Objective To analyze the relationship between tumor necrosis factor-alpha (TNFα) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFα in peripheral blood mononuclear ce...Objective To analyze the relationship between tumor necrosis factor-alpha (TNFα) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFα in peripheral blood mononuclear cells (PBMC) in MG patients. Methods A biallelic polymorphism at position -308 in the promoter of TNFα gene was screened by PCR amplification and NcoI recognition site. One hundred and twenty-three MG cases and 115 healthy controls were included in this study. MG patients were classified to different groups according to clinical type, age at onset, and sex respectively. PBMC were isolated from 20 patients and 20 healthy controls, and then cultured in the presence or absence of phytohemag- glutinin (PHA) and acetycholine receptors (AchR). The supernatants were harvested after incubation and stored until TNFα was assayed by enzyme-linked immunosorbent assay. Results The frequency of TNFα-308 allele 2 (A) was found significantly increase in MG patients and showed a trend especially in late onset (≥ 40 years) and male patients (P < 0.05). The allele A had no relationship with thymic pathogenesis in MG patients. But frequency of allele A was significantly higher in general type than in ocular type (P < 0.05). MG patients had a higher inducible level of TNFα by PHA and AchR, and could be down regulated after treatment. Conclusion Polymorphism in TNFα gene promoter -308 is associated with onset of MG. The microsatellite allele TNFα2 confer risk for the development of MG in Chinese patients. MG patients have a higher inducible level of TNFα.展开更多
As a potent pleiotropic cytokine, tumor necrosis factor-alpha （TNF-ct） plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene （AjTNF-c0 in the Japanese eel （...As a potent pleiotropic cytokine, tumor necrosis factor-alpha （TNF-ct） plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene （AjTNF-c0 in the Japanese eel （Anguilla japonica） were identified and characterized. The full-length AjTNF-α cDNA was 1 546 bp, including a 5＇-untranslated region （UTR） of 13 bp, a 3＇-UTR of 879 bp and an open reading frame of 654 bp encoding a protein of 218 amino acids. The full-length genomic sequence of AjTNF-ct was 2 392 bp and included four exons and three introns. The putative AjTNF-α protein contained TNF family signature motifs, including a protease cleavage site, a transmembrane domain and two conserved cysteine residues. Quantitative real-time reverse transcription PCR analysis revealed AjTNF-α expression in a wide range of tissues, with predominant expression in blood and liver. Lower levels of expression were seen in spleen, gills, kidney, intestine, heart, and skin, with very low levels in muscle. The modulation of AjTNF-ct expression after injection of eels with lipopolysaccharide （LPS）, the viral mimic, poly I：C, or Aeromonas hydrophila was assessed in blood, liver, and kidney. In blood, TNF-α mRNA levels increased rapidly and then rapidly decreased after stimulation with LPS, poly I：C or A. hydrophila. However, the response to LPS and A. hydrophila peaked at 6 h while for poly I：C the peak was at 12 h. In liver, after injection with A. hydrophila, an up- and down-regulation of AjTNF-ct expression occurred twice, peaking at 6 h and 24 h, respectively. No remarkable increase of AjTNF-α expression appeared in liver until 72 h after LPS or poly I：C treatment. In kidney, ApiNF-α expression increased significantly only at 72 h post-stimulation with LPS orA. hydrophila. Our results suggest that AjTNF-α plays an important role in fish in the defense against viral and bacterial infection.展开更多
Objective To investigate the effect of peroxisome proliferator-activated receptor-α(PPARα) and PPARγactivators on tumor necrosis factor-α(TNFα) expression in neonatal rat cardiac myocytes. Methods Primary culture...Objective To investigate the effect of peroxisome proliferator-activated receptor-α(PPARα) and PPARγactivators on tumor necrosis factor-α(TNFα) expression in neonatal rat cardiac myocytes. Methods Primary cultures of cardiac myocytes from 1- to 3-day-old Wistar rats were prepared, and myocytes were ex-posed to lipopolysaccharide (LPS) and varying concentrations of PPARαor PPARγactivator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγexpression in cultured cardiac myocytes. Transient tr-ansfection of TNFαpromoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed. Results Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFαmRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARαor PPARγmRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFαpromoter activity was observed when myocytes was transiently transfected with whole length of TNFαpromoter (-721/+17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFαreporter construct in deletion of NF-κB binding site (-182/+17). Conclusions PPARαand PPARγactivators may inhibit cardiac TNFαexpression but not accompanied by change of PPARαor PPARγmRNA expression. Therefore PPARαand PPARγactivators appear to play a role in anti-inflammation. The mechanism may partly be involved in suppression of the NF-κB pathway.展开更多
This study was designed to systematically investigate expression of TNF α,IL-1β,I1-6 mRNA in the liver,lungs and kidneys, release of TNF α in the above tissues,their relationship with hepatic, pulmonary and renal d...This study was designed to systematically investigate expression of TNF α,IL-1β,I1-6 mRNA in the liver,lungs and kidneys, release of TNF α in the above tissues,their relationship with hepatic, pulmonary and renal dysfunction,and distribution of endogenous endotoxin in tissues after hemorrhagic shock in mice and rats, with reverse-transcription-polymerase chain reaction, ELISA, etc, to elucidate the kinetics of expression and release of major cytokines in vital organs,their role and mechanism of production in shock. The results were: ①expression of TNF α, IL-1 β, IL-6 mRNA in vital organs successively increased after hemorrhagic shock and resuscitation,and TNF α expression was the first to appear followed by IL-1β. Though expression of IL-6 mRNA appeared late,it persisted longer;②TNF α levels in the liver,lungs and kidneys were all elevated but to different degrees after shock and resuscitation. At 3 hours after resuscitation,TNF α levels in the three above tissues were still significantly high,while plasma TNF α levels were already decreased to control levels; ③hepatic,pulmonary and renal functions were damaged to different degrees after hemorrhagic shock,with hepatic dysfunction being the most severe; ④endotoxin levels in the liver, lungs and kidneys were markedly increased after shock and resuscitation,and paraIleled the expression of cytokine genes. In addition,there was slgniflcant correlatlon between changes in endotoxin level in tissues and TNF α release in tissues during early shock. lt is suggested that expression and release of cytokines in vital organs might play an important role in local organ damage after hemorrhagic shock,and production of cytokines is related to endotoxin translocation.展开更多
Objective.In this study,we investigated the hypothesis that tumor necrosis factor(TNF)α-308gene polymorphism might be of the genetic predisposition to asthma and asthma phenotypes.Methods.TNFα-308gene polymorphism w...Objective.In this study,we investigated the hypothesis that tumor necrosis factor(TNF)α-308gene polymorphism might be of the genetic predisposition to asthma and asthma phenotypes.Methods.TNFα-308gene polymorphism was genotyped in221random unrelated Northern Chinese population(comprising125asthmatics and96healthy controls)and52individuals from12asthmatic families with Han ethnic by using polymerase chain reaction(PCR)-restriction fragment length polymor-phism(RFLP).Methacholine(Mch)broncho-challenge test,bronchial reversibility test and lung function were underwent in all asthmatics.Results.TNFα-3082homozygosity was present at a significantly higher frequency in asthmatics than that in controls(20.8%vs11.4%,P<0.05,OR2.259),the TNF allele2was also higher in asthmatics compared with controls(0.42vs0.33,P<0.01).TNFα-3082homozygosity was an weak independent risk factor for asthma etiology(OR0.226,P<0.05).Moreover,patients carrying TNFα-3082homozy-gosity had less responsive to inhaledβ 2 -agonist in20minutes than patients carrying other two genotypes(24.1%vs29.5%vs38.8%,P<0.05).Linkage analysis didn’t support that TNFαgene was linked to asthma (Likelihood of odds,LOD<1)based on familial data.Conclusion These results suggest that TNFα-3082homozygosity may be of a component contribut-ing to the genetic predisposition to asthma ,and airway responsiveness toβ2 -agonist.展开更多
文摘Objective To analyze the relationship between tumor necrosis factor-alpha (TNFα) gene promoter -308 polymorphism and myasthenia gravis (MG) in Chinese and analyze secretion of TNFα in peripheral blood mononuclear cells (PBMC) in MG patients. Methods A biallelic polymorphism at position -308 in the promoter of TNFα gene was screened by PCR amplification and NcoI recognition site. One hundred and twenty-three MG cases and 115 healthy controls were included in this study. MG patients were classified to different groups according to clinical type, age at onset, and sex respectively. PBMC were isolated from 20 patients and 20 healthy controls, and then cultured in the presence or absence of phytohemag- glutinin (PHA) and acetycholine receptors (AchR). The supernatants were harvested after incubation and stored until TNFα was assayed by enzyme-linked immunosorbent assay. Results The frequency of TNFα-308 allele 2 (A) was found significantly increase in MG patients and showed a trend especially in late onset (≥ 40 years) and male patients (P < 0.05). The allele A had no relationship with thymic pathogenesis in MG patients. But frequency of allele A was significantly higher in general type than in ocular type (P < 0.05). MG patients had a higher inducible level of TNFα by PHA and AchR, and could be down regulated after treatment. Conclusion Polymorphism in TNFα gene promoter -308 is associated with onset of MG. The microsatellite allele TNFα2 confer risk for the development of MG in Chinese patients. MG patients have a higher inducible level of TNFα.
基金Supported by the Ocean and Fishery Bureau of Fujian Province(No.201212140006)the Scientific Research Foundation of Jimei University,China(No.ZQ2008013)+1 种基金the National Natural Science Foundation of China(No.31272685)the Program of the Education Department of Fujian Province(No.JA11150)
文摘As a potent pleiotropic cytokine, tumor necrosis factor-alpha （TNF-ct） plays an important role in innate immune responses. The cDNA sequence and genomic structure of the TNF-α gene （AjTNF-c0 in the Japanese eel （Anguilla japonica） were identified and characterized. The full-length AjTNF-α cDNA was 1 546 bp, including a 5＇-untranslated region （UTR） of 13 bp, a 3＇-UTR of 879 bp and an open reading frame of 654 bp encoding a protein of 218 amino acids. The full-length genomic sequence of AjTNF-ct was 2 392 bp and included four exons and three introns. The putative AjTNF-α protein contained TNF family signature motifs, including a protease cleavage site, a transmembrane domain and two conserved cysteine residues. Quantitative real-time reverse transcription PCR analysis revealed AjTNF-α expression in a wide range of tissues, with predominant expression in blood and liver. Lower levels of expression were seen in spleen, gills, kidney, intestine, heart, and skin, with very low levels in muscle. The modulation of AjTNF-ct expression after injection of eels with lipopolysaccharide （LPS）, the viral mimic, poly I：C, or Aeromonas hydrophila was assessed in blood, liver, and kidney. In blood, TNF-α mRNA levels increased rapidly and then rapidly decreased after stimulation with LPS, poly I：C or A. hydrophila. However, the response to LPS and A. hydrophila peaked at 6 h while for poly I：C the peak was at 12 h. In liver, after injection with A. hydrophila, an up- and down-regulation of AjTNF-ct expression occurred twice, peaking at 6 h and 24 h, respectively. No remarkable increase of AjTNF-α expression appeared in liver until 72 h after LPS or poly I：C treatment. In kidney, ApiNF-α expression increased significantly only at 72 h post-stimulation with LPS orA. hydrophila. Our results suggest that AjTNF-α plays an important role in fish in the defense against viral and bacterial infection.
基金Supported by the National Nature Science Foundation of China (30270551) and Military "10.5"Foundation (02M012).
文摘Objective To investigate the effect of peroxisome proliferator-activated receptor-α(PPARα) and PPARγactivators on tumor necrosis factor-α(TNFα) expression in neonatal rat cardiac myocytes. Methods Primary cultures of cardiac myocytes from 1- to 3-day-old Wistar rats were prepared, and myocytes were ex-posed to lipopolysaccharide (LPS) and varying concentrations of PPARαor PPARγactivator (fenofibrate or pioglitazone).RT-PCR and ELISA were used to measure TNFα, PPARα, and PPARγexpression in cultured cardiac myocytes. Transient tr-ansfection of TNFαpromoter with or without nuclear factor-kappaB (NF-κB) binding site to cardiac myocytes was performed. Results Pretreatment of cardiac myocytes with fenofibrate or pioglitazone inhibited LPS-induced TNFαmRNA and protein expression in a dose-dependent manner. However, no significant changes were observed on PPARαor PPARγmRNA expression when cardiac myocytes were pretreated with fenofibrate or pioglitazone. Proportional suppression of TNFαpromoter activity was observed when myocytes was transiently transfected with whole length of TNFαpromoter (-721/+17) after being stimulated with LPS and fenofibrate or pioglitazone, whereas no change of promoter activity was observed with transfection of TNFαreporter construct in deletion of NF-κB binding site (-182/+17). Conclusions PPARαand PPARγactivators may inhibit cardiac TNFαexpression but not accompanied by change of PPARαor PPARγmRNA expression. Therefore PPARαand PPARγactivators appear to play a role in anti-inflammation. The mechanism may partly be involved in suppression of the NF-κB pathway.
文摘This study was designed to systematically investigate expression of TNF α,IL-1β,I1-6 mRNA in the liver,lungs and kidneys, release of TNF α in the above tissues,their relationship with hepatic, pulmonary and renal dysfunction,and distribution of endogenous endotoxin in tissues after hemorrhagic shock in mice and rats, with reverse-transcription-polymerase chain reaction, ELISA, etc, to elucidate the kinetics of expression and release of major cytokines in vital organs,their role and mechanism of production in shock. The results were: ①expression of TNF α, IL-1 β, IL-6 mRNA in vital organs successively increased after hemorrhagic shock and resuscitation,and TNF α expression was the first to appear followed by IL-1β. Though expression of IL-6 mRNA appeared late,it persisted longer;②TNF α levels in the liver,lungs and kidneys were all elevated but to different degrees after shock and resuscitation. At 3 hours after resuscitation,TNF α levels in the three above tissues were still significantly high,while plasma TNF α levels were already decreased to control levels; ③hepatic,pulmonary and renal functions were damaged to different degrees after hemorrhagic shock,with hepatic dysfunction being the most severe; ④endotoxin levels in the liver, lungs and kidneys were markedly increased after shock and resuscitation,and paraIleled the expression of cytokine genes. In addition,there was slgniflcant correlatlon between changes in endotoxin level in tissues and TNF α release in tissues during early shock. lt is suggested that expression and release of cytokines in vital organs might play an important role in local organ damage after hemorrhagic shock,and production of cytokines is related to endotoxin translocation.
文摘Objective.In this study,we investigated the hypothesis that tumor necrosis factor(TNF)α-308gene polymorphism might be of the genetic predisposition to asthma and asthma phenotypes.Methods.TNFα-308gene polymorphism was genotyped in221random unrelated Northern Chinese population(comprising125asthmatics and96healthy controls)and52individuals from12asthmatic families with Han ethnic by using polymerase chain reaction(PCR)-restriction fragment length polymor-phism(RFLP).Methacholine(Mch)broncho-challenge test,bronchial reversibility test and lung function were underwent in all asthmatics.Results.TNFα-3082homozygosity was present at a significantly higher frequency in asthmatics than that in controls(20.8%vs11.4%,P<0.05,OR2.259),the TNF allele2was also higher in asthmatics compared with controls(0.42vs0.33,P<0.01).TNFα-3082homozygosity was an weak independent risk factor for asthma etiology(OR0.226,P<0.05).Moreover,patients carrying TNFα-3082homozy-gosity had less responsive to inhaledβ 2 -agonist in20minutes than patients carrying other two genotypes(24.1%vs29.5%vs38.8%,P<0.05).Linkage analysis didn’t support that TNFαgene was linked to asthma (Likelihood of odds,LOD<1)based on familial data.Conclusion These results suggest that TNFα-3082homozygosity may be of a component contribut-ing to the genetic predisposition to asthma ,and airway responsiveness toβ2 -agonist.
