Relative Expression Of TNF-α Research Articles

Correlation between albumin to fibrinogen ratio, C-reactive protein to albumin ratio and Th17 cells in patients with rheumatoid arthritis

BackgroundThe albumin to fibrinogen ratio (AFR) and the C-reactive protein to albumin ratio (CAR) have been served as inflammatory markers. However, their roles in RA remain unclear. We investigated the association of AFR/CAR with the concentration of autoantibodies and Th17 cells in RA. MethodsA total of 196 RA patients, 200 patients with systemic lupus erythematosus (SLE), and 200 healthy donors (HD) who were admitted to the First Affiliated Hospital of Fujian Medical University were enrolled. The results of FIB, ALB, CRP, anti-cyclic citrullinated peptide antibodies (anti-CCP), rheumatoid factor (RF) and erythrocyte sedimentation rate (ESR) from RA patients and SLE patients were retrospectively analyzed. The percentage of Th17 cells in peripheral blood of RA patients was detected by flow cytometry, and the relative expression of TNF-α, IL-6 and IL-17A was detected by RT-qPCR. Correlation analysis of AFR/CAR and Th17 cells, CRP, ESR, TNF-α, IL-6 and IL-17A in RA was conducted. ResultsCompared with SLE patients and healthy donors (HD), AFR concentration was significantly lower (P < 0.01) in RA patients, while CAR concentration was significantly increased (P < 0.01) in RA patients. AFR showed negative correlation with CRP (r = −0.7103), ESR (r = −0.6542), RF (−0.2219), Th17 cells (r = −0.5952) and IL-17A (r = −0.4681). CAR was positively correlated with CRP (r = 0.9899), ESR (r = 0.605), RF (0.1867), Th17 cells (r = 0.6818), TNF-α (r = 0.3388), and IL-17A (r = 0.2046). ConclusionsThe concentration of AFR in RA patients was reduced, while CAR concentration was increased. AFR and CAR are associated with CRP, ESR, RF, and Th17 cell ratios in RA patients, which can be used as potential indicators for determining RA inflammation.

Heat-killed Bacillus sp. SJ-10 probiotic acts as a growth and humoral innate immunity response enhancer in olive flounder (Paralichthys olivaceus)

Investigations were carried out to evaluate and quantify the effects of dietary supplementation with heat-killed (HK) Bacillus sp. SJ-10 (BSJ-10) probiotic (1 × 108 CFU g−1) on the growth and immunity of olive flounder (Paralichthys olivaceus). Flounder (averagely 9.64 g) were divided into two groups, and fed control and HK BSJ-10 (HKBSJ-10)-inoculated diets for 8 weeks. Investigations were carried out on growth and feed utilizations, innate immunity, serum biochemical parameters, microvilli length, and pro- and anti-inflammatory cytokine gene (tumor necrosis factor [TNF]-α, interleukin [IL]-1β, IL-6, and IL-10) transcriptions. Compared to control, HKBSJ-10 diet significantly (P < 0.05) enhanced weight gain and protein efficiency ratio, 1.17 and 1.11 folds respectively. Humoral innate immune parameters, lysozyme and superoxide dismutase in treatment group were also elevated by 1.34 and 1.16 folds. Similarly, an increased (P < 0.05) relative expressions of TNF-α, IL-1β, IL-6 were recorded in liver (2.71, 3.38, and 4.12 folds respectively), and gill (2.08, 1.98, and 1.81 folds respectively) than that of controls. Moreover, after challenge with Streptococcus iniae (1 × 108 CFU mL−1), the HKBSJ-10-fed group exhibited significantly higher protection (P < 0.05) against streptococcosis compared to controls, validating the observed changes in immune parameters and induction on the cytokine-encoding genes. Therefore, HKBSJ-10 increases growth, modulates innate immune parameters, and protects olive flounders against streptococcosis.

Effects of dietary oxidized fish oil on growth performance, antioxidant defense system, apoptosis and mitochondrial function of juvenile largemouth bass (Micropterus salmoides)

Four isonitrogenous and isolipidic diets containing fresh fish oil (peroxide value, POV: 7.2 meq kg−1, diet FR) and three degrees of oxidized fish oil (POV: 155, 275 and 564 meq kg−1, diet OX155, OX275 and OX564, respectively) were formulated to investigate the effects of dietary oxidized fish oil on growth performance, body composition, antioxidant defense system and mitochondrial function of juvenile largemouth bass, Micropterus salmoides. After a 12-week feeding trial, fish fed the OX155 obtained significantly higher (P < .05) weight gain, final body weight and specific growth rate than the FR because of their remarkable higher feed intake. Malonaldehyde (MDA) levels were markedly increased in plasma, liver and mitochondria of largemouth bass fed diet OX564 than those fed FR. Fish fed the OX564 also obtained the highest levels of lipid peroxidation (LPO) than other groups. Antioxidant enzyme activities and mRNA levels were stimulated after 12 weeks of feeding. The highest activities of superoxide dismutase (SOD) and glutathione peroxidase (Gpx) in liver and plasma were obtained in group OX564. The relative expression of TNF-α was decreased with the increasing oxidative degree of dietary fish oil, while the relative expression of Caspase 8, 9 and 10 were increased with the increasing oxidative degree of dietary fish oil. The relative expression of Bcl-2 and Bcl-xL were lower in fish fed the OX275 and OX564. The lowest activities of malate dehydrogenase (MDH), succinate dehydrogenase (SDH) and citrate synthase in liver were found in group OX564, in which the ATP level also lower than other groups. Liver mitochondria of fish fed the FR were more resistant to the calcium-induced mitochondrial swelling. More reactive oxygen species (ROS) were produced in fish fed the OX564 compared with those fed the FR after the 12-week trial. Membrane potential and Ca2+ induced swelling of mitochondria also affected by dietary oxidized fish oil. The present study indicated that dietary oxidized fish oil increased oxidative stress, influenced antioxidant system and induced apoptosis of largemouth bass, hepatic mitochondrial function and energy metabolism also were impaired by dietary oxidized fish oil.

Inhibitory effects of apolipoprotein M on inflammatory factors induced by high glucose in human retinal vascular endothelial cells

Objective To observe the expressing changes of apolipoprotein M (ApoM), tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) in human retinal vascular endothelial cells (HRECs) under the high glucose culture condition and investigate the inhibitory effects of ApoM overepression on the expressions of TNF-α and MCP-1. Methods HRECs were cultured in DMEM containing 10% fetal bovine serum and 5.5 mmol/L D-glucose and assigned to 6 groups.The cells in the normal control group were cultured in above culture medium; the cells in the high glucose group were treated using the DMEM with 30 mmol/L D-glucose; ApoM was transfected into the cells using lentiviral vector in the ApoM transfected group; lentiviral vector without ApoM sequence was transfected in the empty vector group; the cells transfected by empty vector were cultured in high glucose culture medium in the empty vector+ high glucose group; the cells in the ApoM transfection+ high glucose group were treated by ApoM sequence transfection and high glucose incubation.The relative expression of ApoM, TNF-α and MCP-1 mRNA was detected using real-time quantitative PCR, and the relative expression of ApoM protein was evaluated using Western blot assay. Results Compared with the normal control group, the mRNA expression levels of ApoM, TNF-α and MCP-1 in the high glucose group were significantly increased (t=5.517, 3.295, 2.555; all P<0.05). HRECs grew well after infected with lentivirus.The relative expression level of ApoM mRNA in the ApoM transfected group was 236.400±39.270, which was significantly higher than 1.000±0.153 in the empty vector group (t=5.995, P<0.01). An enhanced protein band of ApoM was seen in the ApoM transfected group, and the protein band was absent in the empty vector group.The relative expression band in the ApoM transfected group was 1.000±0.249 and 2.978±0.285 in the cells cultured with normal culture medium or high glucose culture medium, respectively, with a significant difference between them (t=5.056, P<0.01). The relative expressions of TNF-α and MCP-1 in the mRNA levels were significantly different among the empty vector group, empty vector+ high glucose group, ApoM transfected group and ApoM transfection+ high glucose group (F=5.966, P=0.026; F=14.410, P=0.002). Compared with the empty vector+ high glucose group, the relative expressions of TNF-α and MCP-1 mRNA were considerably reduced in the ApoM transfection+ high glucose group (P=0.017, 0.004). Conclusions High glucose environment up-regulates the expression of ApoM, MCP-1 and TNF-α in HRECs.Overexpression of ApoM inhibits the up-regulation of MCP-1 and TNF-α expression induced by high glucose. Key words: Apolipoprotein M; Glucose/pharmacology; Inflammation; Retina; Endothelial cells, vascular; Humans; Monocyte chemotactic factor-1; Tumor necrosis factor-α

Effects of Endogenous n-3PUFA on Body Weight, Autophagy and Inflammation in Mice

The fat-1 transgenic mice were used as models, to investigate the effects of n-3PUFAs on body weight, the expression of inflammation and autophagy in mice, and to explore its mechanism. The mice were divided into two groups: fat-1 transgenic mice and wild-type mice. The body weight and body length were measured and the index of body weight/body length was calculated regularly every week for 8 weeks. Cardiac blood was collected for determination of TG, CT, HDL-C, LDL-C and BG. Frozen sections of liver were stained by Oil Red O to observe the lipid droplets. The expression of autophagy proteins P62, LC3 and ATG7 in the hypothalamus were detected by western blot, and the relative quantitative analysis was performed. Real-time quantitative PCR was used to quantify the mRNA expression of TNF-α, IL-6, IL-1β, IFN-γ, MCP-1, TLR-4 and adiponectin in epididymal adipose tissue. The body weight/body length of fat-1 transgenic mice was significantly lower than that of wild-type mice(P<0.05), the levels of TG, CT, HDL-C, LDL-C and BG in the serum of fat-1 transgenic mice were significantly lower than those in wild-type mice(P<0.05). Lipid droplets in the liver of fat-1 transgenic mice were significantly less. The expression of P62 in fat-1 transgenic mice was significantly down-regulated (P <0.05), while the expression of ATG7 was significantly increased (P <0.05), and the ratio of LC3 Ⅱ / Ⅰ was significantly increased(P<0.05). The results of real-time quantitative PCR showed that the mRNA relative expression of TNF-α, IL-6, IL-1β, IFN-γ, MCP-1 and TLR-4 in epididymal fat tissue of fat-1 transgenic mice was significantly decreased, and the expression of adiponectin was increased (P < 0.05). n-3PUFAs reduce the body weight to prevent obesity may by up-regulation of hypothalamic autophagy, and down-regulation of inflammation in peripheral fat.

Red blood cell distribution width: a potential laboratory parameter for monitoring inflammation in rheumatoid arthritis.

Correlation analysis of red blood cell distribution width (RDW) and C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), tumor necrosis factor α (TNF-α), interleukin (IL)-6, and IL-10 in rheumatoid arthritis (RA) to investigate whether RDW can serve as a potential parameter for indicating inflammation in RA patients. A total of 670 RA patients from October 2014 to April 2016 were enrolled in our study. The white blood cell (WBC), red blood cell (RBC), platelet (PLT), hemoglobin (HGB), RDW, CRP, and ESR in peripheral blood of patients with RA were retrospectively analyzed. The relative expression of TNF-α, IL-6, and IL-10 was detected by RT-qPCR. Correlation analysis between RDW and CRP, ESR, TNF-α, IL-6, and IL-10 in RA was conducted by Microsoft Excel. RDW level was significantly increased in RA patients compared to osteoarthritis (OA) patients (P<0.001) and healthy donors (HDs) (P<0.001), and RDW was positively associated with inflammatory markers, such as CRP and ESR. In ROC curve analysis, the area under the curve (AUC) of RDW for the identification of RA was 0.881, with a 95% confidence interval (CI) from 0.864 to 0.898. Moreover, correlation analysis showed that RDW level was positively associated with TNF-α and IL-6, however negatively associated with IL-10. RDW was increased in patients with RA which was associated with inflammation in RA, suggesting that RDW may be a potential auxiliary marker for indicating inflammation process in RA conveniently.

Effect of four intestinal strains on TNF-α IL8 and IL6 expression in Caco-2 cells

Gut microbiota play an important roles in induction of various cellular responses, including cytokine production. In this work, we examined the involvement of four intestinal strains in induction of the cytokine response (the relative expression of TNF-α, IL-6, and IL-8) in intestinal epithelial cells. Caco-2 cells were treated with cell-free supernatants (CFS) of four intestinal strains: Shigella spp., Bacteroides fragilis, Weissella cibaria, Enterococcus faecium. Shigella spp., Bacteroides fragilis, Weissella cibaria, Enterococcus faecium CFSs resulted in marked an increase of mRNA TNF-α content ( p<0.05 or p<0.01).In additon, Shigella spp. CFS, Bacteroides fragilis CFS, Weissella cibaria CFS significantly increased expression mRNA for IL-8 ( p<0.05 or p<0.01), In contrast, Enterococcus faecium caused significant a decrease of mRNA IL-8(p<0.01). Last but not the least important, Shigella spp., Bacteroides fragilis, Weissella cibaria, Enterococcus faecium CFSs led to mark an increase of mRNA IL-6 level (p<0.01). These results indicate that four intestinal strains produce bioactive metabolites, suggesting that gut microbiota metabolites may therefore provide an entirely new approach towards protection of intestinal epithelial and treatment of human diseases.

Differential Expression of Th1- and Th2- Type Cytokines in Peripheral Blood Mononuclear Cells of Murrah Buffalo (Bubalus Bubalis) on TLR2 Induction by B. Subtilis Peptidoglycan

Peripheral blood mononuclear cells (PBMCs) discriminate microbial pathogens and induce T-cell responses of appropriate effector phenotype accordingly. Toll-like receptors (TLRs), in part, mediate this microbial recognition and differentiation while the development of T-cell effector functions critically depends on the release of Th1- or Th2- type cytokines. In the present study, buffalo PBMCs were stimulated under in vitro culture conditions by Bacillus subtilis cell wall petidoglycan, a TLR2 ligand, in a dose-and time- dependent manner. The expression of TLR2 as well as the subsequent differential induction of the Th1 and Th2 type cytokines was measured. Stimulation was analyzed across five doses of peptidoglycan (10 μ/ml, 20 μg/ml, 30 μg/ml, 40 μg/ml and 50 μg/ml) for 3 h, 12 h, 24 h and 36 h incubation periods. We observed the induction of TLR2 expression in a dose- and time-dependent manner and the peptidoglycan induced tolerance beyond 30 μg/ml dose at all incubation periods. The correlation between peptidoglycan stimulation and TLR2 induction was found positive at all doses and for all incubation periods. Increased production of all the cytokines was observed at low doses for 3 h incubation, but the expression of IL-4 was relatively higher than IL-12 at the higher antigen doses, indicating tailoring towards Th2 response. At 12 h incubation, there was a pronounced decrease in IL-4 and IL-10 expression relative to IL-12 in a dose- dependent manner, indicating skewing to Th1 polarization. The expression of IL-12 was highest for all doses across all the incubation intervals at 24 h incubation, indicating Th1 polarization. The relative expression of TNF-α and IFN-γ was also higher while that of IL-4 and IL-10 showed a decrease. For 36 h incubation, at low doses, relative increase in the expression of IL-4 and IL-10 was observed which decreased at higher doses, as did the expression of all other cytokines. The exhaustion of cytokine production at 36 h indicated that PBMCs became refractory to further stimulation. It can be concluded from this study that the cytokine response to sPGN initially was of Th2 type which skews, more pronouncedly, to Th1 type with time till the cells become refractory to further stimulation.

Banding or Burdizzo castration and carprofen administration on peripheral leukocyte inflammatory cytokine transcripts

The objective was to investigate if Banding or Burdizzo castration of bulls would alter the gene expression profile of a range of peripheral leukocyte inflammatory cytokines (IL-1, IL-6, IL-8, IL-10, interferon-γ and tumor necrosis factor-α) and to determine if the administration of carprofen (C) before castration would affect the expression of these genes. Thirty Holstein–Friesian bulls (5.5 months; Mean 191 ± (SEM) 3.7 kg) were blocked by weight and randomly assigned to one of five treatments: (1) untreated control (CON); (2) Banding castration at 0 min (BAND); (3) BAND following an i.v. injection of 1.4 mg/kg BW of carprofen (C) at −20 min (BAND + C); (4) Burdizzo castration at 0 min (BURD); or (5) BURD following 1.4 mg/kg BW of carprofen at −20 min (BURD + C). Blood samples were collected at 1 h before castration and 6, 24 and 48 h post-castration for routine hematology and quantitative real-time PCR analysis of cytokine gene expression analysis. Generally, there were no differences ( P > 0.05) among treatment groups in hematological variables following castration. Cortisol concentrations were unchanged throughout the experimental period in CON bulls. BURD animals had greater cortisol concentrations than BAND and CON animals at 6 h post treatment. Transitory effects were observed only in the expression of IL-6 and TNF-α. The relative expression of IL-6 was greater in the BURD than in the BAND treatment ( P < 0.05) at 24 h post-castration and was greater in the BURD + C group than in the BURD group ( P < 0.05) at 48 h. The relative expression of TNF-α was greater in BAND than in the BURD group ( P < 0.05) at 48 h. In conclusion, these findings indicate that Banding or Burdizzo castration did not have any major effect on peripheral leukocyte inflammatory cytokine gene expression; Banding castration caused a greater pro-inflammatory cytokine gene expression reaction than Burdizzo castration and carprofen administration can affect IL-6 gene expression levels in BURD castrated animals.