI) was found in fish fed T50, T75 and T100 groups compared with fish fed control T0 group. The moisture and crude protein contents in fish fed T75 and T100 groups were significantly increased than that of fish fed control T0 group. Hematological parameters examination of juvenile channel catfish demonstrated that hemoglobin (HGB), hematocrit (HCT), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) showed significantly higher values with increasing content of CM in dietary fish fed groups. Malondialdehyde (MDA) content and glutathione peroxidase (GPx) activity level were highest in fish fed T75 group, respectively. Catalase (CAT) activity in fish fed T50 to T100 groups was significantly greater than those of fish fed control T0 and fish fed T25 groups. Intestinal microbiota sequencing analysis showed that Chao1, Shannon, Simpson indices, and Good’s coverage in fish fed control T0, fish fed T50 and T100 groups were not significantly affected. Intestinal microbiota analysis showed the relative abundance of Proteobacteria and Firmicutes were dominated at the phyla level. Moreover, with an increasing of CM replacement, the abundance of a prominent pathogenic bacterial Acinetobacter was reduced at the genus level although no significant differences. The immune-related gene expression levels of IL-1β, IL-8, IL-10 and HIF1α were up-regulated in intestine when CM replacement increased from 0% to 75%, and higher levels were observed in fish fed T75 group comparing to others. The expression levels of TNF-α, IL-22 and IFN-γ were significantly elevated in fish fed T100 group. Notably, fish fed T50 to T100 groups down-regulated the expression of NF-κB compared to the other groups. To conclude, the present findings show that CM could enhance growth performance, improving blood health and liver antioxidant capacity, and substantial alterations in intestinal microbial composition of juvenile channel catfish, suggesting that FM could be replaced by CM as insect protein source in channel catfish diet.

Selenomethionine protected BMECs from inflammatory injury and oxidative damage induced by Klebsiella pneumoniae by inhibiting the NF-κB and activating the Nrf2 signaling pathway

Klebsiella pneumoniae (K. pneumoniae) is one of the main environmental pathogens causing bovine mastitis. The incidence of bovine mastitis caused by K. pneumoniae is increasing worldwide. Selenium is an essential trace element that has multiple physiological functions, such as antioxidant and anti-inflammatory activities. Therefore, this study aimed to verify whether selenomethionine (SeMet) could contribute to alleviating the inflammatory injury and oxidative damage induced by K. pneumoniae. Bovine mammary epithelial cells were cultured in vitro and pretreated with 4 μM SeMet before being infected with K. pneumoniae. Western blot analysis was used to detect the expression of the related proteins in the NF-κB and Nrf2 signaling pathways. The gene expression levels of IL-1β, IL-6, IL-8, TNF-α, Nrf2, Keap1, NQO-1 and HO-1 were detected using RT–qPCR. The levels of MDA, GSH-PX, SOD, CAT and T-AOC were detected by commercial assay kits. Flow cytometry was used to determine the level of intracellular ROS, and immunofluorescence was used to detect the nuclear localization of Nrf2 protein. Briefly, SeMet downregulated the phosphorylation levels of IκBα and p65 proteins and the gene expression levels of IL-1β, IL-6, IL-8 and TNF-α were also decreased. Moreover, the protein and gene expression levels of Nrf2, NQO-1 and HO-1 were upregulated, and the nuclear expression of Nrf2 protein was also promoted, which enhanced the activity of antioxidant enzymes. In conclusion, SeMet protected BMECs from inflammatory injury and oxidative stress induced by K. pneumoniae by inhibiting the NF-κB and activating the Nrf2 signaling pathway.

Effect of dietary lipid levels on the anti-oxidant responses, initial immunity, and mTOR signaling in the liver of coho salmon (Oncorhynchus kisutch)

In the present study, we evaluated the effects of dietary lipid levels on the anti-oxidant responses, initial immunity, and mTOR signaling in the liver of coho salmon Oncorhynchus kisutch. The post-larval coho salmon were fed four isonitrogenous experimental diets with crude lipid levels of 6.6%, 10.56%, 18.91%, and 26.91% for 12 weeks, respectively. Compared to the control group (6.6% dietary lipid level), the activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), peroxidase (POD), glutathione peroxidase (GSH-PX), and total antioxidant capacity (TAC) were enhanced by 10.56% and 18.91% dietary lipid levels in the liver of coho salmon. However, no significant difference was observed on these anti-oxidant enzyme activities when the fish were fed 26.91% lipid level diet. Compared to the control group (6.6% dietary lipid level), the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA) as well as the generation of hydroxy radical (·OH) and superoxide anion (O2·-) were decreased by 10.56% and 18.91% dietary lipid levels. In addition, the gene and protein expression level of PI3K, Akt, and mTOR was induced by 10.56%, 18.91%, and 26.91% dietary lipid levels. The gene expression level of IL-1β, IL-6, and TNFα was also increased with the increase of dietary lipid levels. The result of hepatic tissue sections showed that the cellular morphology of liver was affected by the higher dietary lipid levels. In conclusion, the dietary lipid levels affected the anti-oxidant responses, initial immunity, and mTOR signaling in the liver of post-larval coho salmon.

BN82002 alleviated tissue damage of septic mice by reducing inflammatory response through inhibiting AKT2/NF-κB signaling pathway

BN82002 is well-known as an inhibitor of the CDC25 phosphatase. However, it was recently reported that BN82002 also selectively suppressed AKT2 and reduced inflammatory responses in lipopolysaccharide (LPS)-stimulated macrophage-like RAW264.7 cells. Therefore, in this study, we evaluated the alleviating efficacy of BN82002 in sepsis in vivo. BN82002 (50 μM) suppressed the mRNA levels of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 in LPS-treated peritoneal macrophages without cytotoxicity. The septic in vivo mouse model was established on the basis of the endotoxin model using poly(I:C) (10 mg/kg) and LPS (54 mg/kg). In histological analysis, peritoneal injection of BN82002 (20 mg/kg) significantly reduced lung, kidney, and liver damage. Lung edema and serum alanine transaminase (ALT), aspartate transaminase (AST), TNF-α, IL-1β, and nitric oxide (NO) levels also were decreased by BN82002 (20 mg/kg). In addition, BN82002 (20 mg/kg) suppressed the mRNA levels of TNF-α in lung and liver tissues. Gene expression levels of IL-1β and IL-6 were decreased in lung, kidney, and liver in the BN82002 (20 mg/kg) group. Furthermore, p-AKT2 and p-IκBα levels were reduced by BN82002 (20 mg/kg). Finally, all septic mice died 7 days after poly(I:C)/LPS-injection, whereas 4 mice in the BN82002 (20 mg/kg) group, survived strongly suggesting that BN82002 reduces sepsis mortality. In conclusion, we verified that pre-treatment with BN82002 protects against tissue damage and increases survival by inhibiting AKT2-NF-κB signaling in septic mice. These results suggest that BN82002 could be utilized in the treatment of sepsis.

Effects of dietary antimicrobial peptides on intestinal morphology, antioxidant status, immune responses, microbiota and pathogen disease resistance in grass carp Ctenopharyngodon idellus

This study aims to highlight the effects (8 weeks) of dietary antimicrobial peptides (AMPs, a compound of 6 kDa and 5 kDa from intestine) on intestinal morphological functions and health status in grass carp (Ctenopharyngodon idellus). Fish were supplemented with various gradient concentrations of AMPs, including M0 (0 mg/kg), M1 (100 mg/kg), M2 (200 mg/kg), M3 (400 mg/kg), M4 (800 mg/kg) and M5 (1600 mg/kg). Our results showed that amylase, lipase, chymotrypsin enzymatic levels, and total antioxidant capacity (T-AOC) were significantly increased (p < 0.05), while malondialdehyde (MDA) content was significantly decreased in the intestines of the AMP treated groups compared to the M0. Histological analysis revealed villus height and crypt depth of foregut and midgut in the M4 group were significantly different (p < 0.05) compared to the M0. In the M3 group, the gene expression levels of IL-1β were significantly up-regulated, while levels of IL10 and TGF-β were significantly down-regulated than other treated and control groups. The abundance of Firmicutes was significantly increased (p < 0.05), while the Planctomycetes abundance was decreased at phylum level in M1-M5 groups. Subsequent to the AMP treatment, fish were injected with Aeromonas. hydrophila to assess disease resistant potential. In A. hydrophila injected M3-group, the gene expressions of IL-1β, IL8, and TNF-α were significantly down-regulated while that of TGF-β was significantly up-regulated, and IL10 showed no significant difference compared to the control. Further, AMPs also increased the abundance of the Acidobacteria, Proteobacteria, and Patescibacteria, and decreased the abundance of the Fusobacteria and Firmicutes. Therefore, dietary AMPs (400–800 mg/kg) boosted intestinal health by promoting intestinal morphology, digestive and antioxidant capacities, immunity, and intestinal microbiota in C. idellus.

Firmicutes/Bacteroidetes Ratio in the Gut Microbiota and IL-1β, IL-6, IL-8, TLR2, TLR4, TLR5 Gene Expressions in Type 2 Diabetes.

The aim of this study was to determine the Firmicutes/Bacteroidetes ratio in the gut microbiota and IL-1β, IL-6, IL-8, TLR2, TLR4 and TLR5 gene expression levels in the blood of adult type 2 diabetes (T2D) patients and compare it with that of adult nondiabetic healthy controls (HC). Between May 2016 and April 2017, 99 T2D patients and 99 HCs were enrolled in the study. Bacteroidetes and Firmicutes levels were assessed from stool sample DNA and IL-1β, IL-6, IL-8, TLR2, TLR4, and TLR5 gene expression levels assesed from blood sample RNA via qPCR from both T2D patients and healthy controls. The Firmicutes/Bacteroidetes ratio detected in the stool of type 2 diabetes patients was found to be higher with a statistically significant difference (p < 0.0001). Gene expression levels of IL-1β, IL-6, IL-8, TLR2, TLR4, and TLR5 were found to be upregulated. The highest upregulation was detected in IL-6 with 11 fold in T2D patients comparing with HCs. F/B ratio and gene expression levels were elevated in T2D patients. Firmicutes were positively correlated with studied gene expressions. A better understanding of the complex interaction between gut microbiota, environment, and diabetes will allow for more effective prevention and treatment strategies for T2D.

Anti-Inflammatory, Antioxidant, and Antifibrotic Effects of Gingival-Derived MSCs on Bleomycin-Induced Pulmonary Fibrosis in Mice.

Background: Mesenchymal stem cell (MSC) intervention has been associated with lung protection. We attempted to determine whether mouse gingival-derived mesenchymal stem cells (GMSCs) could protect against bleomycin-induced pulmonary fibrosis. Methods: Mice were divided into three groups: control (Con), bleomycin (Bl), and bleomycin + MSCs (Bl + MSCs). Mice were treated with 5 mg/kg bleomycin via transtracheal instillation to induce pulmonary fibrosis. We assessed the following parameters: histopathological severity of injury in the lung, liver, kidney, and aortic tissues; the degree of pulmonary fibrosis; pulmonary inflammation; pulmonary oedema; profibrotic factor levels in bronchoalveolar lavage fluid (BALF) and lung tissue; oxidative stress-related indicators and apoptotic index in lung tissue; and gene expression levels of IL-1β, IL-8, TNF-α, lysophosphatidic acid (LPA), lysophosphatidic acid receptor 1 (LPA1), TGF-β, matrix metalloproteinase 9 (MMP-9), neutrophil elastase (NE), MPO, and IL-10 in lung tissue. Results: GMSC intervention attenuated bleomycin-induced pulmonary fibrosis, pulmonary inflammation, pulmonary oedema, and apoptosis. Bleomycin instillation notably increased expression levels of the IL-1β, IL-8, TNF-α, LPA, LPA1, TGF-β, MMP-9, NE, and MPO genes and attenuated expression levels of the IL-10 gene in lung tissue, and these effects were reversed by GMSC intervention. Bleomycin instillation notably upregulated MDA and MPO levels and downregulated GSH and SOD levels in lung tissue, and these effects were reversed by GMSC intervention. GMSC intervention prevented upregulation of neutrophil content in the lung, liver, and kidney tissues and the apoptotic index in lung tissue. Conclusions: GMSC intervention exhibits anti-inflammatory and antioxidant capacities. Deleterious accumulation of neutrophils, which is reduced by GMSC intervention, is a key component of bleomycin-induced pulmonary fibrosis. GMSC intervention impairs bleomycin-induced NE, MMP-9, LPA, APL1, and TGF-β release.

Influence of Intracanal Cryotherapy on Inflammatory Mediators Expression using Different Irrigation Protocols: A Randomized Clinical Trial

ABSTRACTAim: To evaluate the effect of intra-canal cryotherapy on interleukin-1βand interleukin-10 expression in teeth with symptomatic apical periodontitis.Methodology: Forty-eight single rooted teeth in patients with symptomatic apical periodontitiswere randomly divided into 4 groups according to the irrigation protocol.Group I (control); 5% sodium hypochlorite at room temperature. Group II; 5% coldsodium hypochlorite (2-5°C). Group III; final rinse with 20 mL of 5% cold sodiumhypochlorite (2-5°C) for 5 minutes. Group IV; final rinse with 20 mL of cold saline (2-5°C) for 5 minutes. Paper point passing 2 mm beyond the apex was used to collect periapicalfluid before, immediately after mechanical instrumentation and after one week tocharacterize the mRNA expression of IL-1β and Il-10 using real-time polymerase chainreaction (PCR). Statistical analysis was performed using one-way ANOVA to comparebetween all groups. Results: Our results revealed statistically significant (at P≤ 0.05)down regulation of IL-1β gene expression levels in all groups after one week with nostatistically significant difference between the four groups and statistically significant(at P≤ 0.05) up regulation of IL-10 gene expression levels in all groups after one week.Conclusion: All the cryotherapy irrigation protocols showed lower levels of expressionof the pro-inflammatory mediators (interleukin-1β) and higher levels of expression ofthe anti-inflammatory mediator (interleukin-10) than the control group after one week.

Ex Vivo Perfusion With Methylprednisolone Attenuates Brain Death-induced Lung Injury in Rats

Background.The onset of brain death (BD) leads to the deterioration of potential donor lungs. Methylprednisolone is considered to increase lung oxygenation capacity and enhance the procurement yield of donor lungs, when applied in situ, during donor management. However, whether BD-induced lung damage is ameliorated upon treatment with methylprednisolone during acellular ex vivo lung perfusion (EVLP), remains unknown. We aimed to investigate whether the quality of lungs from brain-dead donors improves upon methylprednisolone treatment during EVLP.Methods.Rat lungs were randomly assigned to 1 of 3 experimental groups (n = 8/group): (1) healthy, directly procured lungs subjected to EVLP; (2) lungs from brain-dead rats subjected to cold storage and EVLP; and (3) lungs from brain-dead rats subjected to cold storage and EVLP with 40 mg methylprednisolone added to the perfusate. Ventilation and perfusion parameters, histology, edema formation, metabolic profile, and inflammatory status of lungs were investigated.Results.Methylprednisolone treated lungs from brain-dead donors improved positive inspiratory pressures needed to maintain tidal volumes of 7 mL/kg of body weight, which was 25.6 ± 5.8 cm H2O in untreated lungs and 18.0 ± 3.0 cm H2O in methylprednisolone treated lungs, after 6 h EVLP. Furthermore, dynamic lung compliance increased upon methylprednisolone treatment, with values of 0.11 ± 0.05 mL/cm H2O versus 0.18 ± 0.04 mL/cm H2O after 6 h of EVLP. Methylprednisolone treatment ameliorated the amount of lung edema, as corroborated by a reduction of 0.7 in the wet/dry ratio. Although glucose consumption levels were comparable, the BD-induced cumulative lactate production decreased from 0.44 ± 0.26 to 0.11 ± 0.16 mmol/L upon methylprednisolone treatment. Finally, BD-induced inflammatory status was reduced upon methylprednisolone treatment compared to untreated lungs from brain-dead donors, as reflected by lower proinflammatory gene expression levels of IL-1β, IL-6 and MCP-1, and IL-6 perfusate levels.Conclusions.We showed that methylprednisolone treatment during EVLP attenuates BD-induced lung injury.

A novel dietary multi-strain yeast fraction modulates intestinal toll-like-receptor signalling and mucosal responses of rainbow trout (Oncorhynchus mykiss).

This study was conducted to evaluate the mucosal immune responses of rainbow trout when supplementing an experimental formulated feed with multi-strain yeast fraction product (Saccharomyces cerevisiae and Cyberlindnera jardinii). In total, 360 fish (initial BW 23.1 ± 0.2 g) were randomly allotted into three dietary treatments in an 8-week feeding trial. The dietary treatments included basal diet (control) and control + 1.5 g/kg multi-strain yeast fraction product (MsYF) fed continuously and pulsed every two weeks between control and MsYF diet. No negative effects on growth performance of feeding the MsYF supplemented diet were observed. SGR and FCR averaged 2.30 ± 0.03%/day and 1.03 ± 0.03, respectively, across experimental groups. Muscularis thickness in the anterior intestine after 8 weeks of feeding was significantly elevated by 44.3% in fish fed the MsYF continuously, and by 14.4% in fish fed the MsYF pulsed (P < 0.02). Significant elevations in goblet cell density in the anterior and posterior (>50% increase) intestine were observed after 8 weeks of feeding the MsYF supplemented diet (P< 0.03). In contrast, lamina propria width was significantly lower in fish fed the experimental diets (>10% reduction). The gene expression analysis of the intestine revealed significant elevations in expression of tlr2, il1r1, irak4, and tollip2 after 4 weeks of feeding the MsYF. Significant elevations in effector cytokines tnfα, il10 and tgfβ were observed after 4 weeks of feeding the MsYF regime. After 8 weeks significant elevations in the gene expression levels of il1β, ifnγ, and il12 were observed in fish fed the MsYF. Likewise, the expression of the transcription factor gata3 was significantly elevated (P<0.01). Supplementation of the multi-strain yeast fraction product positively modulates the intestinal mucosal response of rainbow trout through interaction with toll-like receptor two signalling pathway and potential for increased capacity of delivery of antigens to the underlying mucosal associated lymphoid tissue.