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1.
Inducible nitric oxide synthase (iNOS) protein was detected immunohistochemically in formalin-fixed, paraffin wax-embedded lung tissues from 10 natural cases of porcine pleuropneumonia. Positive cells typically exhibited a red reaction product without background staining. Labelling of iNOS protein was intense in "oat cells", the clustered leucocytes with streaming nuclear chromatin that are a characteristic histological feature of porcine pleuropneumonia. Macrophages and neutrophils within alveolar spaces but not within blood vessels consistently showed iNOS labelling, but such labelling was minimal in non-lesional lung of Actinobacillus pleuropneumoniae -infected pigs and in normal lung from control pigs. The results suggest that iNOS plays a role in pathophysiological processes during A. pleuropneumoniae infection.  相似文献   

2.
Cyclooxygenase-2 (COX-2) protein was detected immunohistochemically in formalin-fixed, paraffin wax-embedded lung tissues from 15 pigs with naturally occurring pleuropneumonia caused by Actinobacillus pleuropneumoniae. Positive cells typically exhibited a red reaction product without background staining. Alveolar macrophages, neutrophils, and bronchial and bronchiolar epithelial cells had positive immunohistochemical signals. Immunoreactivity of COX-2 protein was intense in the clustered leucocytes with streaming nuclear chromatin that are a characteristic histological feature of porcine pleuropneumonia. COX-2 protein was always associated with macrophages and neutrophils in pleuropneumonic lung lesions but was minimal in non-lesional lung of A. pleuropneumoniae -infected pigs and in normal lung from control pigs. The results suggest that COX-2 plays a role in pathophysiological processes during A. pleuropneumoniae infection.  相似文献   

3.
The apxIV gene was detected, by in-situ hybridization with a non-radioactive digoxigenin-labelled probe, in formalin-fixed, paraffin wax-embedded samples of lung tissue from 10 pigs naturally infected with Actinobacillus pleuropneumoniae. A 442 base pair DNA probe of the apxIV gene from A. pleuropneumoniae serotype 2 was generated by the polymerase chain reaction. All 10 pigs infected with A. pleuropneumoniae serotypes 2, 5, 6, or an untypable strain showed a distinct, positive signal in the degenerate alveolar leucocytes in alveolar spaces, and in the dense zone of degenerated cells in granulation tissue surrounding the necrotic areas. Thus, the study demonstrated the presence of the apxIV gene in pleuropneumonic lesions caused by A. pleuropneumoniae. Copyright Harcourt Publishers Ltd.  相似文献   

4.
5.
A rapid, simple, and accurate counterimmunoelectrophoresis technique was developed for serotyping cultures of Actinobacillus pleuropneumoniae as well as for detection of their type-specific antigens in the lung tissues of infected pigs. The counterimmunoelectrophoresis test correctly identified all of the reference antigens and more than 99% of 1,200 field isolates of A. pleuropneumoniae representing the 12 established serotypes within 1 h. Counterimmunoelectrophoresis and coagglutination tests did not differ broadly in sensitivity from each other. Both procedures were more rapid and more sensitive than immunodiffusion and indirect hemagglutination tests. A total of 355 lung tissue samples (130 lungs of pigs that died because of acute respiratory problems, 125 lungs of pigs from herds with chronically infected pleuropneumonia, and 100 lungs from apparently healthy pigs at the slaughterhouse) were examined for the presence of A. pleuropneumoniae type-specific antigens by counterimmunoelectrophoresis, coagglutination, and immunodiffusion tests. A. pleuropneumoniae type-specific antigen was found in all 55 samples from which the bacteria had earlier been isolated and in 27 specimens in which they had not been found. Detection of antigen in the lung tissues by coagglutination and counterimmunoelectrophoresis tests was found to be much simpler and much more rapid than conventional culture isolation. Both counterimmunoelectrophoresis and coagglutination tests were found extremely useful in the diagnosis of acute cases of porcine pleuropneumonia. However, these techniques were able to detect only some of the chronically infected carrier pigs.  相似文献   

6.
内皮型、诱导型一氧化氮合酶在乳腺癌中的表达   总被引:1,自引:0,他引:1  
目的 :研究内皮型一氧化氮合酶 (eNOS)、诱导型一氧化氮合酶 (iNOS)在乳癌中表达及与淋巴结转移的关系。方法 :采用免疫组化S P法检测 60例乳癌中eNOS和iNOS的表达。结果 :eNOS和iNOS阳性在乳癌中表达率分别为 75 0 %和71 7%。在淋巴结转移组和无淋巴结转移组中eNOS阳性表达率分别为 66 7%和 83 3 % ,两组间差异无统计学意义 (χ2 =2 2 2 ,P >0 0 5) ,而iNOS在淋巴结转移和无转移组中阳性表达率分别为 53 3 %和 90 0 % ,两组间差异有统计学意义 (χ2 =9 93 ,P <0 0 1 )。结论 :内皮型、诱导型一氧化氮合酶在乳腺癌中高表达 ;iNOS的表达与乳腺癌的淋巴转移相关  相似文献   

7.
We have recently established a murine model of pulmonary and disseminated infection with a highly virulent strain of Cryptococcus neoformans and demonstrated that administration of interleukin-12 (IL-12) protected the animals against infection. In this study, we extended these studies by investigating the host defense mechanisms. In particular, we examined the expression of mRNA for helper T-cell 1 (Th1) cytokines (IL-2, lymphotoxin, and gamma interferon [IFN-gamma]), Th2 cytokines (IL-4, -6, and -10), macrophage-derived cytokines (tumor necrosis factor alpha [TNF-alpha], IL-1beta, transforming growth factor beta [TGF-beta, IL-12p40, and IFN-gamma-inducing factor [IGIF]), and inducible nitric oxide synthase (iNOS) in the lungs on days 1, 3, 7, and 14 after infection and following treatment with IL-12. There was little or no expression of mRNAs for Th1 cytokines, TNF-alpha, IL-12p40, IGIF, and iNOS in the infected mice, but expression increased markedly after treatment with IL-12. In contrast, the mRNAs for Th2 cytokines, IL-1beta, and TGF-beta were detected at considerable levels during the early stages of infection, and, interestingly, expression was not suppressed by IL-12 but rather augmented, particularly during the late stage. Similar results were also obtained for IFN-gamma, IL-4, IL-10, and TNF-alpha measured in the lung homogenates by enzyme-linked immunosorbent assay. These results suggest that the predominance of expression of Th2 cytokines and TGF-beta over Th1 cytokines, TNF-alpha, IL-12p40, IGIF, and iNOS is associated with severe lethal infection in mice and that administration of IL-12 protects infected animals by stimulating Th1 cytokines.  相似文献   

8.
9.
Pressure-related activation of inducible nitric oxide synthase  相似文献   

10.
The detection and distribution of interleukin-1 (IL-1), tumour necrosis factor-alpha (TNF-alpha) and IL-6 were studied, by in-situ hybridization with a non-radioactive digoxigenin-labelled probe, in formalin-fixed paraffin wax- embedded lung tissue from 10 pigs naturally infected with Actinobacillus pleuropneumoniae. A strong hybridization signal for IL-1, TNF-alpha and IL-6 was detected in "streaming" degenerate alveolar leucocytes (the so-called "oat cells") bordering zones of coagulative necrosis, and a less intense signal was seen in the dense zone of degenerate cells in granulation tissue surrounding the necrotic areas. IL-1 expression was also prominent in scattered endothelial cells bordering zones of coagulative necrosis. Simultaneous expression of all three cytokines was always associated with pleuropneumonic lung lesions. Expression of inflammatory cytokines was minimal in non-lesional lung tissue of the infected pigs and in normal lung from control pigs. The results suggest that these cytokines play a crucial role in mediating and regulating inflammation through cells of several types in A. pleuropneumoniae infection. 1999 Harcourt Publishers Ltd.  相似文献   

11.
 目的:探讨一氧化氮(NO)/诱导型一氧化氮合酶(iNOS)在动脉粥样硬化(atherosclerosis,AS)过程中的动态变化,分析其对动脉粥样硬化形成过程的影响。方法:将60只SD大鼠随机分成2组:对照组及AS组,每组30只。AS组采用维生素D3腹腔注射联合高脂饲料饲养的方法构建动脉粥样硬化模型。用相关生化方法检测血清各项生化指标:总胆固醇、甘油三酯、高密度脂蛋白胆固醇、低密度脂蛋白胆固醇、空腹血糖和钙离子,比色法检测血清NO浓度,并对主动脉行HE染色,免疫组化技术检测iNOS蛋白表达,将所得数据进行统计分析,用简单线性相关分析NO与钙离子及动脉粥样硬化指数的相关性。结果:90 d后成功构建了主动脉中膜钙化型动脉粥样硬化模型。血清NO浓度在动脉粥样硬化过程中逐步下降,各组间差异均有统计学意义(均P<0.05)。动脉粥样硬化过程中动脉粥样硬化指数与钙离子呈正相关,与NO呈负相关。在90 d的AS组粥样斑块区免疫组化技术检测到iNOS蛋白表达。结论:在动脉粥样硬化形成过程中,主动脉粥样斑块区iNOS蛋白高表达,但血清NO浓度逐渐降低,NO抗动脉粥样硬化作用减弱。  相似文献   

12.
目的:探讨NF-κB的活性及iNOS基因表达在低氧性肺动脉高压(HPH)发病过程中的变化。方法:复制低氧性肺动脉高压大鼠模型,用免疫组化、原位杂交、半定量逆转录-聚合酶链式反应(RT-PCR)和Western blot等方法进行检测。结果:iNOS mRNA在腺泡内肺动脉(IAPA)的表达,低氧28 d(H28d)组染色强于正常(N)组、低氧5 d(H5d)组和低氧14 d(H14d)组。半定量RT-PCR证实低氧肺组织iNOS mRNA含量在H28d组分别是N组、H5d组和H14d组的2.1倍、1.9倍和1.8倍。H28d组肺组织NF-κB的核染色增多,I-κBα的含量在N组、H5d组和H14d组分别是H28d组的2.7倍、2.8倍和2.5倍。结论:在HPH中NF-κB的激活可能与低氧肺血管构建及iNOS mRNA的表达有关。  相似文献   

13.
NF-κB的活性和iNOS基因表达在低氧性肺动脉高压中的变化   总被引:2,自引:0,他引:2  
目的:探讨NF-κB的活性及iNOS基因表达在低氧性肺动脉高压(HPH)发病过程中的变化. 方法: 复制低氧性肺动脉高压大鼠模型, 用免疫组化、原位杂交、半定量逆转录-聚合酶链式反应(RT-PCR)和Western blot等方法进行检测. 结果:iNOS mRNA在腺泡内肺动脉(IAPA)的表达,低氧28 d(H28 d)组染色强于正常(N)组、低氧5 d(H5 d)组和低氧14 d(H14 d)组.半定量RT-PCR证实低氧肺组织iNOS mRNA含量在H28 d组分别是N组、H5 d组和H14 d组的2.1倍、1.9倍和1.8倍.H28 d组肺组织NF-κB的核染色增多,I-κBa的含量在N组、H5 d组和H14 d组分别是H28 d组的2.7倍、2.8倍和2.5倍. 结论: 在HPH中NF-κB的激活可能与低氧肺血管构建及iNOS mRNA的表达有关.  相似文献   

14.
The present study was designed to elucidate the dynamic changes of nitric oxide (NO) production in the perilymph and to investigate the immunostaining for inducible nitric oxide synthase (iNOS) in the cochlea for 7 days after transient cochlear ischemia. Moreover, aminoguanidine, which is a selective iNOS inhibitor, was administrated immediately following ischemia and every 24h thereafter for 7 days to investigate whether the production of NO is dependent on the iNOS pathway. Significant increases in the oxidative NO metabolites, nitrite (NO(2)(-)) and nitrate (NO(3)(-)), were measured on day 1 using an in vivo microdialysis and on-line high performance liquid chromatography (HPLC) system. The immunostaining for iNOS was strongly expressed on days 1 and 4 and returned to normal on day 7 after the ischemia. The administration of aminoguanidine reduced the oxidative NO metabolites on day 1 and suppressed the expression of iNOS. These findings suggest that transient ischemia causes a remarkable increase in NO production in the perilymph, which might be attributable to the iNOS pathway.  相似文献   

15.
16.
The capsular polysaccharide (CP) of Actinobacillus pleuropneumoniae is required for virulence of the bacteria in swine. However, a molecular investigation of whether the type or quantity of CP affects A. pleuropneumoniae virulence has not been reported. To initiate this investigation, a DNA region downstream of conserved genes required for CP export in A. pleuropneumoniae serotype 1 was cloned and sequenced. Three open reading frames, designated cps1A, cps1B, and cps1C, were identified that had amino acid homology to bacterial carbohydrate biosynthesis genes. A kanamycin resistance cassette (Kan(r)) was inserted into a 750-bp deletion spanning cps1AB or into a 512-bp deletion in cps1B only, and the constructs were cloned in a suicide vector. The Kan(r) gene was then transferred into the chromosome of strain 4074 by homologous recombination to produce strain 4074Deltacps1N and strain 4074Deltacps1B, respectively. Strain 4074Deltacps1N produced no detectable CP, but strain 4074Deltacps1B made 15% of the serotype 1 CP made by the parent strain, 4074, as determined by enzyme-linked immunosorbent assay and precipitation of free CP. The cps1ABC genes of strain 4074 and the cps5ABC and cps5ABCDE genes of serotype 5a strain J45 were cloned into the shuttle vector pLS88 and electroporated into 4074Deltacps1N to produce 4074Deltacps1N(pABcps101), 4074Deltacps1N(pJMLcps53), and 4074Deltacps1N(pABcps55), respectively. Strain 4074Deltacps1N(pABcps101) produced about 33% of the serotype 1 CP produced by strain 4074. Strains 4074Deltacps1N(pJMLcps53) and 4074Deltacps1N(pABcps55) produced serotype 5a CP in similar quantity or in fourfold excess, respectively, to that produced by strain 4074. With intratracheal challenge in pigs at similar dosages, the order of virulence of strains producing serotype 1 CP (assessed by mortality, lung consolidation, hemorrhage, and fibrinous pleuritis) was the following: strain 4074 > strain 4074Deltacps1N(pABcps101) > or = strain 4074Deltacps1N > strain 4074Deltacps1B. Strain 4074Deltacps1N(pJMLcps53) was less virulent than strain 4074Deltacps1N(pABcps55). However, both strains produced serotype 5a CP in similar or greater quantities than was observed for production of serotype 1 CP by the parent strain, 4074, but were less virulent than the parent strain. Therefore, the amount of serotype 1 or 5a CP produced by isogenic strains of A. pleuropneumoniae correlated with the virulence of the bacteria in pigs. However, virulence was also influenced by the type of CP produced or by its mechanism of expression.  相似文献   

17.
BACKGROUND: The deficiency of the inducible nitric oxide synthase (iNOS) substrate, L-arginine (L-Arg), the co-factor tetrahydrobiopterin (H4B) or molecular oxygen may lead to lower NO levels, which enhances the development of adhesion phenotype. METHODS: We utilized high-performance liquid chromatography (HPLC) and immunoprecipitation with nitrotyrosine antibody to determine the levels of H4B, citrulline and protein nitration in fibroblasts established from normal peritoneal and adhesion tissues. RESULTS: The level of H4B was dramatically attenuated in adhesion fibroblasts. The immunoprecipitation with nitrotyrosine antibody revealed higher protein nitration in adhesion compared with normal fibroblasts. There were higher accumulations of citrulline in adhesion fibroblasts as compared with normal fibroblasts. In addition, peritoneal fibroblasts treated with 2% oxygen for 24 h and implanted back into the peritoneal cavity of the rats exhibited marked increase in severity of adhesion as well as extensive distribution involving many sites and organs. CONCLUSIONS: Control of the catalytic activity of iNOS in adhesion fibroblasts may be because of subsaturating amounts of L-Arg and H4B which allow iNOS to generate a combination of reactive oxygen species in addition to NO, thereby influencing NO bioavailability and function.  相似文献   

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目的:探讨椎基底动脉缺血再灌注耳蜗组织损伤方式及可能的损伤机制。方法:健康豚鼠72只,随机分成正常组、假手术组、模型组,每组各8只。用组织化学方法观察缺血再灌注不同时间耳蜗各部位的组织改变;用免疫组织化学法(ABC)检测各组动物耳蜗中诱导型一氧化氮合酶的表达和变化;用原位凋亡法(TUNEL法)观察耳蜗的细胞凋亡情况。结果:缺血再灌注的内外毛细胞变形缺损、血管纹变薄等;诱导型一氧化氮合酶在缺血及再灌注的表达增强;正常组及假手术组没有或仅有个别部位出现细胞凋亡,缺血及再灌注各组内外毛细胞及螺旋神经节部位凋亡细胞明显增多。结论:再灌注期间细胞凋亡数较缺血期间明显增多,其原因为包括一氧化氮在内的多种氧自由基表达增高所致。  相似文献   

20.
Preeclampsia (PE) is a disease that onsets in the second half of pregnancy. This condition is characterized by hypertension, proteinuria and, frequently, intrauterine growth restriction (IUGR). Nitric oxide (NO) regulates blood flow in the human placenta, it induces vasodilatation, inhibition of platelet aggregation and prevents adhesion of platelets to endothelial cells. In this work, nitrite levels were evaluated in the sera of peripheral blood of normal pregnant women (n = 46) and women with PE (n = 50); additionally, the expression of endothelial constitutive nitric oxide and inducible synthases (eNOS and iNOS, respectively) of placental tissues, were determined. An increased concentration of serum nitrites from patients with PE, in relation to normal pregnant women (150.64 +/- 8.94 vs 40.62 +/- 1.65 microM, p < 0.00001) was observed. An increased expression of nitric oxide synthases (eNOS and iNOS), in the placental tissues of (PE) patients, as compared to that of normal pregnant women (iNOS 4.29 +/- 1.51 vs 0.59 +/- 0.13; eNOS 1.78 +/- 0.74 vs 0.46 +/- 0.22, p < 0.005) was also observed. Our results show that there exists a relationship between serum nitrites concentration and the expression of eNOS and iNOS, as analyzed in protein extracts of placental tissues.  相似文献   

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