唾液乳杆菌对哮喘Balb/c小鼠气道高反应性及T-bet/GATA-3表达的影响

目的探讨唾液乳杆菌对哮喘Balb/c小鼠气道高反应性及T-bet/GATA-3表达的影响。方法选择30只4周、体质量16~18 g、SPF级Balb/c雌性小鼠,随机分成3组:正常对照组(N)、哮喘组(A)、哮喘加唾液乳杆菌组(AH)。应用卵蛋白建立急性哮喘模型,无创肺功能仪测定小鼠气道反应性、病理HE染色观察肺部炎症改变、Real-time PCR法检测肺组织中GATA-3 m RNA及T-bet m RNA表达。结果哮喘组小鼠较正常对照组小鼠气道反应性增高,AH组较哮喘组小鼠气道反应性降低(P0.05);病理HE染色见哮喘组支气管管壁增厚、管腔狭窄、气管及血管周围可见大量以嗜酸性粒细胞为主的炎症细胞浸润,管腔中较多炎性分泌物,AH组小鼠肺组织病理改变较哮喘组明显减轻;哮喘组小鼠肺组织中GATA-3m RNA表达水平较N组明显升高,T-betm RNA表达水平较N组明显降低(P0.05);而AH组小鼠肺组织中GATA-3m RNA表达水平较哮喘组明显降低,T-betm RNA表达水平较哮喘组明显升高(P0.05)。结论致敏前唾液乳杆菌灌胃一定程度上减轻了哮喘小鼠的气道高反应性及气道炎症,通过在转录水平增加T-betm RNA表达同时抑制GATA-3m RNA表达,进而改善哮喘小鼠Th1/Th2失衡,为支气管哮喘的免疫防治提供新的途径。     

神经激肽1受体拮抗剂WIN62577对大鼠气道平滑肌细胞增殖和迁移的影响

目的 探讨神经激肤1受体拮抗剂 WIN62577对大鼠气道平滑肌细胞(ASMC)增殖和迁移的影响.方法 原代培养大鼠ASMC,将传代纯化的第4代细胞随机分为空白对照组、白细胞介素13(IL-13)干预组和WIN62577+IL-13干预组.MTT分析各组细胞的生长曲线;流式细胞仪分析各组G1、S和G2期细胞比例变化;Transwell小室检测WIN62577对ASMC迁移的影响.结果 IL-13干预组与空白对照组、WIN62577+IL-13干预组比较,48和72 h ASMC增殖的差异均有统计学意义(P<0.05).WIN62577+IL-13干预组ASMC大多停留于G1期,与空白对照组比较同期细胞比例差异均有统计学意义(P<0.05).经WIN63577干预后迁移的ASMC数量显著减少(P<0.05).结论 WIN62577有抑制ASMC增殖和迁移的作用.     

Change of alveolar epithelial type Ⅱ cells and pulmonary surfactant protein A in young rats with acute lung injury

Objective To study the temporal changes of alveolar epithelial type Ⅱ cells and surfactant pro-tein A in young rats with acute lung injury induced by lipopolysaecharide. Method Totally 110 SD young rats (male:53, female : 57) were randomly divided into ALI and normal control groups (six subgroups in each group).LPS(4 mg/kg) was given intraperitoneally in ALI group. The same amount of normal saline was given in the con-trol groups. Eight rats in each subgroup were sacrificed at 6, 12, 24, 36, 48 and 72 hours after the injection.Lung samples were taken for transmission electron microscope examination. RT-PCR was epmloyed for the mea-surement of SP-A mRNA. Western blot was used for the detection of SP-A in the lung tissue. ANOVA and homo-geneity of variance test were performed by SPSS 12.0. Results The microvilli disappeared at 24 hours after the injection of LPS. The number of lamellar body (LBs) was provisionality increased at 24 hours and 48 hours. The ring-like an'angement of LBs around nucleus and the giant LB with vacuole-like deformity were found as the main characteristics of AEC- Ⅱ in ALI at 48 hours. The number of LBs reduced and broken and residual LB remained at 72 hours. SP-A elevated greatly from 24 to 48 hours (P < 0.01), reached peak at 36 hours (6.94 ± 0.80, P <0.01),reached the lowest level(3.87 ±0.50, P <0.01)at 72 hours. Conclusions The pathological changes of AEC-Ⅱ and SP-A in lung tissue wiht ALI are time-dependent. The typical alterations of AEC- Ⅱ occurs at 48 hours accompanied by the compensatory increase of SP-A. AEC- Ⅱ is seriously injuried with the typical changes of LBs and the diminishing of SP-A in lung tissue.     

急性肺损伤幼鼠肺泡Ⅱ型上皮细胞和肺表面活性蛋白A的变化

目的 探索急性肺损伤(ALI)时肺泡Ⅱ型上皮细胞(AEC-Ⅱ)和肺表面活性蛋白A(SP-A)的变化规律.方法 110只SD幼鼠(雄性53只,雌性57只)随机分为对照组、ALI组(每组分6个亚组).ALI组腹腔注射脂多糖(LP3,4 mg/ks),对照组注射生理盐水.于注射后6,12,24,36,48,72 h每组处死8只幼鼠并取材.透射电镜下观察各组AEC-Ⅱ超微结构的变化.采用半定量RT-PCR法测量各组肺组织SP-A mRNA的表达,用Western blot法测量肺组织SP-A含量.采用SPSS 12.0统计软件包进行方差分析和方差齐性检验.结果 注射LPS 24 h后,AEC-Ⅱ表面微绒毛消失.24 h和48 h时AEC-Ⅱ中板层小体(LB)出现暂时性数馈增加.48 h时LB呈特征性"指环状"绕核排列和巨大的空泡样LB.72 h时LB数目显著减少,可见破碎和残余的LB.SP-A从24 h至48 h表达显著增强(P<0.01),于36 h达最高值(6.94±0.80,P<0.01),72 h 下降至最低点(3.87 ±0.50,P<0.01).结论 ALI时AEC-Ⅱ形态变化与肺组织SP-A含量变化是时间依赖性的.AEC-Ⅱ损伤早期伴有肺组织SP-A的代偿性增加.AEC-Ⅱ损伤加重时LB出现特征性变化,可能与SP-A的代偿有关.ALI晚期,AEC-Ⅱ的严重损伤与SP-A的下降密切相关.     

γ-维生素E对哮喘小鼠血清及肺泡灌洗液嗜酸性粒细胞趋化因子的影响

目的 探讨γ-维生素E(γ-T)对哮喘的治疗作用及机制。方法 将40只雄性Balb/c小鼠(4周,18~20 g)随机分为正常组、哮喘组、地塞米松组及γ-T治疗组。给予小鼠腹腔注射及雾化吸入卵白蛋白建立哮喘气道炎症反应模型,分别给予地塞米松及γ-T,正常组仅给予生理盐水。28 d后处死小鼠,留取血清及肺泡灌洗液。ELISA法测定各组血清及肺泡灌洗液中嗜酸性粒细胞趋化因子(eotaxin)的含量。用one-way ANOVA检验组间差异,P＜0.05具有统计学意义。结果 哮喘组小鼠血清及肺泡灌洗液中eotaxin浓度均较正常组升高,地塞米松组及γ-T组均较哮喘组下降。在血清中地塞米松组下降最明显,γ-T组次之,γ-T组与哮喘组,γ-T组与地塞米松组相比差异无统计学意义(P>0.05);在BALF中,γ-T组的eotaxin下降最明显,与哮喘组相比,二者间差异明显(P<0.05);γ-T组与地塞米松组相比,二者差异无统计学意义(P>0.05)。 结论 γ-T有利于减少哮喘小鼠肺组织eotaxin,γ-T可能是有用的抗哮喘药物。     

瘦素及其受体在哮喘BALB/c小鼠肺组织中的表达及布地奈德的干预作用

目的 探讨不同严重程度哮喘小鼠及布地奈德干预后肺组织中瘦素及其受体表达的变化规律。方法 将40 只Balb/c 小鼠随机分成对照组、诱喘3 d 组、诱喘7 d 组和布地奈德干预组, 每组10 只。采用卵清蛋白(OVA)致敏后分别激发3 d 和7 d 建立小鼠哮喘模型;布地奈德干预组于激发前1 h 雾化吸入布地奈德混悬液干预;对照组不予任何处理。苏木精- 伊红染色观察各组小鼠气道炎症情况;免疫组化、Western blot 和Real-time PCR 方法检测肺组织中的瘦素及其受体蛋白和mRNA 的表达。结果 哮喘组气道病理学改变较对照组、布地奈德干预组明显, 而哮喘组中, 诱喘7 d 组改变较诱喘3 d 组明显。肺组织中瘦素蛋白及mRNA 表达在诱喘3 d 组较对照组显著增高(P<0.01), 诱喘7 d 组较3 d 组显著增高(P<0.01);瘦素受体蛋白表达在诱喘3 d组较对照组显著降低(P<0.01), 布地奈德干预组较诱喘7 d 组明显增高(P<0.01);瘦素受体mRNA 表达在4组间比较差异无统计学意义(P>0.05)。结论 哮喘肺组织中瘦素呈高表达, 而瘦素受体呈低表达;布地奈德可以上调瘦素受体的表达, 而对瘦素表达无明显影响。     

Change of alveolar epithelial type Ⅱ cells and pulmonary surfactant protein A in young rats with acute lung injury

Objective To study the temporal changes of alveolar epithelial type Ⅱ cells and surfactant pro-tein A in young rats with acute lung injury induced by lipopolysaecharide. Method Totally 110 SD young rats (male:53, female : 57) were randomly divided into ALI and normal control groups (six subgroups in each group).LPS(4 mg/kg) was given intraperitoneally in ALI group. The same amount of normal saline was given in the con-trol groups. Eight rats in each subgroup were sacrificed at 6, 12, 24, 36, 48 and 72 hours after the injection.Lung samples were taken for transmission electron microscope examination. RT-PCR was epmloyed for the mea-surement of SP-A mRNA. Western blot was used for the detection of SP-A in the lung tissue. ANOVA and homo-geneity of variance test were performed by SPSS 12.0. Results The microvilli disappeared at 24 hours after the injection of LPS. The number of lamellar body (LBs) was provisionality increased at 24 hours and 48 hours. The ring-like an'angement of LBs around nucleus and the giant LB with vacuole-like deformity were found as the main characteristics of AEC- Ⅱ in ALI at 48 hours. The number of LBs reduced and broken and residual LB remained at 72 hours. SP-A elevated greatly from 24 to 48 hours (P < 0.01), reached peak at 36 hours (6.94 ± 0.80, P <0.01),reached the lowest level(3.87 ±0.50, P <0.01)at 72 hours. Conclusions The pathological changes of AEC-Ⅱ and SP-A in lung tissue wiht ALI are time-dependent. The typical alterations of AEC- Ⅱ occurs at 48 hours accompanied by the compensatory increase of SP-A. AEC- Ⅱ is seriously injuried with the typical changes of LBs and the diminishing of SP-A in lung tissue.     

Change of alveolar epithelial type Ⅱ cells and pulmonary surfactant protein A in young rats with acute lung injury

Objective To study the temporal changes of alveolar epithelial type Ⅱ cells and surfactant pro-tein A in young rats with acute lung injury induced by lipopolysaecharide. Method Totally 110 SD young rats (male:53, female : 57) were randomly divided into ALI and normal control groups (six subgroups in each group).LPS(4 mg/kg) was given intraperitoneally in ALI group. The same amount of normal saline was given in the con-trol groups. Eight rats in each subgroup were sacrificed at 6, 12, 24, 36, 48 and 72 hours after the injection.Lung samples were taken for transmission electron microscope examination. RT-PCR was epmloyed for the mea-surement of SP-A mRNA. Western blot was used for the detection of SP-A in the lung tissue. ANOVA and homo-geneity of variance test were performed by SPSS 12.0. Results The microvilli disappeared at 24 hours after the injection of LPS. The number of lamellar body (LBs) was provisionality increased at 24 hours and 48 hours. The ring-like an'angement of LBs around nucleus and the giant LB with vacuole-like deformity were found as the main characteristics of AEC- Ⅱ in ALI at 48 hours. The number of LBs reduced and broken and residual LB remained at 72 hours. SP-A elevated greatly from 24 to 48 hours (P < 0.01), reached peak at 36 hours (6.94 ± 0.80, P <0.01),reached the lowest level(3.87 ±0.50, P <0.01)at 72 hours. Conclusions The pathological changes of AEC-Ⅱ and SP-A in lung tissue wiht ALI are time-dependent. The typical alterations of AEC- Ⅱ occurs at 48 hours accompanied by the compensatory increase of SP-A. AEC- Ⅱ is seriously injuried with the typical changes of LBs and the diminishing of SP-A in lung tissue.     

2009年流行性甲型H1N1流感重症肺炎患儿血清肺表面活性蛋白A、B、C、D的变化

目的探讨流行性甲型H1N1流感（简称甲流）重症肺炎患儿肺表面活性蛋白A、B、C、D在血清含量变化的特点,寻找肺组织损伤的生化指标。方法采用ELISA方法检测8例甲流重症肺炎患儿（甲流组）和20例健康儿童（对照组）血清肺表面活性蛋白A、B、C、D含量,同时化验前白蛋白和白蛋白等。结果甲流组血清肺表面活性蛋白A、B、C、D含量高于对照组（P均〈0．05）;外周血白细胞、血浆前白蛋白、白蛋白等低于对照组（P均〈0．05）;二聚体高于对照组（P〈0．05）。结论甲流重症肺炎患儿血清肺表面活性蛋白A、B、C、D含量可作为肺组织损伤严重程度的检验指标。     

Change of alveolar epithelial type Ⅱ cells and pulmonary surfactant protein A in young rats with acute lung injury

Objective To study the temporal changes of alveolar epithelial type Ⅱ cells and surfactant pro-tein A in young rats with acute lung injury induced by lipopolysaecharide. Method Totally 110 SD young rats (male:53, female : 57) were randomly divided into ALI and normal control groups (six subgroups in each group).LPS(4 mg/kg) was given intraperitoneally in ALI group. The same amount of normal saline was given in the con-trol groups. Eight rats in each subgroup were sacrificed at 6, 12, 24, 36, 48 and 72 hours after the injection.Lung samples were taken for transmission electron microscope examination. RT-PCR was epmloyed for the mea-surement of SP-A mRNA. Western blot was used for the detection of SP-A in the lung tissue. ANOVA and homo-geneity of variance test were performed by SPSS 12.0. Results The microvilli disappeared at 24 hours after the injection of LPS. The number of lamellar body (LBs) was provisionality increased at 24 hours and 48 hours. The ring-like an'angement of LBs around nucleus and the giant LB with vacuole-like deformity were found as the main characteristics of AEC- Ⅱ in ALI at 48 hours. The number of LBs reduced and broken and residual LB remained at 72 hours. SP-A elevated greatly from 24 to 48 hours (P < 0.01), reached peak at 36 hours (6.94 ± 0.80, P <0.01),reached the lowest level(3.87 ±0.50, P <0.01)at 72 hours. Conclusions The pathological changes of AEC-Ⅱ and SP-A in lung tissue wiht ALI are time-dependent. The typical alterations of AEC- Ⅱ occurs at 48 hours accompanied by the compensatory increase of SP-A. AEC- Ⅱ is seriously injuried with the typical changes of LBs and the diminishing of SP-A in lung tissue.