Clinical Implications of Tumor Necrosis Factor-Alpha, Interleukin-6 and Resistin in Coronary Artery Disease

Tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6) are involved in the progression of coronary artery disease (CAD). The cytokines’ levels are associated with the severity of CAD. We have recently reported on the association of resistin, a relatively novel cytokine with the pathogenesis of cardiovascular disease (CVD). Although the inflammatory cytokines’ impact on atherosclerosis is widely accepted, yet some controversy exists regarding the involvement of these factors in atherogenesis. The current review highlights the potential association of TNF-alpha, IL-6 and resistin SNPs (single nucleotide polymorphisms) with CAD. Molecular genetics data along with the intracellular signaling cascade mechanisms may have important clinical implications in the treatment of CAD.

Tumor Necrosis Factor-Alpha (TNF)-308G/A and Interleukin 8(IL-8)-251C/T Polymorphisms in Pulmonary Tuberculosis Patients from Congo

Background: Tuberculosis (TB) is one of the world’s deadliest infectious diseases. Tumor necrosis factor-Alpha (TNF-α) and Interleukin 8 (IL-8) are involved in the pathogenesis of pulmonary TB (PTB). However, the contribution of polymorphisms of these cytokines to PTB susceptibility needed more investigation across geographic regions and ethnic groups. Purpose: The aim of this study was to investigate the association of the TNF-α-308 G/A and IL-8-251T/A polymorphisms with PTB risk in the Congolese population. Methods: This case-control study included 150 PTB patients and 160 control subjects. Blood samples were collected from all participants and were used for the TNF-α-308 G/A and IL-8-251T/A genotyping by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. Odds ratios (OR) were calculated to estimate the potential polymorphism associations. A P level of Results: A significant difference was found between PTB patients and controls regarding the TNF-α-308AA genotype (P = 0.035) distribution. Moreover, this genotype was associated with risk to TB (OR = 7.19, 95% CI = 0.85 - 60.65, P = 0.035). The A allele was significantly more frequent in PTB patients than in controls, and was associated with risk to PTB (OR = 1.68, 95% CI = 1.05 - 2.68, P = 0.014). Regarding the IL-8-251T/A gene, TA and AA genotypes were significantly more frequent in PTB patients compared to controls, and were associated with increased risk to PTB (OR = 2.64, 95% CI = 0.97 - 7.18, P = 0.031 and OR = 3.0, 95% CI = 1.13 - 7.98, P = 0.014, respectively). However, the IL-8-251 A allele was not associated to PTB susceptibility (OR = 0.27, 95% CI = 0.15 - 0.44). Conclusion: TNF-α-308G/A and IL-8-251T/A polymorphisms may be associated to PTB susceptibility in the Congolese population, and the AA genotype of both cytokines could be a risk factor.

Plasma levels of tumor necrotic factor-alpha and interleukin-6, -8 during orthotopic liver transplantation and their relations to postoperative pulmonary complications

BACKGROUND: Pulmonary complications after orthoto- pic liver transplantation (OLT) include high morbidity and mortality. Experimental data have suggested hepatic ische- mia and reperfusion are induced by pro-inflammatory cyto- kines. The high level of inflammatory cytokines might ad- ditionally influence pulmonary cappillary fluid filtration. The objectives of this study were to measure the concentra- tions of tumor necrotic factor-alpha (TNF-α), interleukin- 6 (IL-6) and interleukin-8 (IL-8) during OLT and to in- vestigate the relationship between these cytokines and post- operative pulmonary complications. METHODS: Twenty-two patients undergoing OLT were divided into two groups according to whether they had postoperative pulmonary complications: group A consis- ting of 8 patients with postoperative pulmonary complica- tions , and group B consisting of 14 patients without post- operative pulmonary complications. Enzyme-linked im- munoassay (ELISA) was used to determine serum TNF-α, IL-6 and IL-8. Blood samples were taken at the beginning of operation (T0 ), clamping and cross-clamping of the in- ferior cava and portal vein (T1, T2 ), 90 minutes and 3 hours after reperfusion (T3 , T4 ) and 24 hours after opera- tion (T5). RESULTS: The level of PaO2/FiO2 in group A was lower than that in group B ( P <0. 05 ). The concentrations of TNF-α, IL-6 and IL-8 in the two groups increased rapidly at T2 , peaked at T3 , decreased rapidly after T3 until 24 hours after operation. The concentrations of TNF-α, IL-6 and IL-8 in group A were higher than those in group B at T2, T3, and T4(P<0.05). CONCLUSION: After un-clamping of the inferior cava and portal vein, the serum concentrations of TNF-α, IL-6 and IL-8 increased may be related to pulmonary injury after he- patic ischemic reperfusion.

Serum Concentrations of Angiotensin, C-Reactive Protein, Interleukin-8, and Tumor Necrosis Factor-α in Train Driver Population

Train drivers are engaged in high-stress job. It may induce sleep, fatigue, and alertness loss at work, and endanger public safety. It’s unclear that cytokines of train driver would be influenced by their job. The research considers the hypothesis that stressful professions, such as train driver, influence the body’s immune system through the long-time and high-pressure working, and change production of neuro-immune factors. Using enzyme linked immunosorbent assay (ELISA), several neuro-immune factors were assayed among train drivers (N = 82) and health blood donors (N = 80) enrolled in the Yunnan Collaborative Innovation Center for Public Health and Disease Control. The concentrations of angiotensin, C-reactive protein (CRP), interleukin-8 (IL-8), and tumor necrosis factor-alpha (TNF-α) were determined. Kruskal-Wallis test and Dunn’s multiple comparisons test were performed for overall comparison between groups and for pairwise comparison, respectively. Statistical significance level was set at P < 0.05. The profession of train driving was not associated with significant increases or decreases in the systemic levels of inflammatory (CRP, IL-8, and TNF-α), but it was associated with the high expression of angiotensin in vivo. These findings suggest that the job of train driving may not be associated with significant alterations in systemic immune condition, but arouse the level of angiotensin.

Changes of TNF_α IL-6 and IL-8 in plasma and subeschar edema fluid of guinea pigs after thermal injury

To investigate the levels of cytokines in subeschar edema fluid (SEF). Methods: A guinea pig model with III°30% total body surface area (TBSA) was designed. SEF was collected serially at the time of escharectomy for up to 72 h after scald injury. The levels of tumor necrosis factor.(TNFα), interleukin-6 (IL-6) and interleukin-8 (IL-8) were determined by using the enzyme-linked immunosorbent assay (ELISA). Results: The levels of TNFα,IL-6 and IL-8 in plasma and SEF increased in the early phase of scald injury. Furthermore, the contents of TNFα， IL-6 and IL-8 in SEF were higher than those in plasma.Conclusion: The findings indicate that the subeschar edema fluid is a biologically active reservoir, and is an important participant in the postburn pathophysiologic process.

Relationship between serum levels of IL-4,IL-8,TNF-alpha,T cell subsets and prognosis in patients with psoriasis vulgaris

Objective:To study the relationship between serum levels of IL-4,IL-8,TNF-alpha,T cell subsets and prognosis in patients with psoriasis vulgaris.Methods:A total of 120 patients with psoriasis vulgaris who were treated in our hospital from January 2018 to January 2019 were selected as the study group,and 50 normal subjects who underwent health examination in our hospital during the same period were selected as the control group.The levels of serum IL-4,IL-8,TNF-alpha and T cell subsets in the observation group and the control group were detected and compared.The levels of IL-4,IL-8,TNF-alpha and T cell subsets in the observation group at different time after treatment were compared after standardized western medicine treatment.Pearson test was used to analyze the correlation between IL-4,IL-8,TNF-alpha and T cell subsets.Results:The levels of IL-4,IL-8 and TNF-a in the observation group were higher than those in the control group,while the levels of CD3+,CD4+,CD4+/CD8+in the observation group were lower than those in the control group,and the levels of CD8+in the observation group were higher than those in the control group.There was a significant difference between the two groups.In the observation group,after 8 weeks of treatment,the levels of IL-4,IL-8 and TNF-alpha continued to decrease,CD3+,CD4+,CD4+/CD8+increased and CD8+decreased with the prolongation of treatment time.There was significant difference among the groups.Pearson correlation test was used.IL-4,IL-8,TNF-a had negative correlation with CD3+,CD4+,CD4+/CD8+,and positive correlation with CD8+.Conclusion:The incidence of psoriasis vulgaris is related to the elevation of IL-4,IL-8,TNF-alpha levels and immunodeficiency.The prognosis of psoriasis vulgaris can be judged by monitoring the levels of IL-4,IL-8,TNF-alpha and T cell subsets.

Effects of Remifentanil Pretreatment on Inflammatory Response in Rats with Acute Cerebral Ischemia Injury

[Objectives]This study was conducted to investigate the effects of remifentanil pretreatment on inflammatory factors in rats with acute cerebral ischemia.[Methods]Sixty SD rats were randomly divided into the normal control group,sham operation group,ischemic brain injury group,and remifentanil pretreatment group.Except the normal control group,each group was divided into three subgroups(six in each group)according to the sampling time points of 6,12 and 24 h after execution.After modeling,the rats were scored for neurological deficit,and observed for pathological changes of neurons in the brain tissue by HE staining and the brain infarct volume by TTC staining,and the expression levels of TNF-α,IL-6 and IL-8 were detected by RT-PCR.[Results]HE staining:No significant changes were observed in the pathological morphology of the brain tissue in the blank group and sham operation group;and the neuronal structure of rats in the acute cerebral ischemia group was obviously damaged,and the neuronal damage in the remifentanil pretreatment group was less than that in the acute cerebral ischemia group at each time point.TTC staining:The gray brain infarct area in the remifentanil pretreatment group was significantly smaller than that in the cerebral ischemia group(P<0.05).RT-PCR detection results:The expression levels of TNF-α,IL-6 and IL-8 in the blank group and sham surgery group did not show significant changes at different times(P>0.05);and compared with the cerebral ischemia group,the expression levels of TNF-α,IL-6,and IL-8 in the remifentanil pretreatment group were significantly reduced at all time points(P<0.05).[Conclusions]Remifentanil pretreatment could protect the brain by reducing the expression of inflammatory factors after cerebral ischemia injury.

Sperm lipid peroxidation and pro-inflammatory cytokines

Aim： To investigate if interleukin-6 （IL-6）, interleukin-8 （IL-8）, interleukin- 10 （IL-10）, interferon-gamma （IFN-γ） or tumor necrosis factor-alpha （TNF-α） are able to stimulate the level of lipid peroxidation of sperm membranes, either alone or in the presence of leukocytes. Methods： Semen samples from normozoospermic donors were prepared by density gradient. The sperms were exposed to the indicated cytokines, at physiological and infection-inflammation concentrations, in the absence or presence of leukocytes. Lipid peroxidation of the sperm membranes was determined by measuring malondialdehyde （MDA） and 4-hydroxialkenals （HAE） formation. Results： TNF-α, IL-8 and IFN-γ increased the level of sperm membrane lipid peroxidation when tested at physiological concentrations. At infectioninflammation concentrations, only IL-8 was able to produce a higher effect. When assayed in the presence of leucocytes, IL-8 and TNF-α showed a higher effect at infection-inflammation concentrations than at physiological concentrations. Finally, IL-8 showed a higher effect in the presence of leukocytes than in their absence at both physiological and infection-inflammation concentrations. TNF-α also showed a higher effect when assayed in the presence of leuko- cytes than in their absence, but only at infection-inflammation concentrations. There was no effect of IL-6 or IL-10 in any of the tested conditions. Conclusion： Several pro-inflammatory cytokines at physiological concentrations increase the level of lipid peroxidation of sperm membranes, which could be important for the sperm fecundation process. However, infection-inflammation concentrations of some cytokines, such as IL-8 and TNF-α, either alone or in the presence of leukocytes, could drive the lipid peroxidation of the spermatozoa plasma membrane to levels that can affect the sperm fertility capacity.

EXPERIMENTAL WORK and RESEARCH Effect of Reduqing(热毒清) on TNF-α,IL-1,IL-6,IL-8 and PAF Levels in Endotoxin-Induced DIC Model of Rabbits

Objective: To study the mechanism of anti-endotoxemic effect of Reduqing Injection (RDQ)and to explore the essence of traditional Chinese 'heat-clearing and detoxifying therapy'. Methods: A disseminated intravascular coagulation (DIC) model was made in rabbits by intravenous injection with E. colt endotoxin. Increased plasma levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), interleukin-8 (IL-8) and platelet activating factor (PAF), as well as the ex-vivo LPS-induced interleukin-1 (IL-1 ) production in peripheral blood monocytes were observed in model (M) group. Results: Same parameters were significantly lower in RDQ and dexamethasone (DXM) groups than those in the M group (P < 0. 01, P < 0. 05). Histopathological examination showed severe damage of the liver, lungs and kidneys in the rabbits of M group, in contrast, only mild affects were seen in the RDQ and DXM groups. Conclusions: RDQ exhibits protective effect on rabbits against endotoxin-induced DIC. The suppression of cytokines and inflammatory mediator PAF by RDQ may play a central role in the inhibition of endotoxin-induced DIC cascade.

Changes of Cytokines in Serum and Whey of Dairy Cows with Subclinical Mastitis

In order to study the correlation of different degrees of subclinical mastitis and cytokines,the changes of interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-8(IL-8)and tumor necrosis factor-α(TNF-α)in blood and milk and whether the changes of these indexes were caused by mastitis were analyzed.The 40 Holstein cows in lactation period were tested and grouped by somatic cell count(SCC)method in milk:Group C(SCC<5×10~5 cells·mL^(-1)),Group I(5×10~5 cells·mL^(-1)5×10~6 cells·mL^(-1)).The contents of IL-1β,IL-6,IL-8 and TNF-αwere analyzed by radioimmunoassay.The results showed that the contents of IL-6,IL-8 and TNF-αin the blood and milk of different degrees of subclinical mastitis increased with the severity of mastitis,and were higher than those in normal group.The contents of IL-6,IL-8 and TNF-αin serum and whey of dairy cows affected by different degrees of subclinical mastitis were higher than those in normal group(Group C).The content of IL-1βincreased and decreased with the increase of inflammation,and it did not have the value of evaluating mastitis.In conclusion,the severities of cow subclinical mastitis and the contents of cytokines were positive correlations.These indexes could be used as the basis for judging mammary gland injury.

Effects of the 24 N-terminal Amino Acids of p55PIK on Endotoxinstimulated Release of Inflammatory Cytokines by HaCaT Cells

Summary： This study aimed to examine the effect of the 24 N-terminal amino acids （N24） ofp55PIK, a regulatory subunit of phosphatidylinositol 3-kinase （PI3K）, on the endotoxin lipopolysaccharide （LPS）-stimulated release of the cytokines （CKs） by HaCaT cells. The fusion protein, trans-acting activator of transcription （TAT）-N24 （an experimental peptide, EP） containing the N24 of PI3K-p55PIK, was constructed, and TAT-N24 fusion peptide was expressed and identified in BL21 E.coli. HaCaT cells （a human keratinocyte cell line） was cultured and stimulated by LPS at 100 ng/mL for 1, 2, 4, 8, 16 or 24 h, or by LPS at 10, 100 ng/mL, 1, 10 or 100 μg/mL of for 4 h. Changes in the protein and mRNA levels of tumor necrosis factor-alpha （TNF-ct）, interleukin-6 （IL-6） and interleukin-8 （IL-8） released by HaCaT cells following EP intervention were determined by enzyme-linked immunosorbent assay （ELISA） and real-time polymerase chain reaction （PCR）. Immunofluorescence confocal laser scanning microscopy was utilized to detect the protein expression and translocation of the p65 subunit of nuclear factor kappa-light-chain-enhancer of activated B cells （NF-r,B p65） in HaCaT cells. The expression of the NF-kB inhibitor alpha （Iv, B-a） protein in LPS-stimulated HaCaT cells after the EP intervention was measured by Western blotting. The resillts showed that EP treatment increased TNF-a secretion from HaCaT cells. EP at certain concentrations could effectively inhibit the LPS-stimulated release of TNF-a, IL-6 and IL-8 from HaCaT cells. The ELISA assay demonstrated that the concentrations of TNF-a, IL-6 and IL-8 in the supernatants of LPS-stimulated cells were reduced from 208.06±30.18, 86.4±9.78 and 260.59±54.05 pg/mL to 121.78±22.26, 53.18±7.36 and 125.08±35.17 pg/mL, respectively, in the supernatants of cells treated by LPS and EP combined. Real-time PCR also revealed that the expression of the three pro-inflammatory CKs was significantly decreased after EP intervention. Immunofluorescence confocal laser scanning microscopy showed that NF-kB p65 protein was primarily expressed in the cytoplasm of non-stimulated HaCaT cells. After LPS stimulation, NF-kB p65 was translocated into the nucleus, and the nuclear expression of this protein increased. The nuclear NF-kB p65 protein expression was inhibited after the addition of EP. Western blotting showed that Ir, B-a expression began to decrease 30 min after LPS stimulation and declined to a trough 4 h later. Ir, B-a expression began to gradually recover 16 h after LPS stimulation but remained at a lower-than-normal level at 24 h. Greater Ir, B-a expression was found in cells treated with LPS and EP combined than those treated with LPS alone. It was concluded that EP can effectively inhibit the LPS-stimulated expression of TNF-a, IL-6, and IL-8, which involves the inhibition of the hydrolysis of Ir, B-a and thereby blockage of the nuclear transloca- tion of NF-kB p65.

Effects of different dialytic membranes and dialysates on cytokine productions in patients undergoing hemodialysis

This study was to investigate the effects of different dialytic membranes and dialysates on cytokine production in patients undergoing hemodialysis and their relationship with hernodialytic complications. Two dialytic membranes (cuprophane and polysulfone membranes ) and two dialysates (bicarbonate and acetate ) were used for hemodialysis. Twenty-one patients were divided into 3 groups and dialyzed respectively with cuprophane acetate (CU-A), cuprophane bicarbonate (CU-B), and polysulfone bicarbonate (PS-B). It was found that the incidence of complications was the highest and lowest in itU A group and PS-B group respectively. The levels of plasma IL-6,IL-8 and TNFα were significantly increased in CU-A and CU-B groups in the lst h (P<0. 01), further increased in the 4th h (P<0. 001 ) and then decreased to the pre-dialytic levels in the 24th h after hemodialysis. These parameters were significantly higher in CU-A group than in CU-B group (P<0. 05). No significant changes of these cytokines were found in PS-B during and after hemodialysis. The plasma level of acid glycoprotein (AG) was significantly increased in CU-A and CU B groups in the 4th h after hemodialysis (P<0.05) and markedly correlated to the plasma levels of cytokines. α-AP level was remarkably higher in CU-A group than in CU-B group (P<0. 05)but there were no significant changes in PS-B group. our findings suggest that systemic release of such cytokines as IL-6, IL-8 and TNFα in patients undergoing hemodialysis might be related to the bio compatibility of dialytic membranes and dialysates and responsible for the side effects of hemodialysis. Polysulfone membrane and bicarbonate are relatively better materials for hemodlalysis. Improvement of bio-compatibility of the dialytic membranes and dialysates is helpful to reduce hemodialytic complications.

The Effect of Chloroquine on Pro-Inflammatory Cytokines Levels in Graves’ Disease: Historical Cohort from a Pilot Randomized Controlled Trial

Objectives: Analyzing the trend in the serum inflammatory cytokines levels in a historical cohort of patients treated with combination of chloroquine and methimazole. Material and methods: We analyzed the pro-inflammatory serum cytokines level [Interleukin-6(IL-6), Tumor Necrosis Factor alpha (TNF-α), Interleukin 1 alpha (IL-1 α) and Interferon gamma (INF-γ)] in the stored blood samples of 22 patients with Graves’ disease who previously randomized to receive either chloroquine and methimazole combination therapy or methimazole monotherapy. Total T3, T4 and TSH levels were measured by an enzyme linked immunosorbent assay (ELISA) method (DRG, New York, USA) and the result was published previously. In this study we used an ELISA method (Bender Medsystem Vienna Austria) to measure serum pro-inflammatory cytokines in the first 6 months of trial. Results: No significant differences in serum cytokines concentration were observed between the two treatment groups (p > 0.05). Although it was not statistically significant, serum INF-gamma concentration tended to be lower in the chloroquine group after four months of therapy (p = 0.12). Conclusion: In this study we found changes in the serum thyroid hormones level did not accompany concomitant changes in the serum cytokines levels in two treatment groups. Therefore it is possible that chloroquine reduce serum thyroid hormones levels independent of its immunomodulatory effect.

Effect of Reduqing (热毒清) on Endotoxin-Induced Production of Tissue Factor and Cytokines in Human Whole Blood

Objective: TO study the effect of Reduqing (RDQ) Injection on the lipopolysaccharide (LPS)induced tissue factor and cytokine production in whole blood. Methods: Heparinized human blood was incubated with LPS in the presence or absence of RDQ. The plasma concentrations of TNF-α, IL-1β, IL-6, and IL-8were measured by enzyme-linked immunosorbent assays (ELISA) and the monocyte tissue factor activity wasmeasured by a one-stage tissue factor induced plasma clotting time assay. Results: RDQ was found to diminishthe LPS-induced increase of TNF-α, IL-1β and IL-6 in plasma but did not completely abolish their production.In contrast to the effect on these cytokines, RDQ caused further increase of the plasma level of IL-8 and themonocyte TF activity in the presence of LPS. Conclusions: In the in vztro whole blood assay system used inthis study, the decrease of LPS-induced production of TNF-α, IL-1β, and IL-6 was similar to a previous in vivostudy on the effect of RDQ on the production of these cytokines in response to two-time LPS injection in rabbits, while the increase of IL-8 and TF production was contradictory to the previous in vivo study. Potentialreasons contributing to the differences are discussed.