Inner Shell of the Chestnut (Castanea crenatta) Suppresses Inflammatory Responses in Ovalbumin-Induced Allergic Asthma Mouse Model

The inner shell of the chestnut (Castanea crenata) contains various polyphenols, which exert beneficial biological effects. Hence, we assessed the anti-inflammatory efficacy of a chestnut inner shell extract (CIE) in ovalbumin (OVA)-induced allergic asthma. We intraperitoneally injected 20 μg of OVA with 2 mg of aluminum hydroxide on days 0 and 14. On test days 21, 22, and 23, the mice were treated with aerosolized 1% (w/v) OVA in saline. CIE was administered orally at 100 and 300 mg/kg on days 18–23. CIE significantly reduced inflammatory cytokines and cells and immunoglobulin-E increased by OVA. Anti-inflammatory efficacy was revealed by reduction of inflammatory cell migration and mucus secretion in lung tissue. Further, CIE suppressed the OVA-induced nuclear factor kappa B (NF-κB) phosphorylation. Accordingly, the expression of cyclooxygenase (COX-2), inducible nitric oxide synthase (iNOS), and matrix metalloproteinase-9 (MMP-9) were decreased sequentially in lung tissues. CIE alleviated OVA-induced airway inflammation by restraining phosphorylation of NF-κB and the sequentially reduced expression of iNOS, COX-2, leading to reduced MMP-9 expression. These results indicate that CIE has potential as a candidate for alleviating asthma.     

大鼠哮喘模型肺组织血红素氧合酶-1蛋白及基因表达

目的 探讨大鼠哮喘模型肺组织血红素氧合酶-1(HO-1)蛋白及基因表达特点以及其与HO-1的活性之间的相关性.方法 清洁级雄性SD大鼠20只,随机分为对照组和哮喘组,每组10只.测定全血碳氧血红蛋白(COHb)的百分比含量;计算肺泡灌洗液(BALF)沉渣中细胞总数和分类;计数气道壁中浸润的炎性细胞数;用免疫组织化学染色法观察HO-1在哮喘大鼠肺组织的表达;用RT-PCR方法 检测哮喘大鼠肺组织HO-1基因表达.结果 HO-1阳性细胞主要定位于支气管上皮细胞及黏膜下巨噬细胞,气道平滑肌细胞偶有表达.2组表达HO-1的阳性细胞百分比分别为(5.03±1.22)%和(27.14±4.68)%;2组HO-1基因表达光密度比值分别为(0.32±0.05),(0.68±0.02);2组全血COHb的百分比含量分别为(0.45±0.35),(3.89±1.15);哮喘组HO-1蛋白、基因的表达水平以及全血COHb的百分比含量均显著高于正常组(P＜0.01).HO-1蛋白表达水平、基因表达水平与全血COHb的百分比含量均呈正相关(r=0.897,0.971,P＜0.01).结论 哮喘大鼠的肺组织HO-1蛋白基因表达水平显著增加,且反应HO-1活性的全血COHb的百分比含量亦显著增加,提示HO-1可能参与了哮喘的发病过程.     

The Effects of Microbial Materials Adhered to Asian Sand Dust on Allergic Lung Inflammation

Asian sand dust (ASD) containing microbiological materials, sulfate (SO₄²), and nitrate (NO₃⁻) derived from air pollutants in East China, reportedly cause adverse respiratory health effects. ASD aggravates ovalbumin (OVA)-associated experimental lung eosinophilia. In this study, the toxic materials adsorbed onto ASD were excluded by heat treatment at 360°C for 30 min. The effects of nonheated ASD or heated ASD (H-ASD) toward the allergic lung inflammation were compared in murine lungs. ICR mice were administered intratracheally with normal saline (control), H-ASD, ASD, OVA, OVA + H-ASD, and OVA + ASD, four times at 2-week intervals. ASD only increased neutrophils in bronchoalveolar lavage fluids (BALFs) along with pro-inflammatory mediators, such as keratinocyte chemoattractant (KC). H-ASD and ASD enhanced eosinophil recruitment induced by OVA in the alveoli and in the submucosa of the airway, which has a goblet cell proliferation in the bronchial epithelium. The two ASDs synergistically increased interleukin-5 (IL-5), monocyte chemotactic protein-3 (MCP-3), and eotaxin, which were associated with OVA, in BALF. The enhancing effects were much greater in ASD than in H-ASD. The two ASDs induced the adjuvant effects to specific IgE and IgG1 production by OVA. In the in vitro study using RAW264.7 cells, ASD increased the expression of Toll-like receptor 2 (TLR 2) mRNA but not TLR4 mRNA. H-ASD caused no expression of either TLR mRNA. These results suggest that the aggravated lung eosinophilia by ASD may be due to activation of Th2-associated immune response via the activation of TLR2 by microbial components adhered to ASD.     

平喘汤对哮喘大鼠气道转化生长因子-β1 mRNA和Ⅲ型胶原表达的影响

目的:探讨平喘汤对哮喘大鼠模型气道壁厚度及转化生长因子-β1 mRNA(TGF-β1 mRNA)和Ⅲ型胶原表达的影响及可能的作用机制。方法:将40只雄性Wistar大鼠随机分为正常对照组、哮喘组、地塞米松组、平喘汤组,每组10只。用卵白蛋白(OVA)致敏吸入激发制备哮喘模型。在哮喘激发4周后测量肺组织的TGF-β1 mRNA和气道壁Ⅲ型胶原的表达。结果:模型组TGF-β1mRNA、Ⅲ型胶原表达明显高于正常对照组(P<0.05);平喘汤组和地塞米松组TGF-β1 mRNA、Ⅲ型胶原表达显著低于模型组(P<0.05);气道壁厚度模型组明显高于正常对照组(P<0.05);平喘汤组和地塞米松组显著低于模型组(P<0.05)。结论:平喘汤通过降低TGF-β1 mRNA的过度表达,抑制Ⅲ型胶原的沉积,减轻气道重塑的发生。     

胸腺和活化调节趋化因子在哮喘大鼠中的作用及布地奈德干预的影响

目的 探讨胸腺和活化调节趋化因子(thymus and activation regulated chemokine, TARC)在哮喘大鼠中的作用及吸入布地奈德干预对TARC的影响。方法 30只SD大鼠采用随机区组设计分为正常对照组、哮喘组及布地奈德干预组。以卵清白蛋白(ovalbumin, OVA)致敏激发法制备大鼠哮喘急性模型, 干预组于14 d雾化吸入布地奈德进行干预。于最后1次雾化后24 h, 尾静脉取血, 支气管肺泡灌洗留取灌洗液(bronchoalveolar lavage fluid, BALF), 留取右肺组织标本。行血清和BALF中细胞计数, 应用酶联免疫吸附(ELISA)法测定血清和BALF中上清液中嗜酸粒细胞(Eos)、IL-4及TARC的水平。采用免疫组化法测定肺组织中TARC及IL-4蛋白的表达。结果 1) 哮喘组大鼠血清及BALF中的Eos、IL-4和TARC水平均高于对照组(P<0.05);肺组织中IL-4及TARC的蛋白表达也高于对照组(P<0.05);布地奈德干预后上述各项指标值均低于哮喘组(P<0.05), 布地奈德干预组的各项指标与对照组相比差异无统计学意义(P>0.05)。2)哮喘组大鼠肺组织中TARC的蛋白表达与IL-4呈正相关(r=0.50, P<0.05)。结论 哮喘大鼠中TARC含量明显增高, 吸入布地奈德可降低TARC的水平。提示TARC在哮喘的发病中起重要的作用, 下调TARC可能是激素治疗哮喘的一个重要机制。     

高迁移率族蛋白B1激活JAK2/STAT3并促进支气管上皮细胞分泌哮喘相关炎症因子

目的 探讨高迁移率族蛋白B1(HMGB1)参与调节支气管上皮细胞分泌哮喘相关炎症因子的机制。方法 20只雌性BALB/c小鼠随机分对照组和哮喘组,采用卵清蛋白腹腔注射致敏联合雾化吸入激发建立哮喘小鼠模型。支气管上皮细胞16-HBE分组处理:空白对照组,HMGB1组(浓度分别为100、200、500 ng/mL),500 ng/mL HMGB1+AG490组,500 ng/mL HMGB1+雷帕霉素组。HE染色观察小鼠气道结构;qRT-PCR和免疫组化法检测两组小鼠肺部组织HMGB1的表达;ELISA法测定两组小鼠支气管肺泡灌洗液(BALF)上清中HMGB1的含量和各组细胞上清液中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)和白细胞介素-1α(IL-1α)水平。Western blot检测各组细胞中p-JAK2和p-STAT3的蛋白水平。结果 哮喘组小鼠HMBG1的表达比正常小鼠明显升高。HMGB1升高细胞中TNF-α、IL-6和IL-1α水平,上调p-JAK2和p-STAT3水平,并且呈现浓度依赖性关系。AG490和雷帕霉素处理抑制细胞中p-JAK2和p-STAT3表达,同时降低TNF-α、IL-6和IL-1α水平。结论 HMGB1参与调控哮喘慢性炎症反应,可能与活化JAK2/STAT3信号通路促进支气管上皮细胞分泌炎症因子有关。     

布地奈德对哮喘大鼠肺组织病理改变及TRAF2表达的影响

【目的】 观察哮喘大鼠肺组织病理改变和TRAF2表达的水平,探讨布地奈德治疗哮喘的可能作用机制。 【方法】 采用大鼠哮喘模型,随机分成哮喘组、对照组和布地奈德组,HE染色观察肺组织炎性改变,免疫组织化学法检测肺组织TRAF2的表达。 【结果】 哮喘组大鼠的毛发、体重、活动度等一般情况改变和肺组织炎性改变较对照组有显著变化,布地奈德能明显减轻上述改变。哮喘组(0.317±0.041 OD值)支气管壁TRAF2的光密度值显著高于对照组(0.220±0.057 OD值)(P＜0.01);布地奈德组支气管壁(0.236±0.033 OD值)TRAF2的光密度值显著低于哮喘组(P＜0.01),与对照组相比差异无统计学意义(P>0.05),但肺组织中其表达水平却显著高于哮喘组。 【结论】 哮喘大鼠TRAF2的表达水平增强,它可能参与了哮喘的气道炎症过程;布地奈德能减轻气道炎症,其机制可能是部分通过TRAF2途经。     

罗格列酮对非酒精性脂肪性肝炎大鼠的保护作用

目的 探讨罗格列酮对高脂饮食性非酒精性脂肪性肝炎(NASH)大鼠的保护作用及其部分机制.方法 30只雄性SD大鼠以掷硬币法均分为正常组、模型组和罗格列酮预防组,模型组和罗格列酮预防组高脂喂养12周制备NASH模型,罗格列酮预防组同时每天予4 mg/kg罗格列酮灌胃.观察肝大体、肝指数和病理学变化;ELISA方法检测血清肿瘤坏死因子-α、前列腺素E2;免疫组织化学方法观察肝过氧化物酶体增殖物激活受体γ (PPARγ)、核因子(NF) -κB和环氧合酶(COX)-2的表达,荧光定量PCR和Western blot检测肝COX-2基因和蛋白表达变化.结果 和正常组比较,模型组肝指数升高(3.92±0.72比5.71±1.05,P=0.004),肝脂肪变、炎症和纤维化改变明显,血清肿瘤坏死因子-α(11.72±2.47比29.39±5.32,P=0.002)、前列腺素E2(236.60±24.90比288.24±17.17,P=0.004)水平升高,罗格列酮预防组肝指数、肝脂肪变、炎症和纤维化以及血清肿瘤坏死因子-α、前列腺素E2水平明显好转.免疫组织化学结果显示模型组肝PPARγ表达下降、NF-κB和COX-2表达升高,定量PCR和Western blot结果显示模型组肝COX-2表达较正常组升高(0.57±0.08比2.83±0.24,P=0.0007; 0.38±0.03比1.00±0.03,P=0.004).和模型组比较,罗格列酮预防组PPARγ表达升高、NF-κB和COX-2(基因:2.83±0.24比0.46±0.11,P=0.002;蛋白:1.00±0.03比0.62±0.02,P=0.006)表达下降.结论 罗格列酮可激活PPARγ后抑制NF-κB和COX-2表达,从而减轻氧化应激和胰岛素抵抗,预防NASH发生发展.     

Risk of Asthmatic Episodes in Children Exposed to Sulfur Dioxide Stack Emissions from a Refinery Point Source in Montreal,Canada

Background

Little is known about the respiratory effects of short-term exposures to petroleum refinery emissions in young children. This study is an extension of an ecologic study that found an increased rate of hospitalizations for respiratory conditions among children living near petroleum refineries in Montreal (Canada).

Methods

We used a time-stratified case–crossover design to assess the risk of asthma episodes in relation to short-term variations in sulfur dioxide levels among children 2–4 years of age living within 0.5–7.5 km of the refinery stacks. Health data used to measure asthma episodes included emergency department (ED) visits and hospital admissions from 1996 to 2004. We estimated daily levels of SO₂ at the residence of children using a) two fixed-site SO₂ monitors located near the refineries and b) the AERMOD (American Meteorological Society/Environmental Protection Agency Regulatory Model) atmospheric dispersion model. We used conditional logistic regression to estimate odds ratios associated with an increase in the interquartile range of daily SO₂ mean and peak exposures (31.2 ppb for AERMOD peaks). We adjusted for temperature, relative humidity, and regional/urban background air pollutant levels.

Results

The risks of asthma ED visits and hospitalizations were more pronounced for same-day (lag 0) SO₂ peak levels than for mean levels on the same day, or for other lags: the adjusted odds ratios estimated for same-day SO₂ peak levels from AERMOD were 1.10 [95% confidence interval (CI), 1.00–1.22] and 1.42 (95% CI, 1.10–1.82), over the interquartile range, for ED visits and hospital admissions, respectively.

Conclusions

Short-term episodes of increased SO₂ exposures from refinery stack emissions were associated with a higher number of asthma episodes in nearby children.     

Effect of sulfur dioxide on mucociliary activity and ciliary beat frequency in guinea pig trachea

Summary The effects of 30 min exposure to sulfur dioxide on mucociliary activity (MCA) and ciliary beat frequency (CBF) were studied in 31 guinea pig tracheas. MCA was measured by recording the light reflected from ciliated mucous membranes using an infrared bar code reader. CBF of single ciliated cells obtained by brushing was measured with phase-contrast microscopy. Each tracheal sample was exposed to SO₂ at concentrations ranging from 2.5 to 12.5 ppm, or to air for control purposes. MCA and CBF were measured before and immediately after gas exposure. A reduction in mean MCA of 63% (P = 0.0007) and statistically insignificant changes in CBF (P > 0.05) were recorded at concentrations of 2.5 PPM SO₂. Higher SO₂ concentrations caused a further impairment of MCA as well as a dose-dependent decrease in CBF (P = 0.002). A concentration of 12.5 PPM SO₂ induced a decrease from baseline values of approximately 80% in mean MCA and of roughly 70% in mean CBE This study demonstrates a dose-dependent SO₂-induced decrease in MCA of guinea pig tracheas. The decrease in MCA was associated with an impairment of CBF only at SO₂ concentrations higher than 5.0 ppm.     

Cytotoxic effects of sulfuric acid mist, carbon particulates, and their mixtures on hamster tracheal epithelium.

Hamster tracheal tissue was used to study the effects of H₂SO₄ mist, carbon particles, and mixtures of the two on cilia beating frequency and morphological alterations of respiratory epithelium. Hamsters were exposed for 3 hr to 1.1 mg/m³ H₂SO₄ (mean size, 0.12 μm) and 1.5 mg/m³ carbon (mean size, 0.3 μm) particle aerosols alone or in combination. Trachea of animals exposed in vivo to the mixture and held in vivo showed cytotoxic effects in the epithelium that were greater than those produced by either the acid mist or carbon alone. In tracheas of hamsters exposed in vivo and maintained in vitro the damage produced by acid mist—carbon mixture did not differ significantly from that produced by acid mist per se but was greater than that observed after exposure to carbon. Organ cultures of tracheal rings exposed for 3 hr to a 1:10⁶ dilution of concentrated H₂SO₄ and 100 μg/ml carbon produced epithelial damage in vitro similar to that seen in in vivo exposures. The extent of recovery over a period of 72 hr was also studied following different combinations of in vivo and in vitro exposure and/or maintenance.     

Murine Lung Responses to Ambient Particulate Matter: Genomic Analysis and Influence on Airway Hyperresponsiveness

Background

Asthma is a complex disease characterized by airway hyperresponsiveness (AHR) and chronic airway inflammation. Epidemiologic studies have demonstrated that exposures to environmental factors such as ambient particulate matter (PM), a major air pollutant, contribute to increased asthma prevalence and exacerbations.

Objective

We investigated pathophysiologic responses to Baltimore, Maryland, ambient PM (median diameter, 1.78 μm) in a murine model of asthma and attempted to identify PM-specific genomic/molecular signatures.

Methods

We exposed ovalbumin (OVA)-sensitized A/J mice intratracheally to PM (20 mg/kg), and assayed both AHR and bronchoalveolar lavage (BAL) on days 1, 4, and 7 after PM exposure. Lung gene expression profiling was analyzed in OVA- and PM-challenged mice.

Results

Consistent with this murine model of asthma, we observed significant increases in airway responsiveness in OVA-treated mice, with PM exposure inducing significant changes in AHR in both naive mice and OVA-induced asthmatic mice. PM evoked eosinophil and neutrophil infiltration into airways, elevated BAL protein content, and stimulated secretion of type 1 T helper (T_H1) cytokines [interferon-γ, interleukin-6 (IL-6), tumor necrosis factor-α] and T_H2 cytokines (IL-4, IL-5, eotaxin) into murine airways. Furthermore, PM consistently induced expression of genes involved in innate immune responses, chemotaxis, and complement system pathways.

Conclusion

This study is consistent with emerging epidemiologic evidence and indicates that PM exposure evokes proinflammatory and allergic molecular signatures that may directly contribute to the asthma susceptibility in naive subjects and increased severity in affected asthmatics.     

Lactobacillus fermentum Suo Attenuates HCl/Ethanol Induced Gastric Injury in Mice through Its Antioxidant Effects

The purpose of the study was to determine the inhibitory effects of Lactobacillus fermentum Suo (LF-Suo) on HCl/ethanol induced gastric injury in ICR (Institute for Cancer Research) mice and explain the mechanism of these effects through the molecular biology activities of LF-Suo. The studied mice were divided into four groups: healthy, injured, LF-Suo-L and LF-Suo-H group. After the LF-Suo intragastric administration, the gastric injury area was reduced compared to the injured group. The serum MOT (motilin), SP (substance P), ET (endothelin) levels of LF-Suo treated mice were lower, and SS (somatostatin), VIP (vasoactive intestinal peptide) levels were higher than the injured group mice. The cytokine IL-6 (interleukin 6), IL-12 (interleukin 12), TNF-α (tumor necrosis factor-α) and IFN-γ (interferon-γ) serum levels were decreased after the LF-Suo treatment. The gastric tissues SOD (superoxide dismutase), GSH-Px (glutathione peroxidase), NO (nitric oxide) and activities of LF-Suo treated mice were increased and MDA (malondialdehyde) activity was decreased compared to the injured group mice. By the RT-PCR assay, LF-Suo raised the occludin, EGF (epidermal growth factor), EGFR (epidermal growth factor receptor), VEGF (vascular endothelial growth factor), Fit-1 (fms-like tyrosine kinase-1), IκB-α (inhibitor kappaB-α), nNOS (neuronal nitric oxide synthase), eNOS (endothelial nitric oxide synthase), Mn-SOD, Cu/Zn-SOD, CAT (catalase) mRNA or protein expressions and reduced the COX-2, NF-κB (nuclear factor kappaB), and iNOS (inducible nitric oxide synthase) expressions in gastric tissues compared to the gastric injured group mice. A high concentration (1.0 × 10⁹ CFU/kg b.w.) of LF-Suo treatment showed stronger anti-gastric injury effects compared to a low concentration of (0.5 × 10⁹ CFU/kg b.w.) of LF-Suo treatment. LF-Suo also showed strong survival in pH 3.0 man-made gastric juice and hydrophobic properties. These results indicate that LF-Suo has potential use as probiotics for its gastric injury treatment effects.     

Adjuvant effect of Panax notoginseng saponins on the immune responses to ovalbumin in mice

In this study, the haemolytic activities of Panax notoginseng saponins (PNS) and its adjuvant potentials on the cellular and humoral immune responses of ICR mice against ovalbumin (OVA) were evaluated. We determined the haemolytic activity of PNS using 0.5% rabbit red blood cell. PNS showed a slight haemolytic effect, with its haemolytic percents being 11.59 and 3.60% at the concentrations of 500 and 250 microg/ml, respectively. Furthermore, the adjuvant potential of PNS at three dose levels on the cellular and humoral immune responses of ICR mice against ovalbumin were investigated. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing aluminum hydroxide gel (Alum) (200 microg), Quil A (10 and 50 microg) or PNS (50, 100 or 200 microg) on days 1 and 15. Two weeks later (day 28), concanavalin A (Con A)-, pokeweed (PWM)- and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. PNS significantly enhanced the Con A-, PWM-, and OVA-induced splenocyte proliferation in the OVA-immunized mice at a dose of 100 microg (P < 0.05 or P < 0.025). OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were significantly enhanced by PNS compared with OVA control group (P < 0.025). Moreover, enhancing effect of PNS on the OVA-specific IgG2b antibody responses to OVA in mice were more significant than that of Quil A (P < 0.025). In conclusion, the results suggest that PNS could be safely used as adjuvant with low or non-haemolytic effect.     

Acute effects of inhaled SO2 plus NaCl droplet aerosol on pulmonary function in asthmatic adolescents

Nine adolescent subjects with a diagnosis of extrinsic asthma were exposed at rest for 60 min to the following experimental modes: (a) filtered air; (b) 1 ppm of sulfur dioxide (SO₂) and 1 mg/m³ of sodium chloride (NaCl) droplet aerosol; c 1 mg/m³ of NaCl droplet aerosol alone. All exposures were at 75% relative humidity and 22°C temperature. Functional measurements of total respiratory resistance (R_T), maximal flow at 50 and 75% of expired vital capacity ($\text{V}\text{\_.}{}_{\mathrm{<mtext>max50</mtext>}}$ and $\text{V}\text{\_.}{}_{\mathrm{<mtext>max75</mtext>}}$), forced expiratory volume in 1 sec (FEV_1.0) and functional residual capacity (FRC) were recorded before, during, and after exposures. Significant decreases in $\text{V}\text{\_.}{}_{\mathrm{<mtext>max50</mtext>}}$ and $\text{V}\text{\_.}{}_{\mathrm{<mtext>max75</mtext>}}$ were seen when the group was exposed to the SO₂ plus NaCl droplet aerosol. No significant changes were seen during exposure to filtered air or NaCl droplet aerosol alone. The results suggest that the site of effect of the SO₂NaCl droplet aerosol was in small airways. The asthmatic adolescents appeared to be more sensitive to the SO₂ droplet aerosol than were healthy nonsmoking adults previously studied.     

白三烯受体基因mRNA表达水平与儿童哮喘的相关性

目的:检测了白三烯受体基因mRNA在儿童外周血白细胞中的表达;研究轻、中、重度哮喘患者白三烯受体基因mRNA表达水平是否存在不同;探讨性别和年龄因素对哮喘组白三烯受体基因mRNA表达水平产生的影响。方法:应用逆转录聚合酶链式反应(RT-PCR)技术,对比并定量分析CysLT1-Receptor和CysLT2-Receptor基因mRNA在哮喘组和对照组之间的表达差异。结果:哮喘组及对照组中均有CysLT1-Receptor和CysLT2-Receptor基因mRNA的表达;哮喘组白三烯受体基因mRNA的表达显著增高,(P均<0·05);轻、中、重度哮喘患儿CysLT1-Receptor和CysLT2-Receptor基本mRNA表达水平无显著性差异;性别、年龄因素对哮喘组白三烯受体基因mRNA表达无明显影响P>0·05。结论:CysLT1-Receptor和CysLT2-Receptor基因在儿童外周血白细胞中存在表达,其mRNA表达水平在哮喘组显著增高。性别和年龄因素对哮喘组白三烯受体基因mRNA的表达无明显影响。     

COX-2和VEGF在子宫腺肌病组织中的表达及其意义

目的:观测子宫腺肌病异位子宫内膜组织COX-2和VEGF表达及其临床意义。方法:34例经病理证实为子宫腺肌病标本为研究组,30例非子宫腺肌病子宫标本为对照组。采用免疫组化法检测研究组原位内膜和腺肌病病灶、对照组内膜和子宫平滑肌层中COX-2和VEGF的表达。采用RT-PCR检测COX-2mRNA和VEGFmRNA的表达。结果:COX-2和VEGF在研究组原位内膜和腺肌病病灶中的表达率均高于对照组内膜,研究组腺肌病病灶表达率均高于原位内膜(P<0.01)。增殖期和分泌期二者在腺肌病病灶表达率均无统计学差异(P>0.05)。分泌期二者在研究组和对照组中内膜的表达均高于增殖期(P<0.05)。COX-2和VEGF蛋白表达存在线性相关(r=0.787,0.562,P<0.01)。RT-PCR检测表明,COX-2mRNA在研究组腺肌病病灶存在高表达,对照组肌层和内膜不存在COX-2mRNA表达。比较对照组正常肌层,腺肌病病灶中VEGF165和VEGF121呈现高表达(P<0.01)。结论:COX-2和VEGF的高表达可能与子宫腺肌病的发生、发展有关。     

Preparation and evaluation of antigen/N-trimethylaminoethylmethacrylate chitosan conjugates for nasal immunization

The frequent outbreak of respiratory infectious diseases such as influenza and pulmonary tuberculosis calls for new immunization strategies with high effectiveness. Nasal immunization is one of the most potential methods to prevent the diseases infected through the respiratory tract. In this study, we designed a water-soluble system based on antigen/N-trimethylaminoethylmethacrylate chitosan conjugates for nasal immunization. N-trimethylaminoethylmethacrylate chitosan (TMC) was synthesized by free radical polymerization of chitosan and N-trimethylaminoethylmethacrylate chloride and identified by ¹H NMR and FT-IR. Thiolated ovalbumin (OVA) was covalently conjugated to maleimide modified TMC with high conjugation efficiency. OVA conjugated TMC (OVA–TMC) significantly increased uptake of OVA by Raw 264.7 cells, which was 2.38 times higher than that of OVA/TMC physical mixture (OVA + TMC) at 4 h. After nasal administration, OVA–TMC showed higher transport efficiency to superficial and deep cervical lymph nodes than OVA + TMC or OVA alone. Balb/C mice were intranasally given with OVA–TMC three times at 2-week internals to evaluate the immunological effect. The serum IgG, IgG1 and IgG2a levels of the OVA–TMC group were 17.9–87.9 times higher than that of the OVA + TMC group and comparable to that of the intramuscular group. The secretory IgA levels in nasal wash and saliva of the OVA–TMC group were 5.2–7.1 times higher than that of the OVA + TMC group while the secretory IgA levels of the intramuscular alum-precipitated OVA group were not increased. After immunofluorescence staining of nasal cavity, IgA antibody secreting cells were mainly observed in the lamina propria regions and glands of nasal mucosa. OVA–TMC showed little toxicity to the nasal epithelia or cilia of rats after nasal administration for three consecutive days. These results demonstrated that antigen conjugated TMC can induce both systemic and mucosal immune responses after nasal administration and may serve as a convenient, safe and effective vaccine for preventing respiratory infectious diseases.     

Asian sand dust enhances ovalbumin-induced eosinophil recruitment in the alveoli and airway of mice

Asian sand dust (ASD) containing sulfate (SO4(2-)) reportedly causes adverse respiratory health effects but there is no experimental study showing the effect of ASD toward allergic respiratory diseases. The effects of ASD and ASD plus SO4(2-) toward allergic lung inflammation induced by ovalbumin (OVA) were investigated in this study. ICR mice were administered intratracheally with saline; ASD alone (sample from Shapotou desert); and ASD plus SO4(2-) (ASD-SO4); OVA+ASD; OVA+ASD-SO4. ASD or ASD-SO4 alone caused mild nutrophilic inflammation in the bronchi and alveoli. ASD and ASD-SO4 increased pro-inflammatory mediators, such as Keratinocyte chemoattractant (KC) and macrophage inflammatory protein (MIP)-1 alpha, in bronchoalveolar lavage fluids (BALF). ASD and ASD-SO4 enhanced eosinophil recruitment induced by OVA in the alveoli and in the submucosa of the airway, which has a goblet cell proliferation in the bronchial epithelium. However, a further increase of eosinophils by addition of SO4(2-) was not observed. The two sand dusts synergistically increased interleukin-5 (IL-5) and monocyte chemotactic protein-1 (MCP-1), which were associated with OVA, in BALF. However, the increased levels of IL-5 were lower in the OVA+ASD-SO4 group than in the OVA+ASD group. ASD caused the adjuvant effects to specific-IgG1 production by OVA, but not to specific-IgE. These results suggest that the enhancement of eosinophil recruitment in the lung is mediated by synergistically increased IL-5 and MCP-1. IgG1 antibodies may play an important role in the enhancement of allergic reaction caused by OVA and sand dust. However, extra sulfate may not contribute to an increase of eosinophils.