哮喘患者外周血肿瘤坏死因子—α水平及影响因素

测定了２０例哮喘患者及１５例正常人的外周血肿瘤坏死因子－α（ＴＮＦ－α）水平，并分离哮喘患者外周血单个核细胞（ＰＢＭＮＣ）于体外加药培养，结果显示，哮喘患者外周血ＴＮＦ－α水平明显高于正常人，其在哮喘发病中具有重要意义，植物血凝素（ＰＨＡ），抗原可激发ＰＢＭＮＣ产生ＴＮＦ－α，治疗浓度氨茶碱可抑制ＰＢＭＮＣ分泌ＴＮＦ－α。     

脂多糖和抗炎药物对肺血管内巨噬细胞核因子κB的影响

目的观察脂多糖（ＬＰＳ）致肺血管内巨噬细胞（ＰＩＭ）核因子κＢ（ＮＦκＢ）的活化及抗炎药物地塞米松（ＤＥＸ）和阿斯匹林（ＡＳＡ）对ＮＦκＢ的影响。方法用改良法分离、培养猪ＰＩＭ,设正常对照、ＬＰＳ刺激、ＤＥＸ和ＡＳＡ干预组,共４组。用凝胶电泳迁移率改变分析（ＥＭＳＡ）法和放射免疫分析（ＲＩＡ）法,分别检测ＰＩＭ核提取物ＮＦκＢ的活性和细胞培养上清肿瘤坏死因子α（ＴＮＦα）的含量。结果ＬＰＳ刺激组ＮＦκＢ活性于刺激后０５～４小时、ＴＮＦα含量于刺激后１～２小时高于刺激前和正常对照组（Ｐ＜０．０１）;二者在刺激后１小时呈显著正相关（ｒ＝０．９９１,Ｐ＜０．０１）。ＤＥＸ组和ＡＳＡ组ＮＦκＢ活性、ＴＮＦα含量虽较刺激前和正常对照组有所升高,但均显著低于ＬＰＳ组（Ｐ＜０．０１）。结论ＬＰＳ可诱导ＰＩＭＮＦκＢ激活,并进而导致ＴＮＦα的基因转录和表达增加;ＤＥＸ和ＡＳＡ可通过抑制ＮＦκＢ的活化而减少ＴＮＦα的释放。     

肾小管和肿瘤坏死因子与间质纤维化的关系

目的：研究肾小管和肿瘤坏死因子与肾间质纤维化的关系。方法：肾小管上皮细胞及肾成纤维细胞的体外培养，采用逆转录－多聚酶链反应（ＲＴ－ＰＣＲ），免疫组化染色、放射免疫测定（ＲＩＡ）及双重免疫化染色技术，并测定＾３Ｈ－ＴｄＲ掺入率，观察ＴＮＦａ与肾成纤维细胞增殖的关系。结果：肾小管上皮细胞既有ＴＮＦａ－ｍＲＮＡ的表达，又有ＴＮＦ－Ｒ１ｍＲＮＡ的表达，还有ＴＮＦα的蛋白合成及分泌。且肾小管损伤及再生过程中     

TNF—α,IL—6和IL—8与慢性肝病的关系

为研究肿瘤坏死因子－α（ＴＮＦ－α）、白细胞介素－６（和白细胞介素－８（ＩＬ－８）与慢性肝病的关系，分别用ＲＩＡ和ＥＬＩＳＡ法检测９４例慢性肝病患者表ＴＮＦ－α和ＩＬ－８含量，同时用免疫组化ＡＢＣ法观察其中２５例患者（慢迁肝３例，慢活肝５例，肝硬变１７例）肝组织内ＴＮＦα，ＩＬ－８和ＩＬ－６的表达和分布，结果显示，慢活肝和肝硬变患者血清及肝组织内上述细胞因子水平均明显增加（Ｐ〈０．０５－０．０１）     

糖皮质激素对非IgA系膜增殖性肾炎患者肿瘤坏死因子的影响

目的观察非ＩｇＡ系膜增殖性肾炎患者应用激素前后血清肿瘤坏死因子－α（ＴＮＦ－α）的变化及其与尿蛋白的相关关系。方法３０例非ＩｇＡ系膜增殖性肾炎表现为肾病综合征的患者按病理增生程度不同分为轻度增生组和中重度增生组，两组均在用激素前及用激素后４周及８周测定血清ＴＮＦ－α及２４小时尿蛋白定量。结果两组患者治疗前血清ＴＮＦ－α明显升高，且ＴＮＦ－α水平与系膜增生程度有关，激素治疗后ＴＮＦ－α下降，疗效与系膜增生程度及ＴＮＦ－α水平有关。ＴＮＦ－α与２４小时尿蛋白存在显著正相关。结论对非ＩｇＡ系膜增殖性肾炎患者测定血清ＴＮＦ－α可作为判断组织学损伤的非创伤性参考指标及判断激素疗效与预后的参考指标。     

过敏性哮喘支气管肺泡灌洗液中肿瘤坏死因子－α等活性变化的研究

目的探讨肿瘤坏死因子－α（ＴＮＦ－α）、白细胞介素－６（ＩＬ－６）在哮喘中的意义。方法以卵白蛋白（ＯＡ）致敏和吸入激发复制哮喘豚鼠模型，以Ｌ９２９细胞杀伤和ＭＨ６０．ＢＳＦ２细胞增殖反应检测ＴＮＦ－α，ＩＬ－６生物活性。结果哮喘豚鼠支气管肺泡灌洗液（ＢＡＬＦ）及肺泡巨噬细胞（ＡＭ）上清液中ＴＮＦ－α、ＩＬ－６明显升高并呈动态改变。结论ＴＮＦ－α、ＩＬ－６可能在哮喘发生中起重要作用。     

内皮素肿瘤坏死因子在哮喘发病中的作用

为了探索内皮素（ＥＴ）、肿瘤坏死因子（ＴＮＦ－α）在哮喘中的作用及相互关系。实验测定了哮喘豚鼠血以及肺泡灌洗液（ＢＡＬＦ）中的内皮素－１（ＥＴ－１）、ＴＮＦ－α的释放量，以及离体气道平滑肌细胞在ＴＮＦ－α刺激下对ＥＴ－１释放的影响。观察到ＥＴ－１、ＴＮＦ－α的含量在哮喘组明显高于对照组，同时，ＴＮＦ－α可刺激平滑肌细胞分泌ＥＴ－１。表明ＴＮＦ－α引起的ＥＴ－１释放对气道高反应性的形成和气道重建可能     

肺纤维化大鼠肺泡巨噬细胞释放肿瘤坏死因子—α和血小板…

目的 动态观察博莱霉素（ＢＬＭ）致大鼠肺纤维化过程中肺泡巨噬细胞（ＡＭ）释放肿瘤坏死因子－α－（ＴＮＦ－α）和血小板源生长因子（ＰＤＧＦ）的变化。方法 采用ＥＬＩＳＡ法测定ＴＮＦ－α；采用生物性法测定ＰＤＧＦ。结果 （１）ＡＭ释放ＴＮＦ－α于灌注ＢＬＭ后第３天开始升高，第７天达高峰，１４天后虽有下降，但仍显著高于对照组；（２）ＡＭ释放的ＰＤＧＦ于灌注ＢＬＭ后第３天升高，第７天达高峰，第１４天后下降     

北冬虫夏草多糖对人肿瘤坏死因子α、白细胞介素2及其可溶性受体水平的影响

研究北冬虫夏草多糖对人外周血单个核细胞（ＰＢＭＣ）分泌肿瘤坏死因子α（ＴＮＦ－α），生成可溶性白细胞介素２受体（ｓＩＬ－２Ｒ）及人外周血淋巴细胞（ＰＢＬ）分泌白细胞介素２（ＩＬ－２）的影响。结果表明：北冬虫夏草多糖在１０～５０ｍｇ·Ｌ－１浓度时，刺激单个核细胞分泌ＴＮＦ－α（Ｐ＜０．０５）。在０．１～１０ｍｇ·Ｌ－１浓度时，刺激单个核细胞生成ｓＩＬ－２Ｒ（Ｐ＜０．０５，Ｐ＜０．０１），且在一定浓度范围内ＴＮＦ－α及ｓＩＬ－２Ｒ水平随北冬虫夏草多糖浓度增加而升高；北冬虫夏草多糖对刀豆蛋白Ａ（ＣｏｎＡ）刺激的人外周血淋巴细胞分泌ＩＬ－２水平有明显促进作用（Ｐ＜０．０５）。     

肿瘤坏死因子α在病毒性肝炎肝坏死中的作用

为探讨肿瘤坏死因子α（ＴＮＦα）在病毒性肝炎肝坏死中的作用，利用动物实验，对ＴＮＦα在肝坏死中的作用进行了研究。用抗ＴＮＦα单克隆抗体阻断由Ｄ－氨基半乳糖及细菌内毒素脂多糖所致的肝坏死；同时还以ＡＢＣ法，用抗ＴＮＦα单克隆抗体，对１３０例各型病毒性肝炎病人肝组织中ＴＮＦα的表达情况进行了研究。发现肝炎肝硬化、慢性重型肝炎及亚急性重型肝炎ＴＮＦα表达水平明显增加，ＴＮＦα表达水平与肝脏的炎症坏死程度有关。证实ＴＮＦα是病毒性肝炎肝坏死的重要介质。     

Ketamine suppresses intestinal NF-kappa B activation and proinflammatory cytokine in endotoxic rats

AIM: To investigate the protective effect of ketamine on the endotoxin-induced proinflammatory cytokines and NFkappa B activation in the intestine. ETHODS: Adult male Wistar rats were randomly divided into 6 groups: (a) normal saline control, (b) challenged with endotoxin (5 mg/kg) and treated by saline, (c) challenged with endotoxin (5 mg/kg) and treated by ketamine (0.5 mg/kg), (d) challenged with endotoxin (5 mg/kg) and treated by ketamine (5 mg/kg ), (e) challenged with endotoxin (5 mg/kg) and treated by ketamine (50 mg/kg), and (f) saline injected and treated by ketamine (50 mg/kg). After 1, 4 or 6 h, TNF-α and IL-6 mRNA were investigated in the tissues of the intestine (jejunum) by RT-PCR. TNF-α and IL-6 were measured by ELISA. We used electrophoretic mobility shift assay (EMSA) to investigate NF-kappa B activity in the intestine. RESULTS: NF-kappa B activity, the expression of TNF-α and IL-6 were enhanced in the intestine by endotoxin. Ketamine at a dose of 0.5 mg/kg could suppress endotoxininduced TNF-α mRNA and protein elevation and inhibit NFkappa B activation in the intestine. However the least dosage of ketamine to inhibit IL-6 was 5 mg/kg in our experiment. CONCLUSION: Ketamine can suppress endotoxin-induced production of proinflammatory cytokines such as TNF-α and IL-6 production in the intestine. This suppressive effect may act through inhibiting NF-kappa B.     

脂多糖和抗炎药物对肺血管内巨噬细胞核因子kB的影响

观察脂多糖致肺血管内巨噬细胞核因子ｋｂ的活化及抗炎药物地塞米松和阿斯匹林对ＮＦ－ｋＢ的影响。方法用改良法分离,培养猪ＰＩＭ,设正常对照,ＬＰＳ刺激,ＤＥＸ和ＡＳＡ干预组,共４组。用凝胶电泳迁移率改变分析法和放射免疫分析法,分别检测ＰＩＭ核提取物ＮＦ－ｋＢ的活性和细胞培养上清肿瘤坏死因子α的含量。     

Differential role of I kappa B kinase 1 and 2 in primary rat hepatocytes

Nuclear factor-kappa B (NF-kappa B) protects hepatocytes from undergoing apoptosis during embryonic development and during liver regeneration. Activation of NF-kappa B is mediated through phosphorylation of its inhibitor, I kappa B, by a kinase complex that contains 2 I kappa B kinases. We analyzed the differential role of I kappa B kinase 1 (IKK1) and I kappa B kinase 2 (IKK2) in tumor necrosis factor alpha (TNF-alpha)- and interleukin-1 beta (IL-1 beta)-mediated NF-kappa B activation in primary rat hepatocytes. Maximal induction of IKK activity was observed 5 minutes after TNF-alpha and 15 minutes after IL-1 beta treatment, and activated IKK was able to phosphorylate GST-I kappa B (1-54) and GST-p65 (354-551), but not a GST-p65 (354-551) substrate with a serine-to-alanine substitution at position 536. Infection with an adenovirus containing catalytically inactive IKK2K44M (Ad5IKK2dn) completely blocked both TNF-alpha- and IL-1 beta-induced GST-I kappa B and GST-p65 phosphorylation, I kappa B degradation, and NF-kappa B DNA binding. Adenovirally transduced, catalytically inactive IKK1K44M (Ad5IKK1dn) reduced IKK activity and NF-kappa B DNA binding only slightly. Accordingly, Ad5IKK2dn induced apoptosis in 75% (+/-6%) of hepatocytes after 12 hours of TNF-alpha, which was accompanied by activation of caspases 3 and 8, nuclear fragmentation, and DNA laddering. In contrast, Ad5IKK1dn led to 21% (+/-2%) apoptosis in TNF-alpha-treated hepatocytes after 12 hours and comparatively low activity of caspases 3 and 8. Furthermore, Ad5IKK2dn completely blocked the induction of inducible nitric oxide synthase (iNOS), whereas Ad5IKK1dn had no influence on the expression of iNOS. Thus, IKK2 is the main mediator for cytokine-induced NF-kappa B activation in primary hepatocytes and protects against TNF-alpha-induced apoptosis, whereas IKK1 kinase activity is not required for NF-kappa B activation.     

Toll样受体参与小鼠肝脏缺血再灌注损伤

目的探讨Toll样受体是否参与小鼠肝脏缺血再灌注损伤及其机制. 方法用Toll样受体缺损小鼠(C3H/Hej,Hej组)和野生型(C3H/Heouj,Heouj组)小鼠复制部分肝脏缺血再灌注损伤模型,于缺血45min,再灌注1h和3h处死动物,检测血清天门冬氨酸氨基转移酶(AST)和血清肿瘤坏死因子α(TNFα)的含量;并以northern blot及髓过氧化物酶(MPO)试验分别检测缺血肝组织TNFα mRNA的表达和MPO的含量. 结果 (1)再灌注1、3h,与假手术组相比,小鼠血浆AST明显升高,但Hej组明显低于Heouj组(661.83U/L±106.09U/L和1215.5U/L±174.03U/L,t＝-6.65,P<0.01;1145.17U/L±132.43U/L和2958.17U/L±186.81U/L,t＝-5.57,P<0.01);(2)再灌注3h时,与假手术组相比,Hej组和Heouj组小鼠血清TNFα浓度明显升高,且前者明显低于后者(152.39pg/ml±43.3pg/ml和249.12pg/ml±51.89pg/ml,t＝-3.13,P<0.05);(3)再灌注1h,除假手术组外,Hej组和Heouj组小鼠缺血肝组织内可见TNFα mRNA的表达,但前者的表达水平明显低于后者,杂交带密度分析显示两者之间差异有显著性 (80.3±28.8与189.4±24.6,t＝-3.25,P<0.05);(4)再灌注3h,与假手术组相比,Hej组和Heouj组小鼠缺血肝组织内MPO含量明显升高,且前者含量明显低于后者(0.059±0.004和0.173±0.025,F＝33.49,P<0.01). 结论 Toll样受体可能通过其介导的炎性通路参与了小鼠肝脏缺血再灌注损伤.     

4-羟基壬烯醛引起支气管上皮细胞白细胞介素-8升高的机制及银杏内酯B的拮抗作用

目的 探讨4-羟基壬烯醛(4-HNE)引起人支气管上皮细胞(16HBE)前炎因子白细胞介素-8(IL-8)升高的机制,以及银杏内酯B对其的拮抗作用.方法 实验分组为4-HNE(10 μmol/L)组、银杏内酯B(100 μmoL/L)孵育+4-HNE(10μmoL/L)组和正常对照组3组,在刺激支气管上皮细胞0.5、2、4、8、12 h后,检测细胞IL-8和IL-8 mRNA的表达水平、磷酸化c-Jun氨基末端激酶(JNK)、p38丝裂原活化蛋白激酶(p38MAPK)、细胞外信号调节蛋白激酶(ERK)1/2、转录激活蛋白-1(AP-1)的活性;观察MAP激酶(MEK)1抑制剂PD98059阻断ERK1信号转导途径后,对4-HNE引起的AP-1结合活性和IL-8生成的影响.检测上述3个实验组AP-1的结合活性.应用方差分析和t检验进行统计学处理.结果 4-HNE组、银杏内酯B孵育+4-HNE组、正常对照组刺激细胞4 h后,细胞上清IL-8值分别为(98.3 ±4.2)、(88.2±5.3)和(65.3±6.2)μg/L;刺激12 h后细胞上清IL-8值分别为(116.5±5.6)、(102.8±4.7)和(63.7±6.6)μg/L.4-HNE组0.5、2、4、8、12 h的磷酸化ERK1水平高于对照组(t值为2.83～14.03,P均<0.05);4-HNE、银杏内酯B孵育+4-HNE组、PD98059+4-HNE组及正常对照组的AP-1结合活性差异有统计学意义(F值为21.49～194.16,P均<0.01).PD98059孵育2 h,4-HNE作用2、4、8、12 h后,IL-8表达和AP-1结合活性明显低于未用PD98059孵育4-HNE刺激组.银杏内酯B+4-HNE组AP-1结合活性低于4-HNE组.结论 4-HNE可以通过ERK1途径增强AP-1转录活性,促进支气管上皮细胞IL-8表达.而银杏内酯B可通过抑制AP-1活性,减少4-HNE引起的支气管上皮细胞IL-8的合成.     

红霉素对肺纤维化大鼠核因子κB活性及细胞因子mRNA表达 …

目的 探讨红霉素治疗肺纤维化的机制和疗效。方法 实验用Ｗｉｓｔａｒ大鼠８１只,分成三组;正常一附属博莱霉素模型组和红霉素治疗组。每组２７只。气管内注入单剂量博莱霉素制备大鼠肺纤维化模型,于博莱霉素气管内注入后每日给予口红霉素治疗,各组动物分别于气管内用药后第４,７和２８天处死动物。用凝胶电泳迁移实验测定肺泡巨噬细胞的核因子（ＮＦ）－κＢ活性;     

结核分支杆菌抗原85B DNA疫苗转染人气道上皮细胞对哮喘患者Th1/Th2平衡的调节作用

目的　研究结核分支杆菌抗原 85B(简称Ag85B)DNA疫苗经人气道上皮转化后的培养上清液对尘螨过敏哮喘患者外周血单个核细胞 (PBMC)Th1/Th2细胞因子释放的调控作用。方法亚克隆构建表达Ag85B基因的真核表达载体pMG Ag85B ,经脂质体介导转化离体培养气道上皮细胞16HBE ,并收集转化细胞培养上清液 ,用逆转录 聚合酶链反应 (RT PCR)方法检测转基因气道上皮Ag85BmRNA水平的表达 ,螨过敏哮喘组 (18例 )和正常组 (13例 )PBMC在离体单独与螨或与Ag85B转染上清液共同培养 ,酶联免疫吸附测定 (ELISA)法测定其培养上清液中白介素 5 (IL 5 )、γ干扰素 (IFN γ)的水平。结果　RT PCR法扩增转染的 16HBE细胞中有 992bp长度的预期片断。无刺激状态下 ,哮喘患者和正常人PBMC培养上清液的IL 5、IFN γ无差异 ,螨刺激可促进哮喘患者PBMC分泌IL 5 ,刺激和未刺激处理分别为 (17 1± 1 9)pg/ml和 (11 9± 0 9)pg/ml(P =0 0 2 ) ;螨 +Ag85B转染上清液与单纯螨刺激比较 ,前者IFN γ水平显著高于后者 ,分别为 (111 5± 15 9)pg/ml和 (5 8 8± 7 8)pg/ml(P =0 0 4 ) ,IFN γ/IL 5的比值分别为 7 2± 4 8和 4 9± 2 5 (P =0 0 8)。正常人不同刺激方法间比较 ,所有指标差异均无显著性 (P >0 0 5 )。结论　螨过敏的变应性     

Amnion-derived cells express intercellular adhesion molecule-1: regulation by cytokines

We have examined the expression of the intercellular adhesion molecule-1 (ICAM-1) mRNA in primary and established amnion-derived cell cultures and regulation of this expression by tumour necrosis factor-alpha (TNF-alpha) and interleukin (IL)-1beta. TNF-alpha (50 ng/ml) and IL-1beta (1.0 ng/ml) induced 18- and 11-fold increases respectively in expression of the ICAM-1 mRNA in WISH cells (an amnion epithelium-derived cell line). The increase was detectable within one hour of treatment and peaked by two hours. The protein synthesis inhibitor, cycloheximide (10 microg/ml) did not inhibit this induction. Increased levels of ICAM-1 protein were detected in the cells within 4 h after initiation of treatment with either cytokine. By 16 h of treatment with IL-1beta or TNF-alpha ICAM-1 reached 40 and 73 pg/microg cellular protein, representing 6- and 11-fold stimulations respectively. In primary amnion cells, basal expression of ICAM-1 mRNA was undetectable. However, TNF-alpha (50 ng/ml) induced ICAM-1 mRNA within two hours, peak expression being reached between four and eight hours after initiation of treatment. The present report demonstrates for the first time that amnion derived cells can express ICAM-1 and, further, that this expression is regulated by pro-inflammatory cytokines. This has implications for the amnion as a possible source for soluble ICAM-1, for this gene product as a marker for preterm labour, and for participation of the amnion, additional to its reported secretory role, in inflammatory processes of the fetal membranes.     

Production and modulation of interleukin 6 synthesis by synoviocytes derived from patients with arthritic disease.

Interleukin 6 (IL-6) is a potent cytokine, the biological activities of which include the stimulation of immunoglobulin secretion, T cell activation, induction of the acute phase response, activation of megakaryocytes, and pyrogenicity. These biological activities make it a plausible contributor to rheumatoid arthritis. The ability of synoviocytes to synthesise this potential mediator of inflammation was tested. Cultures of fibroblast-like cells were established from joint tissue from patients with rheumatoid arthritis, degenerative joint disease, or trauma. Supernatants from synoviocytes from each diagnostic category contained IL-6-like activity as detected in a B9 plasmacytoma cell proliferation assay. Supernatants from IL-1 stimulated synoviocytes from patients with rheumatoid arthritis (n = 5) contained an average of 70,000 U/ml IL-6. Western blot analysis confirmed that these supernatants contained peptides that reacted with a highly specific antibody to IL-6. A cDNA probe specific for IL-6 hybridised with mRNA derived from synoviocytes representative of each disease state. Interleukin 6 mRNA expression increased by culturing synoviocytes in the presence of 10% calf serum, IL-1 (30 U/ml), insulin (166 ng/ml), or basic fibroblast growth factor (16 ng/ml). In contrast, dexamethasone (10(-6) mol/l) suppressed the ability of IL-1 to increase the expression of IL-6 mRNA. Recombinant IL-6 itself did not detectably upregulate its own message. The regulation of production of IL-6 by synoviocytes may be important in the pathogenesis of joint inflammation.