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核因子-κB在蛋白激酶C对支气管哮喘T淋巴细胞增殖和凋亡调控信号转导中的作用 总被引:2,自引:0,他引:2
目的 探讨核因子 κB(NF κB)在蛋白激酶C(PKC)对支气管哮喘T淋巴细胞增殖和凋亡调控的信号转导中的作用。方法 哮喘组和正常对照组的豚鼠以及急性发作期哮喘患者和正常对照者的外周血中分离出T淋巴细胞并分别加入PKC激动剂 12 肉豆蔻酰 13 乙酸佛波酯 (PMA)和NF κB抑制剂二硫代氨基甲酸吡咯烷 (PDTC)培养。用免疫组化法检测NF κB的表达 ,用四甲基偶氮唑盐微量比色法测定增殖反应 ,用原位末端终止法观测凋亡。结果 加入PMA培养的哮喘T淋巴细胞NF κB活化细胞百分比和增殖率与空白对照及加入PMA培养的正常T淋巴细胞相比均显著升高 (P <0 .0 1) ,而加入PDTC后以上指标均显著降低 (P <0 .0 1)。加入PMA培养的哮喘T淋巴细胞的凋亡指数与空白对照及加入PMA培养的正常T淋巴细胞相比均显著降低 (P <0 .0 1) ,加入PDTC后以上指标均显著升高 (P <0 .0 1)。T淋巴细胞NF κB活化细胞的百分比与增殖率均呈显著正相关 (r =0 .5 1~ 0 .72 ,P <0 .0 0 1) ,与凋亡指数均呈显著负相关 (r= 0 .5 5~ 0 .71,P <0 .0 0 1)。结论 T淋巴细胞PKC活化后导致增殖增加及凋亡减少的生物信号可能是通过激活NF κB来转导的。T淋巴细胞PKC NF κB信号转导途径的激活可能是哮喘的发病机制之一 相似文献
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目的 研究重组白细胞介素18(rIL-18)对肺炎链球菌肺炎小鼠Th1/ Th2免疫应答的影响.方法 鼻腔接种肺炎链球菌建立小鼠肺炎链球菌肺炎模型,将Balb/c小鼠24只随机分为3组,分别为对照组,肺炎组和肺炎rIL-18干预组(n=8 ),RT-PCR法检测各组小鼠肺组织中IFN-γ、IL-4 mRNA 的表达,同时支气管肺泡灌洗液(BALB)进行活菌计数,有核细胞分类计数.结果 ①肺炎rIL-18干预组BA LF中性粒细胞和巨噬细胞计数显著高于肺炎组和对照组(P<0.001);②肺炎rIL-18 干预组BALF活菌计数显著低于肺炎组(P<0.001);③肺炎rIL-18干预组肺组织IFN- γ mRNA表达上调而IL-4 mRNA表达下调(P<0.001).结论 在小鼠肺炎链球菌肺炎早期给予rIL-18可诱导IFN-γ的合成,促进Th1免疫应答,使Th1/ Th2免疫平衡向Th1免疫偏移、促进宿主对肺炎链球菌的防御. 相似文献
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磷脂酰肌醇3激酶对TNF-α诱导的大鼠气道平滑肌细胞增殖、凋亡和转化生长因子β1表达的影响 总被引:1,自引:0,他引:1
目的 探讨磷脂酰肌醇3激酶(PI3-K)对TNF-α诱导的大鼠气道平滑肌细胞(ASMC)增殖、凋亡和转化生长因子β1(TGF-β1)表达的影响.方法 以肿瘤坏死因子-α(TNF-α)及PI3-K特异性抑制剂wortmannin作为工具药,分别将TNF-α、TNF-α+wortmannin接种ASMC,置37℃,5%CO2培养箱中培养.逆转录聚合酶链式反应(RT-PCR)检测PI3-Kp85α、TGF-β1 mRNA表达,免疫细胞化学染色法检测PI3-Kp85α、PCNA、Bcl-2蛋白表达及定位,Sandwich ELISA检测NF-κB活性,四甲基偶氮唑蓝(MTT)微量比色法测定ASMC增殖,AnnexinV/PI双标记流式细胞仪分析法检测细胞凋亡,ELISA测定TGF-β1蛋白质含量.结果 (1)培养的ASMC PI3-K p85α的阳性定位在胞浆.TNF-α组ASMC PI3-Kp85α mRNA及蛋白表达水平均显著高于对照组及TNF-α+wortmannin组(P<0.01);(2)TNF-α组ASMCNF-κB活性与对照组相比显著增高(P<0.01),TNF-α+wortmannin组NF-κB活性与TNF-α组相比显著降低(P<0.01);(3)TNF-α组ASMC的增殖反应与对照组及TNF-α+wortmannin组相比显著增加(P<0.01);TNF-α组细胞凋亡率与对照组相比差异无统计学意义(P>0.05),TNF-α+wortmannin组细胞凋亡率显著高于TNF-α组及对照组(P<0.01).(4)TNF-α组ASMC的PCNA、Bcl-2蛋白表达量均显著高于对照组及TNF-α+wortmannin组(P<0.01);(5)TNF-α组ASMC的TGF-β1的mRNA表达水平、培养液上清TGF-β1的蛋白质含量均显著高于对照组及TNF-α+wortmannin组(P<0.01).结论 PI3-K可能参与调控TNF-α诱导的大鼠气道平滑肌细胞的增殖、凋亡及TGF-β1的表达和分泌,NF-κB作为PI3-K的下游信号参与此过程. 相似文献
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survivin基因在非小细胞肺癌中的表达及与P53、c-myc、k-ras蛋白表达的关系 总被引:43,自引:3,他引:43
目的:探讨survivin基因在非小细胞肺癌(NSCLC)中的表达,及与P53、c-myc、k-ras蛋白表达的相互关系。方法:逆转录PCR法检测了76例NSCLC肿瘤组织,20例良性瘤样病变和21例病灶旁正常肺组织survivin mRNA表达,免疫组织化学法检测P53,c-myc,k-ras蛋白表达,并将结果进行了相关分析。结果:61%NSCLC癌组织表达survivin mRNA基因,而良性瘤样病变和正常组织则分别为30%和19%(P<0.001)。survivin基因表达与肺癌组织细胞类型,分化程度,TNM分期及淋巴结转移无明显相关关系。P53蛋白,c-myc与survivin基因表达显著相关,k-ras蛋白表达与survivin基因表达未见明显相关。结论:(1)survivin基因在肺癌组织中表达上调,提示该基因对NSCLC发生发展起重要作用。Survivin基因有望成为肺癌基因治疗的新靶点。(2)抑癌基因P53的失活和癌基因c-myc的上调与survivin基因的表达可能在NSCLC癌变中起协同作用。survivin和k-ras基因则可能通过各自不同的机制参与NSCLC的发病机制。 相似文献
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In order to explore the expression of PI-3K in T lymphocytes of asthmatic rats and the relationship between PI-3K and activation of T lymphocytes, 24 Wistar rats were randomly divided into 4 groups: normal control group, asthmatic one-week group, asthmatic two-week group and asth-matic four-week group. T cells were purified from blood of each rat and the expression of PI-3K was observed by immunocytochemical fluorescence staining, the semiquantitative fluorescence intensity was measured by HPIAS-2000 analytic software, and the expression of IL-4 in supernatants was de-tected by ELISA. The results showed that the fluorescence intensity of T lymphocytes in asthmatic groups was significantly higher than that in normal control (P<0.001), indicating that the expression of PI-3K in T lymphocytes of asthmatic rats was significantly higher than that in those of normal controls, and the difference between acute and chronic stage asthmatic groups was significant (P<0.05). The expression levels of IL-4 protein in supernatants of asthmatic T lymphocytes were sig-nificantly higher than those in the normal controls (P<0.05). There was a significant positive correla-tion between the expression of PI-3K in T lymphocytes and the IL-4 protein expression in super-natants (r=0.583, P<0.01). It was suggested that PI-3K signal pathway may participate in the proc-esses of activation and other cytological effects of asthmatic T lymphocytes, thus may play an impor-tant roles in the pathogenesis of asthma. 相似文献
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Whether inhibiting the activity of nuclear factor (NF)-κB potentiates cisplatin-induced apoptosis in non-small cell lung cell line A549 cells was investigated. The recombinant plasmid pcDNA3.1(+)/IκBα expressing IκBα was constructed. The in vitro cultured A549 cells were transfected with pcDNA3.1 (+)/IκBα alone, or pcDNA3.1(+)/IκBα combined with cisplatin. The mitochondrial membrane potential (△ψm) was determined by rhodamine 123, the activity of caspase-3 was tested by colorimetric assay, and cell apoptosis was detected by flow cytometry with the annexin V/propidium iodide assay. The results showed that the activity of NF-κB in A549 cells was inhibited by transfecting pcDNA3.1(+)/IκBα. Transfection of pcDNA3.1(+)/IκBα alone did not promote apoptosis. Treatment of cisplatin alone had a little effect on cell apoptosis. Transfection of pcDNA3.1(+)/IκBα combined with cisplatin treatment significantly induced apoptosis of A549 ceils. It was concluded that inhibiting the activity of NF-κB potentiated cisplatin-induced apoptosis of A549 cells. 相似文献
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COPD急性加重期激素治疗和机械通气的应用 总被引:1,自引:0,他引:1
熊维宁 《实用医学进修杂志》2008,36(1):6-9
慢性阻塞性肺疾病(chronic obstruc—tire pulmonary diseases,COPD)急性加重期(AECOPD)的治疗内容较多,包括确定COPD急性加重的原因、COPD急性加重的诊断和严重性评价、院外治疗和住院治疗等4个大的方面。而住院治疗主要的治疗方案又包括评估病情的严重程度、控制性02疗、抗生素、支气管舒张剂、糖皮质激素(简称激素)、机械通气及其他治疗措施等7个方面。其中激素和机械通气的应用是治疗的热点和难点,现分别述及。 相似文献
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目的 探讨联合检测支气管肺泡灌洗液(BALF)中DPC4和LKB1突变对鉴别肺部X线孤立块影良恶性的价值.方法 选择X线检查有肺部孤立块影的患者49例,其中非小细胞肺癌(NSCLC)患者39例,非癌对照组10例,用聚合酶链式反应-单链构象多态性(PCR-SSCP)法检测BALF中DPC4第8~11外显子、LKB1第1~9外显子的突变情况.结果 39例NSCLC患者中DPC4缺失3例,突变12例,LKB1突变10例,1例NSCLC患者同时发生DPC4和LKB1突变,非癌对照组均未检测出抑癌基因缺失或突变.联合检测DPC4和LKB1诊断NSCLC的灵敏度(61.5%,24/39)较单独检测DPC4或LKB1显著增高(P <0.05).结论 用PCR-SSCP技术检测BALF中DPC4或LKB1基因突变可作为鉴别肺部孤立块影良恶性的辅助指标之一,有助于肺癌的早期诊断和分期,可能具有一定的临床应用价值,联合检测两种基因较单独检测灵敏度更高. 相似文献