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10% of labeled tumor cells) of TNF receptor 1 (TNFR1), the protein product of TNFRSF1A gene, was correlated with sarcomatoid dedifferentiation and was an independent predictive factor of clinically unfavorable response and shorter survivals in separated TKI-treated ccRCC cohort.CONCLUSION: TNF-α signaling may play a role in TKI resistance, and TNFR1 expression may serve as a predictive biomarker for clinically unfavorable TKI responses in ccRCC.
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Humans , Biomarkers , Carcinoma, Renal Cell , Cohort Studies , Dataset , Drug Resistance , Gene Expression , Gene Expression Profiling , Heterografts , Immunohistochemistry , Protein-Tyrosine Kinases , Receptors, Tumor Necrosis Factor , Receptors, Tumor Necrosis Factor, Type I , Tumor Necrosis Factor-alphaABSTRACT
Objective::To explore the therapeutic effect and mechanism of Chaige Qinlian Tang on pneumonia in young mice. Method::The pneumonia model was duplicated by slowly dripping Staphylococcus aureus into the nasal cavity of mice.After successful modeling, the mice were randomly divided into model group, clindamycin group, and high and low-dose Chaige Qinlian Tang groups, with sham operation group as negative control group.The rats were given 200 mg·kg-1 high-dose Chaige Qinlian Tang, 100 mg·kg-1 low-dose Chaige Qinlian Tang and 120 mg·kg-1 clindamycin.The mice were observed every day.Colonies were counted in the lungs of each group five days later.The expression levels of interleukin(IL)-16, tumor necrosis factor (TNF)-α in lung lavage fluid of each group were determined by enzyme linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) and Western blot were used to measure the expression levels of IL-16, TNF-α in lung lavage fluid of each group.The expressions of tumor necrosis factor receptor (TNFR) 1, Caspase-3 and Caspase-7 in lung and the pathological changes of lung were observed. Result::Compared with the sham operation group, the respiratory state and the activity state of the model mice were worse, and the survival rate was higher in the high-dose Chaige Qinlian Tang group.Compared with the sham operation group, the pulmonary colony counts in the model group and treatment groups were increased, compared with the model group, the lung colony counts in clindamycin group and high-dose Chaige Qinlian Tang group were improved significantly (P<0.05, P<0.01). Compared with the control group, the expression levels of IL-16, TNF-α, TNFR1, Caspase-3, Caspase-7 mRNA and protein in the lung of model group and treatment groups were significantly increased (P<0.01). Compared with model group, the expression levels of IL-16, TNF-α and TNFR1, Caspase-3, Caspase-7 in the lung of clindamycin group and high and low-dose Chaige Qinlian Tang groups were significantly increased (P<0.01). The expression levels of protein and mRNA were significantly decreased (P<0.05, P<0.01), and the pathological changes of lung were improved, especially in clindamycin group and high-dose Chaige Qinlian Tang group. Conclusion::Chaige Qinlian Tang has a certain therapeutic effect on Staphylococcus aureus pneumonia in young mice.This effect may be related to regulating TNFR1, Caspase-3 and Caspase-7 pathways, reducing the secretion of IL-16 and TNF-alpha, and enhancing the clearance of staphylococcus aureus.
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Objective: To study the effect of small molecule compounds of Hedysari Radix in ntagonizing tumor necrosis factor receptor type 1 (TNFR1) based on molecular docking. Method: The structure of small molecular compound of Hedysari Radix was downloaded from the chemical composition compound library of traditional Chinese medicine, and then optimized to obtain the composition compound library of Hedysari Radix. The three-dimensional structure of the inflammatory target TNFR1 (PDB ID:1TNR) was identified. After hydrotreating and anhydrating, the binding pocket residues were identified according to the literature. According to the defined target structure and binding pocket, the flexible molecular docking was conducted between the composition compound library and the target, and the score (Glide Score) was obtained. Based on the results of molecular docking, the first nine small molecular compounds of Glide Score were selected as candidate components. On this basis, the drug-likeness was analyzed, which involved small molecular compounds that meet the number of hydrogen-bonded receptors, the number of hydrogen-bonded donors, the formula weight, the number of rotatable key and the numerical range of lipo-hydro partition coefficient. Finally, the binding mode was analyzed according to pharmacokinetic parameters and complex structure of composition-target docking. Result: The residue set in the TNFR1 drug-binding pocket were identified as glutamic acid109 (Glu109), lysine 35(Lys35), alanine62 (Ala62), serine 74 (Ser74), lysine75 (Lys75), cysteine76 (Cys76), argnine 77(Arg77), glutamine82 (Gln82), threonine89 (Thr89), asparticacid91 (Asp91), argnine92 (Arg92), aspartic acid93 (Asp93), threonine 94(Thr94), valine95 (Val95), cysteine 96(Cys96), argnine104 (Arg104), tyrosine106 (Tyr106), asparagine110 (Asn110), leucine111 (Leu111), phenylalanine112(Phe112), glutamic acid 131(Glu131) and lysine132 (Lys132). Totally 43 small molecular compounds of Hedysari Radix were obtained. Five small molecular compounds, namely hedysari radix, quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin, were screened by comprehensive factors, like docking scoring. Conclusion: Quercetin, isoliquiritin, naringenin, calycosin and liquiritigenin are the effective anti-inflammatory substances of Hedysari Radix, with a great possibility of becoming TNFR1 antagonists.
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Objective To explore effects of ginsenosides Rg1 on the expression of poly(ADP-ribose) polymerase-1 (PARP-1) and tumor necrosis factor receptor (TNFR) 1 in cortex cells after focal cerebral ischemia in rats. Methods Ninety healthy rats were randomly divided into sham-operative group, focal cerebral ischemia group, ginsenoside Rg 1groups (low, medium and high concentrations) and drug control group. Rats were intraperitoneally injected saline 45 mg/kg, saline 45 mg/kg+ginsenosides Rg1 10, 20 and 40 mg/kg, nimodipine 1 mg/kg 5 d before surgery, respectively. Focal cerebral isch?emia model was made by middle cerebral artery occluding in rats. The neurological deficit score and TTC staining were used to verify the success of the rat model. The expressions of PARP-1 and TNFR1 were evaluated by immunohistochemical meth?od and Western blot technique. Results There were obvious symptoms of neurological deficit and large pale infarct area in focal cerebral ischemia group compared with those of sham-operative group. There were higher percentages of neurological deficit score and infarct area in ginsenosides Rg1 groups and positive control group than those of sham-operative group, but which were lower than those of ischemia group (P<0.05). There were no significant differences between ginsenosides Rg1 groups and positive control group. The positive cells of PARP-1 and TNFR1 were higher in ginsenosides Rg1 low-dose group than those of sham-operative group and positive control group, while ones of medium and high-dose Rg1 group were higher than those of sham-operative group, and were lower than those of ischemia group (P<0.05). Compared with sham-op?erative group, PARP-1 and TNFR1 expression strips were significantly enhanced in ischemia group. Expression strips were higher in ginsenosides Rg1 low-dose group than those of sham-operative group. Expression strips were higher in ginsen?osides Rg1 medium-dose group than those of sham-operative group, but which were lower than those of ischemia group, and ones of high-dose group were lower than ischemia group (P<0.05). Conclusion Ginsenoside Rg1 shows protective effects on focal ischemia injury, which may be related with down-regulation of the expression of PARP-1 and TNFR1.
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Previous study on TNFRl-mediated hepatocyte apoptosis has been implicated in the development of fulminant viral hepatitis.To interfere with the potentially effective target,plasmid named p-mTNFR1shRNA complimentary to the sequence responsible for mTNFR1 was also constructed and further confirmed by sequence analysis.To investigate the effect of mTNFR1shRNA plasmid on mTNFR1 expression in vivo and the disease progress in MHV-3 induced fulminant hepatitis mice model.By hydrodynamic injection of mTNFRlshRNA plasmid,the survival rate of mice,hepatic pathological change were examined and compared between mice with/without mTNFR1 shRNA plasmid intervention.The expression of mTNFR1 was detected by Real-time PCR,immunohistochemistry staining.The mTNFR1 shRNA plasmid significantly reduced mTNFR1 expression in vivo,markedly ameliorates inflammatory infiltration,prolonged the survival time period and elevated the survival rate from 0 up to 13.3% in Balb/cJ mice with MHV-3 induced fulminant hepatitis.This study was designed to explore the opportunity of RNA interference technique in inhibiting TNFR1 expression,which has been reported to be involved in the development of a variety of diseases including fulminant viral hepatitis and severe chronic hepatitis B.
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Objective To construct the eukaryotic expression plasmids of human Fas and TNFR1 gene(pcDNA3.0-hFas and pcDNA3.0-hTNFR1)and microRNA(miRNA)expression plasmid of hFas and hTNFR1 named p-hFasmiRNA and p-hTNFR1miRNA,and to investigate their inhibitory effects in vitro.Methods The eukaryotic expression plasmids of human Fas and TNFR1 gene were constructed(pcDNA3.0-hFas and pcDNA3.0-hTNFR1)and have been shown successfully to express hFas and hTNFR1 protein.miRNA expression plasmids of hFas and hTNFR1 named p-hFasmiRNA and p-hTNFR1miRNA complimentary to the sequence responsible for hFas and hTNFR1 respective were constructed,meanwhile irrelevant miRNA plasmid was used as a control.By respective co-transfection of p-hFasmiRNA and pcDNA3.0-hFas,p-hTNFR1 miRNA and pcDNA3.0-hTNFR1 expression construct into 293T cells,the inhibition of hFas and hTNFR1 expression was analyzed by real-time PCR and Western blot.Results The experiments showed the significant inhibitory effect of p-hFasmiRNA on hFas and p-hTNFR1miRNA on hTNFR1 expression at 48 h post-transfection both at RNA level and protein level as well in 293T cell lines with the inhibitory efficiency being as high as 87% for hFas and 80% for hTNFR1,respectively.Conclusion The p-hFasmiRNA and p-hTNFR1miRNA were constructed successfully,and it was verified that they could specifically inhibit the hFas and hTNFR1 expression at the cellular level.
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In Chagas disease, understanding how the immune response controls parasite growth but also leads to heart damage may provide insight into the design of new therapeutic strategies. Tumor necrosis factor-alpha (TNF-á) is important for resistance to acute Trypanosoma cruzi infection; however, in patients suffering from chronic T. cruzi infection, plasma TNF-á levels correlate with cardiomyopathy. Recent data suggest that CD8-enriched chagasic myocarditis formation involves CCR1/CCR5-mediated cell migration. Herein, the contribution of TNF-á, especially signaling through the receptor TNFR1/p55, to the pathophysiology of T. cruzi infection was evaluated with a focus on the development of myocarditis and heart dysfunction. Colombian strain-infected C57BL/6 mice had increased frequencies of TNFR1/p55+ and TNF-á+ splenocytes. Although TNFR1-/- mice exhibited reduced myocarditis in the absence of parasite burden, they succumbed to acute infection. Similar to C57BL/6 mice, Benznidazole-treated TNFR1-/- mice survived acute infection. In TNFR1-/- mice, reduced CD8-enriched myocarditis was associated with defective activation of CD44+CD62Llow/- and CCR5+ CD8+ lymphocytes. Also, anti-TNF-á treatment reduced the frequency of CD8+CCR5+ circulating cells and myocarditis, though parasite load was unaltered in infected C3H/HeJ mice. TNFR1-/- and anti-TNF-á-treated infected mice showed regular expression of connexin-43 and reduced fibronectin deposition, respectively. Furthermore, anti-TNF-á treatment resulted in lower levels of CK-MB, a cardiomyocyte lesion marker. Our results suggest that TNF/TNFR1 signaling promotes CD8-enriched myocarditis formation and heart tissue damage, implicating the TNF/TNFR1 signaling pathway as a potential therapeutic target for control of T. cruzi-elicited cardiomyopathy.
Subject(s)
Animals , Female , Mice , Anti-Inflammatory Agents/pharmacology , Antibodies, Monoclonal/pharmacology , /immunology , Chagas Cardiomyopathy/immunology , /immunology , Receptors, Tumor Necrosis Factor, Type I/antagonists & inhibitors , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Cell Movement , Chronic Disease , Chagas Cardiomyopathy/drug therapy , Flow Cytometry , Immunohistochemistry , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Receptors, Tumor Necrosis Factor, Type I/blood , Receptors, Tumor Necrosis Factor, Type I/immunology , Signal Transduction , Tumor Necrosis Factor-alpha/blood , Tumor Necrosis Factor-alpha/immunologyABSTRACT
The expression of silience of death domains (SODD) and its clinical significance and relationship with phospho-NF-κB-p65 proteins in bone marrow cells of childhood acute lymphoblastic leukaemia (ALL) were explored, and the expression of SODD and phospho-NF-κB-p65 in Jurkat cells treated with chemotherapeutic drugs was detected in order to find a new chemotherapeutic target. The expression of SODD and phospho-NF-κB-p65 proteins in bone marrow cells was detected by immunohistochemistry in 25 children with ALL. The apoptosis rate was measured by Annexin-V-Fluorescence/PI double-labeling flow cytometry and the expression of SODD and phospho-NF-κB-p65 proteins determined by Western blotting in the Jurkat cells. It was found that the expression of SODD and active P65 in ALL was significantly higher than that in normal control group (P＜0.05). The expression of the SODD and phospho-NF-κB-p65 proteins in the high-risk (HR) group was significantly higher than that in the standard-risk (SR) group (P＜0.05). The Pearson rank correlation analysis revealed that there was a positive correlation between SODD and phospho-NF-κB-p65 expression (P＜0.01, r=0.69). VCR could effectively induce the apoptosis of Jurkat cells, and down-regulate the expression of SODD and phospho-NF-κB-p65 proteins in a time-dependent manner, but DNR could not down-regulate the expression of SODD effectively. It was concluded that SODD may be closely related to the clinical classification and prognosis of ALL in children. The expression of SODD and phospho-NF-κB-p65 had a definite synergistic relationship with the onset and development of ALL. VCR could down-regulate the expression of SODD and inhibit the NF-κB activation, which could recover the sensibility of apoptosis in leukemic cells.
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0.05),and TRAF2 was related to the tumor grade only(P
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Objective:To study e ects of Saikosaponin a(SSa) on tumour necrosis factor-alpha(TNF-?) release and its receptor expression in cultured hippocampal astrocytes induced by pentylenetetrazol(PTZ).Methods:The in vitro cultured primary hippocampal astrocytes were randomly divided into control group(group A),PTZ-induced group(group B)PTZ10mmol/L+SSa groups(group C and group D,the SSa concentrations were 1.25mg/L and 0.625mg/L respectively).The extracellular uid TNF-? level and the expression of tumour necrosis factor receptor type 1(TNFR1) in hippocampal astrocytes were respectively detected by ELISA and Western-blot after PTZ-induced 2 hours.Results:the TNF-? level and TNFR1 expression of group B were signi cantly higher than that of group A,group C and group D(P
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Tumor necrosis factor (TNF) -alpha induces pleiotropic cellular effects through a 55kDa, type 1 receptor (TNFR1) and a 75kDa type 2 receptor (TNFR2). Moreover, it participates in the pathogenesis of several CNS diseases, including demyelinating diseases. TNF- receptors are differentially expressed and are regulated in many cell types. However, data regarding the TNF-alpha receptor expression and regulation in human astrocytes is limited to date. We investigated TNF-alpha receptor expression, its regulation by cytokines, and its functional role in primary cultured human fetal astrocytes, which are the most abundant cellular population in the central nervous system and are known to be immunologically active. In this study, astrocytes were found to constitutively and predominantly transcribe, translate and shed TNFR1 rather than TNFR2, but TNFR2 expression was increased by adding TNF-alpha, IL-1, and IFN-gamma, but not by adding LPS. To determine the functional roles of TNFR1 and TNFR2 on TNF induction, we investigated NF-kappaB activation and TNF-alpha induction after neutralizing TNFR1 and TNFR2 by an antibody treatment. We found that NF-kappaB activation and TNF-alpha induction are blocked by TNFR1 neutralizing antibody treatments.
Subject(s)
Humans , Receptors, Tumor Necrosis Factor, Type II/genetics , Receptors, Tumor Necrosis Factor, Type I/genetics , RNA, Messenger/metabolism , NF-kappa B/metabolism , Gene Expression Regulation , Fetus/cytology , Cytokines/pharmacology , Cells, Cultured , Astrocytes/drug effectsABSTRACT
Objective:Comparing the expression of tumor necrosis factor receptor 1 (TNFR1) in decidua tissue and soluble tumor necrosis factor receptor 1(sTNFR1) in serum of normal pregnancy and spontaneous abortion mice to probe the relationship between TNFR1 and unexplained spontaneous abortion.Methods:The abortion-prone CBA?DBA/2 mating was established as the model of spontaneous abortion and nonabortion-prone CBA?BALB/c matings were used as the model of normal pregnancy.Immunohistochemistry method(SABC) was employed to detect the expression of TNFR1 in decidua tissue at the day 9 of gestation.The level of sTNFR1 in serum at the same time was determined by ABC-ELISA.Results:Compared with normal pregnancy model,the expression of TNFR1 in decidua tissue of spontaneous abortion was significantly increased (P