臭氧和细颗粒物暴露对大鼠心脏自主神经系统和系统炎症的影响

目的探讨臭氧(O3)和大气细颗粒物(PM2.5)暴露对大鼠心脏自主神经系统和系统炎症的影响以及二者之间是否存在关联。方法 48只Wistar大鼠随机分为8组。PM2.5低、中、高剂量单独暴露组为每只动物分别经气管滴注0.2、0.8和3.2mg的细颗粒物。O3和PM2.5联合暴露低、中、高剂量组为大鼠预先暴露于0.8ppm的臭氧4h,然后再气管滴注PM2.50.2、0.8和3.2mg。O3单独暴露组为只吸入暴露4h臭氧,对照组为气管滴注生理盐水,每周暴露2次,共3周。末次暴露24h后,测定血压,同时记录心电图。测定血清中TNF-α、IL-6和CRP。取右心室进行HE染色病理分析。结果 PM2.5单独暴露和联合暴露组中心率变异性(HRV)指标与对照组比较有显著变化。O3单独暴露仅引起低频成分(LF)显著增加。心率只有在联合暴露组有明显降低。血压在联合暴露组和高剂量PM2.5组有明显上升。TNF-α和IL-6在PM2.5单独暴露和联合暴露组均有上升趋势。CRP在PM2.5单独和联合暴露组均表现出明显的剂量-效应关系。IL-6、TNF-α和CRP与HRV之间显著相关。病理学检查发现,PM2.5暴露组有颗粒物的沉积和心肌炎症。结论 O3可增强由PM2.5暴露引起的大鼠心脏自主神经系统紊乱和炎症反应。     

支原体肺炎患儿血清TNF-α、IL-6、IL-8的检测及其临床意义

目的 检测支原体肺炎(MPP)患儿血清中肿瘤坏死因子-a(TNF-a)、白细胞介素-6(IL-6)、白细胞介素-8(IL-8)水平,探讨其临床意义.方法 应用ELISA双抗体夹心法检测44例MPP患儿(研究组)和36例健康儿童(对照组)血清中TNF-α、IL-6、IL-8水平.结果 研究组MPP患儿急性期血清TNF-α、IL-6、IL-8分别为(29.22±1.26)、(20.18±1.32)、(28.01±1.38)pg/ml,高于对照组的(7.98±2.30)、(6.21±0.72)、(7.82±2.00)pg/ml,差异有统计学意义(P＜0.05);研究组恢复期血清TNF-α、IL-6、IL-8分别为(11.02±1.46)、(8.86±1.02)、(10.54±1.07)pg/ml,显著低于急性期(P＜0.05);虽然高于对照组,差异无统计学意义;12例MPP肺纤维化患儿胸水TNF-α、IL-6、IL-8分别为(49.02±2.20)、(32.28±2.54)、(45.90±2.86)pg/ml,较无肺纤维化者的(28.80±1.92)、(26.06±1.77)、(28.86±2.75) pg/ml,显著增高(P＜0.05),12例MPP肺纤维化血清中TNF-α、IL-6、IL-8分别为(30.18±2.86)、(27.84±2.86)、(28.42±3.20)pg/ml,低于其胸水(P＜0.05);6例存在胸腔积液但无肺纤维化改变的患儿,胸水中TNF-α、IL-6、IL-8水平与血清中差异无统计学意义.结论 TNF-α、IL-6、IL-8在MPP的发生、发展中可能起重要作用,并且与肺部有无纤维化形成有关,故在诊断、治疗、预后方面有重要意义.     

Effect of inhaled ozone on lung histamine in conscious guinea pigs

The effect of short-term ozone (O3) exposure on pulmonary mast cell function was examined. Guinea pigs were continuously exposed to 1.0 ppm O3 for 2, 4, and 8 hr. O3 exposure produced a significant decrease in lung histamine concentration. Two-hour exposure to O3 caused a 22.4 +/- 7.0% decrease in lung histamine concentration compared with controls. Ozone exposures of 4 and 8 hr caused lung histamine concentrations to decrease by 43.7 +/- 7.7 and 49.0 +/- 7.5% (P less than 0.05), respectively, without significant changes in lung water or protein, or evidence of cytotoxicity. These results suggest that O3 or its metabolites affect pulmonary mast cell function by stimulating the release of histamine from the lung.     

The effect of ozone exposure on the ability of human surfactant protein a variants to stimulate cytokine production.

Ozone exposure can cause inflammation and impaired lung function. Human surfactant protein A (SP-A) may play a role in inflammation by modulating cytokine production by macrophages. SP-A is encoded by two genes, SP-A1 and SP-A2, and several allelic variants have been characterized for each gene. These allelic variants differ among themselves in amino acids that may exhibit differential sensitivity to ozone-induced oxidation and this may produce functional differences. We studied the effects of SP-A variants before and after ozone exposure on the production of tumor necrosis factor (TNF)-alpha and interleukin (IL)-8. These are important proinflammatory cytokines and are expressed by the macrophage-like THP-1 cells. Eight variants were expressed in vitro, characterized by gel electrophoresis, and studied. These included six single-gene SP-A alleles and two SP-A variants derived from both genes. Variants were exposed to ozone at 1 ppm for 4 hr at 37 degrees C, and we compared their ability to stimulate cytokine (TNF-alpha and IL-8) production by THP-1 cells to air-exposed and unexposed SP-A variants. We found that a) SP-A2 variants (1A, 1A(0), 1A(1) stimulate significantly more TNF-alpha and IL-8 production than SP-A1 variants (6A, 6A(2), 6A(4); b) coexpressed SP-A variants (1A(0)/6A(2), 1A(1)/6A(4) have significantly higher activity than single gene products; c) after ozone exposure, all SP-A variants showed a decreased ability to stimulate TNF-alpha and IL-8 production, and the level of the decrease varied among SP-A variants (26-48%); and d) human SP-A from patients with alveolar proteinosis exhibited a minimal decrease (18% and 12%, respectively) in its ability to stimulate TNF-alpha and IL-8 after in vitro ozone exposure. We conclude that biochemical and functional differences exist among SP-A variants, that ozone exposure modulates the ability of SP-A variants to stimulate cytokines by THP-1 cells, and that SP-As from bronchoalveolar lavage (BAL) fluid of certain alveolar proteinosis patients may be oxidized in vivo.     

Effects of ozone on the cholinergic secretory responsiveness of ferret tracheal glands

Oxidant air pollutants exacerbate several pulmonary diseases. Inhalation of ozone has been shown to induce airway smooth muscle hyperresponsiveness. Oxidant injury could also affect airway secretory mechanisms. We postulated that oxidant exposure would alter the glycoconjugate secretory function of airway submucosal glands. To test this hypothesis we examined the effects of in vivo ozone exposure on the in vitro secretory responsiveness of ferret tracheal glands. Ferrets were exposed to 1 ppm ozone, 24 hr/day for 3 or 7 days. Following exposure, glandular explants, denuded of surface epithelial cells, were prepared and incubated in medium containing 3H-glucosamine for 18 hr. Basal secretion of labeled glycoconjugates was significantly increased 31% following 3 days of ozone exposure (P less than or equal to 0.05) and remained elevated 11% after 7 days of exposure compared to the air-exposed group. After 3 or 7 days of exposure to ozone, tracheal gland responsiveness to carbachol was increased as indicated by significantly lower EC50 values (log molar concentration) of -6.43 +/- 0.04 (n = 6) and -6.50 +/- 0.11 (n = 5), respectively; compared to -6.20 +/- 0.08 (n = 6) for the air-exposed group. There was no difference in carbachol EC50 values for air and 7-day ozone-exposed animals treated with dexamethasone. Dexamethasone did not attenuate the ozone-induced increase in basal secretion. Tracheal gland responsiveness to alpha- or beta-adrenergic agonists was not changed by oxidant exposure. These experiments suggest that oxidant injury not only increases basal secretion of respiratory glycoconjugates but also increases tracheal gland sensitivity to a cholinergic agonist.     

Exposure to concentrated ambient particles does not affect vascular function in patients with coronary heart disease

BACKGROUND: Exposure to fine particulate air pollution is associated with increased cardiovascular morbidity and mortality. We previously demonstrated that exposure to dilute diesel exhaust causes vascular dysfunction in humans. OBJECTIVES: We conducted a study to determine whether exposure to ambient particulate matter causes vascular dysfunction. METHODS: Twelve male patients with stable coronary heart disease and 12 age-matched volunteers were exposed to concentrated ambient fine and ultrafine particles (CAPs) or filtered air for 2 hr using a randomized, double-blind cross-over study design. We measured peripheral vascular vasomotor and fibrinolytic function, and inflammatory variables-including circulating leukocytes, serum C-reactive protein, and exhaled breath 8-isoprostane and nitrotyrosine-6-8 hr after both exposures. RESULTS: Particulate concentrations (mean +/- SE) in the exposure chamber (190+/-37 microg/m(3)) were higher than ambient levels (31+/-8 microg/m(3)) and levels in filtered air (0.5+/-0.4 microg/m(3); p<0.001). Chemical analysis of CAPs identified low levels of elemental carbon. Exhaled breath 8-isoprostane concentrations increased after exposure to CAPs (16.9+/-8.5 vs. 4.9+/-1.2 pg/mL, p<0.05), but markers of systemic inflammation were largely unchanged. Although there was a dose-dependent increase in blood flow and plasma tissue plasminogen activator release (p<0.001 for all), CAPs exposure had no effect on vascular function in either group. CONCLUSIONS: Despite achieving marked increases in particulate matter, exposure to CAPs--low in combustion-derived particles--did not affect vasomotor or fibrinolytic function in either middle-aged healthy volunteers or patients with coronary heart disease. These findings contrast with previous exposures to dilute diesel exhaust and highlight the importance of particle composition in determining the vascular effects of particulate matter in humans.     

人免疫缺陷病毒感染者辅助性T淋巴细胞因子变化及机会性感染的临床观察

目的观察人免疫缺陷病毒human immunodeficiency virus,HIV）感染者不同时期及其机会性感染者血清辅助性T淋巴细胞（helper T-Lymphocytes,Th）细胞因子水平的变化规律。方法正常对照组17例。研究组HIV阳性85例（A期17例,B期29例,C期39例）,其中机会性感染31例。用流式细胞仪检测血CD;T和CDs＋T细胞,用酶联免疫吸附技术（enzyme linked immunosorbent assay,ELISA）检测血清白介素-2（interleukin-2,IL-2）、1-干扰素（interferon-1,IFN-γ）、白介素-6（interleukin-6,IL-6）、白介素-10（interleukin-10,IL-10）,数据采用SPSS11．0软件进行统计。结果研究组CD4^＋T细胞（361．85±230．61）10^6／L低于对照组（772．41±161．56）10^6／L（t=6．992,P〈0．01）,IL-2（61．82±63．59）pg／ml低于对照组（111．25±66．14）pg／ml（t=2．907,P〈0．01）,研究组CD8^＋T细胞（713．36±317．59）10^6／L高于对照组（583．24±96．28）10^6／L（t=3．127,P〈0．01）、IL-10（1362．70±869．49）pg／ml高于对照组（818．54±276．22）pg／ml（t=4．704,P〈0．01）和IL-6（1883．14±1058．61）pg／ml高于对照组（1208．52±745．36）pg／ml（t=2．502,P〈0．05）。随着病程进展,IL-2逐渐下降,C期（51．72±62．28）pg／ml和B期（69．02±62．77）pg／m1分别低于对照组,而IL-6、IL-10逐渐上升,C期的IL-6（2040．27±1078．95）pg／ml、IL-10（1472．10±982．03）pg／ml均高于对照组;B期的IL-10（1347．35±780．95）pg／ml高于对照组（818．54±276．22）pg／ml。机会性感染组IL-6（2236．24±1052．42）pg／ml高于无机会性感染组（1680．43±1017．05）pg／ml（t=2．395,P〈0．05）。结论HIV感染者应动态检测血清IL-2、IL-6、IL-10的变化,同时可考虑上调IL-2和下调IL-6、IL-10,调整机体TH1／TI-L2细胞的平衡,以延缓疾病进展。     

Induction of proinflammatory cytokines and C-reactive protein in human macrophage cell line U937 exposed to air pollution particulates

Exposure to particulate matter air pollution causes inflammatory responses and is associated with the progression of atherosclerosis and increased cardiovascular mortality. Macrophages play a key role in atherogenesis by releasing proinflammatory cytokines and forming foam cells in subendothelial lesions. The present study quantified the inflammatory response in a human macrophage cell line (U937) after exposure to an ambient particulate sample from urban dust (UDP) and a diesel exhaust particulate (DEP). The effect of native UDP and DEP was compared with their corresponding organic extracts (OE-UDP/OE-DEP) and stripped particles (sUDP/sDEP) to clarify their respective roles. Exposure to OE-UDP, OE-DEP, UDP, DEP, and 2,3,7,8-tetrachlorodibenzo-p-dioxin led to a greater increase of interleukin (IL)-8, tumor necrosis factor-alpha, and cyclooxygenase-2 mRNA expression than did the stripped particles, whereas sUDP, sDEP, UDP, and DEP led to a greater production of C-reactive protein and IL-6 mRNA. The particles and the organic extract-induced expression of cyclooxygenase-2 and cytochrome P450 (CYP)1a1 was significantly suppressed by co-treatment with an aryl hydrocarbon receptor (AhR) antagonist, indicating that these effects are mainly mediated by the organic components, which can activate the AhR and CYP1a1. In contrast, the induction of C-reactive protein and IL-6 seems to be a particle-related effect that is AhR independent. The inflammatory response induced by particulate matter was associated with a subsequent increase of cholesterol accumulation, a hallmark of foam cells. Together, these data illustrate the interaction between particulate matter and the inflammatory response as well as the formation of cholesterol-accumulating foam cells, which are early markers of cardiovascular disease.     

Oxidative DNA damage and inflammatory responses in cultured human cells and in humans exposed to traffic-related particles

Particulate pollution is a major public health concern because epidemiological studies have demonstrated that exposure to particles is associated with respiratory diseases and lung cancer. Diesel exhaust particles (DEP), which is classified as a human carcinogen (IARC, 2012), are considered a major contributor to traffic-related particulate matter (PM) in urban areas. DEP consists of various compounds, including PAHs and metals which are the principal components that contribute to the toxicity of PM. The present study aimed to investigate effects of PM on induction of oxidative DNA damage and inflammation by using lymphocytes in vitro and in human exposed to PM in the environment. Human lymphoblasts (RPMI 1788) were treated with DEP (SRM 2975) at various concentrations (25–100 μg/ml) to compare the extent of responses with alveolar epithelial cells (A549). ROS generation was determined in each cell cycle phase of DEP-treated cells in order to investigate the influence of the cell cycle stage on induction of oxidative stress. The oxidative DNA damage was determined by measurement of 8-hydroxy-deoxyguanosine (8-OHdG) whereas the inflammatory responses were determined by mRNA expression of interleukin-6 and -8 (IL-6 and IL-8), Clara cell protein (CC16), and lung surfactant protein-A (SP-A). The results showed that RPMI 1788 and A549 cells had a similar pattern of dose-dependent responses to DEP in terms of particle uptake, ROS generation with highest level found in G2/M phase, 8-OHdG formation, and induction of IL-6 and IL-8 expression. The human study was conducted in 51 healthy subjects residing in traffic-congested areas. The effects of exposure to PM_2.5 and particle-bound PAHs and toxic metals on the levels of 8-OHdG in lymphocyte DNA, IL-8 expression in lymphocytes, and serum CC16 were evaluated. 8-OHdG levels correlated with the exposure levels of PM_2.5 (P < 0.01) and PAHs (P < 0.05), but this was not the case with IL-8. Serum CC16 showed significantly negative correlations with B[a]P equivalent (P < 0.05) levels, but positive correlation with Pb (P < 0.05). In conclusion, a similar pattern of the dose-dependent responses to DEP in the lymphoblasts and lung cells suggests that circulating lymphocytes could be used as a surrogate for assessing PM-induced oxidative DNA damage and inflammatory responses in the lung. Human exposure to PM leads to oxidative DNA damage whereas PM-induced inflammation was not conclusive and should be further investigated.     

装饰材料挥发物吸入染毒对大鼠IL-4、IL-5含量及ICAM-1 mRNA表达的影响

[目的]探讨挥发性有机化合物的吸入和呼吸道变态反应之间的关系，为评价装修后室内空气污染提供毒理学依据。[方法]用装饰材料挥发物对雄性SD大鼠静式吸入染毒30d，测定支气管肺泡灌洗液(BALF)中白细胞介素(IL-4、IL-5)含量和肺组织细胞间黏附分子-1(ICAM-1)mRNA表达。[结果]在中浓度染毒组(甲醛浓度为0．81mg／m^3、总挥发性有机化合物(TVOC)浓度为84．88mg／m^3)，BALF中IL-4含量平均为12．95pg／ml，与对照组相比显著升高；在高浓度染毒组(甲醛浓度为1．50mg／m^3、TVOC浓度为299．03mg／m^3)，BALF中IL-4含量平均为13．23pg／ml，IL-5含量平均为23．21pg／ml，均显著高于对照组。中高浓度染毒组大鼠肺组织ICAM-1 mRAN表达显著增强，病理学检查发现肺组织中嗜酸性粒细胞浸润尤为明显。[结论]室内装修可能与呼吸道变态反应性疾病之间有一定联系。     

梅毒患者血清IL-1β、IL-8表达水平与血清反应素滴度的相关性分析

目的 探讨不同临床分期梅毒患者血清白介素-1β(interleukin-1β,IL-1β)和白介素-8(interleukin-8,IL-8)表达水平与梅毒血清反应素滴度的相关性。 方法 收集40例健康体检者血清、120例梅毒患者血清(分为一期梅毒组、二期梅毒组和隐性梅毒组),采用酶联免疫吸附试验检测各组血清中IL-1β、IL-8 水平,比较各组间IL-1β和IL-8表达水平的差异,采用梅毒甲苯胺红不加热血清试验(toluidine red unheated serum test,TRUST)检测梅毒患者血清反应素滴度,根据TRUST滴度将梅毒患者分为高滴度组和低滴度组,比较各组间IL-1β和IL-8表达水平的差异。 结果 各组血清IL-1β、IL-8表达水平差异有统计学意义(P＜0.05),其中二期梅毒组血清IL-1β[15.10(10.63,19.16)]pg/ml和IL-8[108.67(69.96,139.79)]pg/ml明显高于一期梅毒组、隐性梅毒组和正常组;高滴度组IL-8[86.03(62.25,144.94)]pg/ml明显高于低滴度组[57.21(52.16,86.13)]pg/ml,差异有统计学意义(P＜0.05),但高滴度组IL-1β[11.12(7.18,15.96)]pg/ml与低滴度组[8.43(6.93,14.05)]pg/ml比较差异无统计学意义(P＞0.05);二期梅毒患者血清IL-8水平与反应素滴度呈正相关(r=0.625,P＜0.05)。 结论 梅毒患者血清IL-1β和IL-8在一期梅毒、二期梅毒和隐性患者中均呈高水平表达,且IL-1β和血清IL-8表达水平变化与梅毒的病程进展有一定的相关性。     

Alteration of pulmonary immunity to Listeria monocytogenes by diesel exhaust particles (DEPs). II. Effects of DEPs on T-cell-mediated immune responses in rats

Previously, we showed that diesel exhaust particles (DEPs) suppressed pulmonary clearance of Listeria monocytogenes (Listeria) and inhibited the phagocytosis of alveolar macrophages and their response to Listeria in the secretion of interleukin (IL)-1 beta, tumor necrosis factor alpha, and IL-12. In this report we examined the effects of DEPs and/or Listeria on T-cell development and secretion of IL-2, IL-6, and interferon (IFN)-gamma. We exposed Brown Norway rats to clean air or DEPs at 50 or 100 mg/m3 for 4 hr by nose-only inhalation and inoculated with 100,000 Listeria. Lymphocytes in the lung-draining lymph nodes were isolated at 3 and 7 days postexposure, analyzed for CD4+ and CD8+ cells, and measured for cytokine production in response to concanavalin A or heat-killed L. monocytogenes. Listeria infection induced lymphocyte production of IL-6. At 7 days postinfection, lymphocytes from Listeria-infected rats showed significant increases in CD4+ and CD8+ cell counts and the CD8+/CD4+ ratio and exhibited increased production of IFN-gamma and IL-2 receptor expression compared with the noninfected control. These results suggest an immune response that involves the action of IL-6 on T-cell activation, yielding Listeria-specific CD8+ cells. DEP exposure alone enhanced lymphocyte production of both IL-2 and IL-6 but inhibited lymphocyte secretion of IFN-gamma. In rats exposed to 100 mg/m3 DEPs and Listeria, a 10-fold increase occurred in pulmonary bacterial count at 3 days postinfection when compared with the Listeria-only exposure group. The isolated lymphocytes showed a significant increase in the CD4+ and CD8+ cell counts and the CD8+/CD4+ ratio and exhibited increased IL-2 responsiveness and increased capacity in the secretion of IL-2, IL-6, and IFN-gamma. This T-cell immune response was sufficient to allow the Brown Norway rats to clear the bacteria at 7 days postinfection and overcome the down-regulation of the innate immunity by the acute DEP exposure.     

Activation of the stress axis and neurochemical alterations in specific brain areas by concentrated ambient particle exposure with concomitant allergic airway disease

OBJECTIVE: Exposure to ambient particulate matter (PM) has been linked to respiratory diseases in people living in urban communities. The mechanism by which PM produces these diseases is not clear. We hypothesized that PM could act on the brain directly to stimulate the stress axis and predispose individuals to these diseases. The purpose of this study was to test if exposure to PM can affect brain areas involved in the regulation of neuroendocrine functions, especially the stress axis, and to study whether the presence of preexisting allergic airway disease aggravates the stress response. DESIGN: Adult male rats (n = 8/group) with or without ovalbumin (OVA)-induced allergic airway disease were exposed to concentrated air particles containing PM with an aerodynamic diameter pound 2.5 microm (PM(2.5)) for 8 hr, generated from ambient air in an urban Grand Rapids, Michigan, community using a mobile air research laboratory (AirCARE 1). Control animals were exposed to normal air and were treated with saline. MEASUREMENTS: A day after PM(2.5) exposure, animals were sacrificed and the brains were removed, frozen, and sectioned. The paraventricular nucleus (PVN) and other brain nuclei were microdissected, and the concentrations of aminergic neurotransmitters and their metabolites were measured using high-performance liquid chromatography with electrochemical detection. Serum corticosterone levels were measured using radioimmunoassay. RESULTS: A significant increase in the concentration (mean +/- SE, pg/microg protein) of norepinephrine in the PVN was produced by exposure to concentrated ambient particles (CAPs) or OVA alone (12.45 +/- 2.7 and 15.84 +/- 2.8, respectively) or after sensitization with OVA (19.06 +/- 3.8) compared with controls (7.98 +/- 1.3 ; p < 0.05). Serum corticosterone (mean +/- SE, ng/mL) was significantly elevated in the OVA + CAPs group (242.786 +/- 33.315) and in the OVA-presensitized group (242.786 +/- 33.315) compared with CAP exposure alone (114.55 +/- 20.9). Exposure to CAPs (alone or in combination with OVA pretreatment) can activate the stress axis, and this could probably play a role in aggravating allergic airway disease.     

Serum cytokine levels in alcohol-related liver cirrhosis.

Chronic alcoholism complicated by alcoholic liver disease is characterized by activation of the inflammatory response system. To evaluate the role of cytokines in the progress of alcoholic cirrhosis, we assessed serum level of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, and IL-8 and the antiinflammatory cytokines IL-2, IL-10, and transforming growth factor (TGF)-beta in patients with compensated and decompensated alcoholic liver cirrhosis. Compensated alcoholic cirrhosis was characterized by increased IL-6 (6.3+/-2.9 vs. HP 2.2+/-1.4 pg/ml in controls) and decreased IL-10 (HP 4.1+/-3.5 vs. 6.4+/-5.4 pg/ml in controls). TNF-alpha, IL-8, and TGF-beta1 levels were comparable to those found in controls. In sera of patients with decompensated alcoholic liver cirrhosis, besides increased IL-6 (11.2+/-7.7 pg/ml), increased concentrations of TNF-alpha (25.1+/-4.5 vs. 9.1+/-7.0 pg/ml in controls) and IL-8 (171.7+/-294.0 vs. 2.7+/-2.9 pg/ml in controls) were also detected. TGF-beta1 and IL-10 levels were similar to those found in controls. These results strongly indicate that a significant derangement of the balance between proinflammatory and antiinflammatory signals is characteristic of compensated and especially of decompensated alcoholic cirrhosis.     

小儿细菌性肺炎支气管肺泡灌洗液IL-6及IL-8变化的探讨

目的探讨支气管肺泡灌洗液（BALF）中白细胞介素（IL）-6、IL-8与小儿细菌性肺炎的相关性。方法采用双抗体夹心ELISA法分别测定26例细菌性肺炎患儿（肺炎组）治疗前后和14例无明显气道炎性反应患JL（对照组）BALF中IL-6、IL-8水平，并行中性粒细胞（PMN）和肺泡巨噬细胞（AMs）计数。结果肺炎组治疗前BALF中IL-6和IL-8水平、细胞总数、PMN明显比治疗后及对照组高（P〈0．01）。革兰阴性细菌感染患儿BALF中IL-6、IL-8水平比革兰阳性细菌感染显著升高（P〈0．01）。肺炎组BALF中IL-6水平与AMs呈显著正相关（r=0．7615，P〈0．01），肺炎组BALF中IL-8水平分别与PMN和AMs呈显著正相关（r=0．8956，r=0．6018，P〈0．01）。结论IL-6、IL-8参与小儿细菌性肺炎的炎性反应，BALF中IL-6和IL-8水平动态变化对评价小儿细菌性肺炎的病情有一定的临床意义。     

Cooperation of the inducible nitric oxide synthase and cytochrome P450 1A1 in mediating lung inflammation and mutagenicity induced by diesel exhaust particles

Diesel exhaust particles (DEPs) have been shown to activate oxidant generation by alveolar macrophages (AMs), alter xenobiotic metabolic pathways, and modify the balance of pro-antiinflammatory cytokines. In this study we investigated the role of nitric oxide (NO) in DEP-mediated and DEP organic extract (DEPE) -mediated inflammatory responses and evaluated the interaction of inducible NO synthase (iNOS) and cytochrome P450 1A1 (CYP1A1). Male Sprague-Dawley rats were intratracheally (IT) instilled with saline, DEPs (35 mg/kg), or DEPEs (equivalent to 35 mg DEP/kg), with or without further treatment with an iNOS inhibitor, aminoguanidine (AG; 100 mg/kg), by intraperitoneal injection 30 min before and 3, 6, and 9 hr after IT exposure. At 1 day postexposure, both DEPs and DEPEs induced iNOS expression and NO production by AMs. AG significantly lowered DEP- and DEPE-induced iNOS activity but not the protein level while attenuating DEPE- but not DEP-mediated pulmonary inflammation, airway damage, and oxidant generation by AMs. DEP or DEPE exposure resulted in elevated secretion of both interleukin (IL) -12 and IL-10 by AMs. AG significantly reduced DEP- and DEPE-activated AMs in IL-12 production. In comparison, AG inhibited IL-10 production by DEPE-exposed AMs but markedly increased its production by DEP-exposed AMs, suggesting that NO differentially regulates the pro- and antiinflammatory cytokine balance in the lung. Both DEPs and DEPEs induced CYP1A1 expression. AG strongly inhibited CYP1A1 activity and lung S9 activity-dependent 2-aminoanthracene mutagenicity. These studies show that NO plays a major role in DEPE-induced lung inflammation and CYP-dependent mutagen activation but a lesser role in particulate-induced inflammatory damage.     

Disruption of MicroRNA Expression in Human Airway Cells by Diesel Exhaust Particles Is Linked to Tumorigenesis-Associated Pathways

Background

Particulate matter (PM) is associated with adverse airway health effects; however, the underlying mechanism in disease initiation is still largely unknown. Recently, microRNAs (miRNAs; small noncoding RNAs) have been suggested to be important in maintaining the lung in a disease-free state through regulation of gene expression. Although many studies have shown aberrant miRNA expression patterns in diseased versus healthy tissue, little is known regarding whether environmental agents can induce such changes.

Objectives

We used diesel exhaust particles (DEP), the largest source of emitted airborne PM, to investigate pollutant-induced changes in miRNA expression in airway epithelial cells. We hypothesized that DEP exposure can lead to disruption of normal miRNA expression patterns, representing a plausible novel mechanism through which DEP can mediate disease initiation.

Methods

Human bronchial epithelial cells were grown at air–liquid interface until they reached mucociliary differentiation. After treating the cells with 10 μg/cm² DEP for 24 hr, we analyzed total RNA for miRNA expression using microarray profile analysis and quantitative real-time polymerase chain reaction.

Results

DEP exposure changed the miRNA expression profile in human airway epithelial cells. Specifically, 197 of 313 detectable miRNAs (62.9%) were either up-regulated or down-regulated by 1.5-fold. Molecular network analysis of putative targets of the 12 most altered miRNAs indicated that DEP exposure is associated with inflammatory responses pathways and a strong tumorigenic disease signature.

Conclusions

Alteration of miRNA expression profiles by environmental pollutants such as DEP can modify cellular processes by regulation of gene expression, which may lead to disease pathogenesis.     

白细胞介素-8与子宫内膜异位关系

目的 探讨白细胞介素-8(IL-8)在子宫内膜异位症中(简称内异症)的表达及在发病机制中作用。方法 将内异症患者按修订后的美国生殖协会子宫内膜异位症分期法(R-AFS)进行分期,分别用酶联免疫吸附法(ELISA)检测腹腔液中IL-8表达水平,实时荧光定量PCR检测子宫内膜组织IL-8 mRNA水平,以log互补DNA/log3-磷酸甘油醛脱氢酶(log cDNA/log GAPDH)代表其mRNA水平。结果 内异症患者与对照组腹腔液中IL-8的表达水平分别为(23.37±8.23),(12.32±4.27)pg/ml,差异有统计学意义(P<0.05),内异症增生期和分泌期内膜组织IL-8 mRNA表达水平分别为1.3011±0.2689,1.1239±0.2738,2者差异无统计学意义(P>0.05)。结论 IL-8作为自分泌生长因子和正反馈因子,直接促进内膜基质细胞生长,在内异症的发生和发展中起重要作用。     

Induction of IL-6 and inhibition of IL-8 secretion in the human airway cell line Calu-3 by urban particulate matter collected with a modified method of PM sampling

Exposure to particulate matter (PM) induces inflammatory cytokines. In the present study, we evaluated the secretion of IL-6 and IL-8 by an airway cell line exposed to PM with a mean aerodynamic size equal to or less than 10 or 2.5 μm (PM₁₀ and PM_2.5, respectively) collected in Mexico City, using a modified high-volume sampling method avoiding the use of solvents or introducing membrane components into the samples. PM was collected on cellulose-nitrate (CN) membranes modified for collection on high-volume samplers. Composition of the particles was evaluated by particle-induced X-ray emission (PIXE) and scanning electron microscopy. The particles (10-160 μg/cm²) were tested on Calu-3 cells. Control cultures were exposed to LPS (10 ng/mL to 100 μg/mL) or silica (10-160 μg/cm²). IL-6 and IL-8 secretions were evaluated by ELISA. An average of 10 mg of PM was recovered form each cellulose-nitrate filter. No evidence of contamination from the filter was found. Cells exposed to PM₁₀ presented an increase in the secretion of IL-6 (up to 400%), while IL-8 decreased (from 40% to levels below the detection limit). A similar but weaker effect was observed with PM_2.5. In conclusion, our modified sampling method provides a large amount of urban PM free of membrane contamination. The urban particles induce a decrease in IL-8 secretion that contrasts with the LPS and silica effects. These results suggest that the regulation of IL-8 expression is different for urban particles (complex mixture containing combustion-related particles, soil and biologic components) than for biogenic compounds or pure mineral particles.