慢性阻塞性肺疾病患者痰中肥大细胞和嗜酸粒细胞因子变化及其相关性研究

目的探讨慢性阻塞性肺疾病(COPD)患者类糜蛋白酶(chymase)活性,类胰蛋白酶(tryptase)、白细胞介素8(IL8)、嗜酸粒细胞趋化因子(eotaxin)水平和中性粒细胞(NEU)、嗜酸粒细胞(EOS)计数的相关性及临床意义。方法老年COPD患者73例(重度21例、中度21例、轻度31例),采用双抗体夹心酶联免疫吸附测定,检测诱导痰IL8、eotaxin水平。在UniCAP100全自动体外变应原检测仪上进行tryptase检测,Chymase活性测定使用琥珀酰丙氨酸丙氨酸脯氨酸苯丙氨酸酰苯氨(SAAPP)作为底物,采用酶标仪在410nm连续监测吸光度的变化。结果(1)老年急性加重期COPD患者(重、中、轻)痰tryptase的中位数分别为2840、2150、595ng/L,治疗后各组痰tryptase水平显著下降(分别为1510、920、33ng/L,P均<001)。加重期重度、中度与轻度患者比较差异均有显著性(P均=0)。急性加重期痰IL8、痰eotaxin中位数分别为12998、4549、787ng/L;227、151、74ng/L。重度、中度组痰IL8、eotaxin水平均高于轻度组(P均<001)。治疗后痰IL8、eotaxin中位数分别为10375、3266、679ng/L;79、63、68ng/L。重度、中度患者痰IL8、eotaxin水平均比治疗前显著降低(P均<001)。(2)重、中度COPD患者急性加重期痰chymase活性高于轻度组,黄豆胰蛋白酶抑制剂(SBTI)、α1抗胰蛋白酶(α1A     

嗜酸性粒细胞趋化因子在变应性鼻炎体质模型鼻腔黏膜中的表达及意义

目的:研究不同体质模型中变应性鼻炎的鼻腔黏膜嗜酸性粒细胞趋化因子(Eotaxin)的表达及意义。方法:采用中药灌饲及同类传代的方法构建不同的体质模型;在此基础上采用卵清蛋白腹腔注射及滴鼻的方法构建变应性鼻炎的疾病模型,瑞氏染色计数鼻黏膜组织的嗜酸性粒细胞和肥大细胞,免疫组化检测鼻腔黏膜Eotaxin的表达。结果:寒、热、常体的变应性鼻炎体质模型成功建立,鼻腔黏膜Eotaxin在各组体质的变应性鼻炎中均能表达,且寒体组表达最明显(P0.05),并与鼻黏膜组织中的嗜酸性粒细胞和肥大细胞呈正相关(P0.05)或显著正相关(P0.01)。结论:体质在变应性鼻炎的免疫调节中起关键作用,能够影响Eotaxin的表达,而Eotaxin募集、趋化了嗜酸性粒细胞和肥大细胞在鼻腔黏膜的浸润和表达,能够反映变应性鼻炎的严重程度。     

Eotaxin原核表达载体的构建及初步表达

目的：获得人嗜酸性粒细胞趋化因子Eotaxin基因，构建其原核表达载体，并初步表达成熟的Eotaxin，为进一步构建其N-端突变体，检测其生物学活性作准备。方法：提取外周血单个核细胞细胞总RNA，经RT-PCR扩增编码人Eotaxin全长cDNA序列，并将其克隆至T载体进行序列测定，然后构建于原核表达载体pET30a^＋中，并转入大肠杆菌表达。结果：RT-PCR扩增得到了400bp左右的DNA片段，序列分析发现其中包含了GenBank中报道的编码EotaxincDNA序列，并获得了正确表达。结论：EotaxincDNA的克隆及其原核表达载体的构建及表达，为进一步构建其N-端突变体，纯化和检测其生物学活性奠定了基础。     

Differential release of MIP-1α and eotaxin during infection of mice by <Emphasis Type="Italic">Histoplasma capsulatum</Emphasis> or inoculation of β-glucan

Objective:In the present study, we evaluated the levels of MIP-1 and eotaxin and in vivo migration in the peritoneal cavity model, in mice inoculated with live yeast forms of Histoplasma capsulatum or the -glucan cell wall component of this fungus, and the influence of a leukotriene biosynthesis inhibitor, MK886, on the release of these chemokines in relation to cell recruitment. Materials:Female outbred Swiss mice (N = 4–5 per group, 3–4 wk, were used. Mice were injected i.p. with 1 ml of the 6×10⁵ live yeast form of the fungus or with 10 g of -glucan from the cell wall fraction, and treated daily with MK886 (1 mg kg^-1, p.o.) or vehicle. Results:The fungus induced rapid generation of high levels of MIP-1, which remained elevated from 4–48 h whereas very little eotaxin was detected at any time point (Fig. 1A and B). In contrast, the -glucan induced a little MIP-1 but considerably higher concentrations of eotaxin within the first four hours; however, the level of neither chemokine was sustained (Fig. 2A and B). Treatment of animals with MK886 was effective in reducing the numbers of neutrophils, eosinophils and, to a lesser degree, mononuclear cells accumulating in the peritoneal cavity in response to both the live fungus (Fig. 1C–E) and the cell wall -glucan (Fig. 2C–E). Conclusions:The results suggest that chemokines and leukotrienes may play key roles in the inflammatory cell influx to H. capsulatum infection or to the inoculation of the -glucan cell wall component of this fungusReceived 28 May 2003; returned for revision 16 January 2004; accepted by I. Ahnfelt-Rønne 7 March 2004     

雷公藤甲素对哮喘小鼠信号转导和转录激活因子6及eotaxin表达的影响

目的研究雷公藤甲素对哮喘小鼠信号转导和转录激活因子6(STAT6)、嗜酸粒细胞趋化因子(eotaxin)表达的影响,探讨其治疗哮喘的机制。方法建立小鼠卵蛋白哮喘模型,随机分为7组,分别为对照组、地塞米松治疗组及不同剂量雷公藤甲素治疗组,测定支气管肺泡灌洗液(BALF)中白细胞总数及嗜酸粒细胞(Eos)计数;逆转录-聚合酶链反应(RT-PCR)及免疫组织化学法分别检测肺组织中STAT6、eotaxin mRNA及气道上皮STAT6、eotaxin蛋白表达水平。结果哮喘组STAT6、eotaxin mRNA及蛋白表达明显高于正常对照组(P<0.01),雷公藤甲素治疗组及地塞米松组明显低于哮喘组(P<0.01)。气道上皮STAT6的蛋白表达与BALF中白细胞数、Eos计数及气道上皮eotaxin蛋白表达呈正相关(r分别为0.528,0.612,0.682,P<0.01)。气道上皮eotaxin蛋白表达与白细胞总数、Eos计数呈正相关(r分别为0.79,0.88,P<0.01)。结论雷公藤甲素抑制哮喘气道炎症的机制可能与其抑制STAT6及eotaxin的表达活性有关。     

患儿血清嗜酸细胞趋化因子、可溶性细胞间粘附分子-1的检测

目的 探讨哮喘患儿血清嗜酸细胞趋化因子(Eotaxin)、可溶性细胞间粘附分子-1(SIcAm-1)水平相关性及临床意义.方法 哮喘患儿30例,采用双抗体夹心酶联免疫吸附试验(SELISA)测定其血清Eotaxin、SIcAm-1质量浓度.结果 哮喘患儿急性期血清中Eotaxin高于对照组(P＜0.01)及间歇组(P＜0.01),间歇期也高于对照组(P＜0.05).哮喘患儿急性期血清中SIcAm-1高于对照组(P＜0.01)及间歇组(P＜0.01),间歇期也高于对照组(P＜0.05).Eotaxin、SIcAm-1血清水平存在相关性(P＜0.01).结论 Eotaxin、SIcAm-1参与小儿哮喘病理过程,可作为哮喘病气道炎症诊断的重要指标.     

截喘汤对哮喘模型小鼠外周血浆IL-5、TNF-α及Eotaxin的影响

目的:研究中药截喘汤对哮喘模型小鼠外周血浆白细胞介素-5(IL-5)、肿瘤坏死因子(TNF-α)及嗜酸细胞活化趋化因子(Eotaxin)的影响,探讨其治疗支气管哮喘的作用机制。方法:将84只小鼠随机均分为7组,即正常对照组、哮喘模型组、地塞米松组、联合用药组及中药低、中、高剂量组,通过中药血清药理学,体内观察不同剂量截喘汤对哮喘模型小鼠外周血IL-5、TNF-α及Eotaxin含量的影响。结果:截喘汤能通过抑制IL-5和TNF-α的合成而阴滞气道变应性炎症(AAI)产生、发展和持续的各个环节,抑制气道高反应性(BHR),并可通过抑制Eotaxin的表达而减少嗜酸性粒细胞(EOS)在肺部的聚集和活化,从而减轻气道EOS浸润性AAI及BHR。结论:截喘汤可为临床用药提供理论依据。     

Galectin-3对哮喘豚鼠模型炎性细胞因子和趋化因子的干预作用

目的　观察哮喘豚鼠模型中白细胞介素- 5 (IL - 5 )、Eotaxin和C- C趋化因子受体3(CCR- 3)在嗜酸粒细胞炎症中的表达和Galectin- 3的干预作用。方法　32只豚鼠随机分为正常对照组、哮喘组、Galectin- 3干预组和地塞米松(DXM)干预组。卵蛋白(OVA)致敏和激发制作哮喘模型,Galectin- 3干预组和DXM干预组分别于致敏后第14～16 d激发前1h腹腔注射Galectin- 3(0 .5 m g/ kg)和DXM(1mg/ kg) ,每日1次。观察血、骨髓涂片,肺组织切片细胞总数和嗜酸粒细胞(EOS)比例。肺组织切片用IL - 5、CCR- 3及Eotaxin多克隆抗体染色,光镜下分别计数阳性细胞比例。结果　与正常对照组相比较,哮喘组豚鼠外周血、骨髓、肺组织中EOS计数显著增加(P<0 .0 5 ) ,肺组织中IL- 5、CCR- 3和Eotaxin阳性细胞比例显著升高(P<0 .0 5 ) ,DXM干预后EOS计数、IL- 5、CCR- 3和Eotaxin阳性细胞比例显著降低(P<0 .0 5 ) ,Galectin- 3干预对EOS计数和IL - 5阳性细胞比例的影响与DXM组相当(P>0 .0 5 ) ;Galectin- 3也可下调CCR- 3阳性细胞的表达(P<0 .0 5 ) ,作用弱于DXM(P<0 .0 5 ) ;但Galectin- 3对Eotaxin水平无显著影响(P>0 .0 5 )。结论　哮喘动物模型肺内IL - 5、Eotaxin及其受体CCR- 3表达增强。Galectin- 3可抑制IL -5表达,轻度下调CCR- 3     

沙美特罗对哮喘模型大鼠气道炎症的作用及其机制

目的:观察沙美特罗(SLM)对哮喘模型大鼠支气管肺泡灌洗液(BALF)中炎性因子的影响,探讨其在哮喘治疗中的作用机制.方法:选取SD大鼠48只随机分为空白对照组、模型组、SLM组及普萘洛尔加SLM组.收集各组大鼠BALF,光镜下计数嗜酸性粒细胞(EOS),原位末端标记法(TUNEL)检测EOS凋亡指数,酶联免疫吸附试验...     

Reduced ccl11/eotaxin mediates the beneficial effects of environmental stimulation on the aged hippocampus

A deterioration in cognitive performance accompanies brain aging, even in the absence of neurodegenerative pathologies. However, the rate of cognitive decline can be slowed down by enhanced cognitive and sensorimotor stimulation protocols, such as environmental enrichment (EE). Understanding how EE exerts its beneficial effects on the aged brain pathophysiology can help in identifying new therapeutic targets. In this regard, the inflammatory chemokine ccl11/eotaxin-1 is a marker of aging with a strong relevance for neurodegenerative processes. Here, we demonstrate that EE in both elderly humans and aged mice decreases circulating levels of ccl11. Interfering, in mice, with the ccl11 decrease induced by EE ablated the beneficial effects on long-term memory retention, hippocampal neurogenesis, activation of local microglia and of ribosomal protein S6. On the other hand, treatment of standard-reared aged mice with an anti-ccl11 antibody resulted in EE-like improvements in spatial memory, hippocampal neurogenesis, and microglial activation. Taken together, our findings point to a decrease in circulating ccl11 concentration as a key mediator of the enhanced hippocampal function resulting from exposure to EE.