TNF-α、SP-A在儿童不同程度肺炎中的表达和临床意义

目的：观察不同程度小儿肺炎时,血清致炎性因子肿瘤坏死因子（TNF-α）、肺表面活性蛋白A（SP-A）表达含量的变化及其临床意义。方法将195例肺炎儿童根据病情严重程度将其分为普通肺炎患儿及重症肺炎患儿,另外选择65例非肺炎患儿作为对照组,采用ELISA方法检测不同程度肺炎血清中的TNF-α和 SP-A 水平。结果 TNF-α、SP-A在普通、重症肺炎患儿血清中表达水平显著升高,且重症肺炎组的TNF-α、SP-A表达水平显著高于普通肺炎组。结论在小儿重症肺炎患儿发病过程中,血清中肿瘤坏死因子TNF-α和肺表面活性蛋白SP-A的表达水平明显增多,可以反映肺损伤程度,可作为临床检测指标。     

SP-A在急性肺损伤各组织含量变化及相关性研究

目的：通过检测急性肺损伤大鼠肺及大肠等组织SP-A在肺与大肠等组织含量变化并分析其相关性,从而为中医“肺与大肠相表里”理论提供实验依据。方法：本实验采用脂多糖气道内滴入制备急性肺损伤动物模型,应用ELISA方法分别检测急性肺损伤及其药物干预后SP-A在肺及大肠等组织的含量,并进行相关性分析。结果：随着急性肺损伤加重,SP-A在肺、大肠、肺泡灌洗液及血清中含量逐渐降低,药物干预后能逆转这种趋势,呈一致性变化。各组织间SP-A含量的相关性比较,治疗前肺与大肠相关系数r=0.521,治疗后肺与大肠相关系数r=0.615。结论：SP-A在肺和大肠组织的含量变化相关性最密切,从而为中医“肺与大肠相表里”理论提供实验依据。     

Amniotic fluid concentration of surfactant proteins in intra-amniotic infection

Objective. Pulmonary surfactant is a complex molecule of lipids and proteins synthesized and secreted by type II alveolar cells into the alveolar epithelial lining. Both lipid and protein components are essential for lung function in postnatal life. Infection is a well-established cause of preterm delivery, and several inflammatory cytokines play a role in the mechanisms of preterm parturition. An increased concentration of inflammatory cytokines in amniotic fluid or fetal plasma has been linked to the onset of preterm parturition and fetal/neonatal injury, including cerebral palsy and chronic lung disease. Experimental evidence indicates that inflammatory mediators also regulate surfactant protein synthesis, and histologic chorioamnionitis is associated with a decreased incidence of hyaline membrane disease in neonates. This study was conducted to determine if amniotic fluid concentrations of surfactant protein (SP)-A, SP-B, and SP-D change in patients with and without intra-amniotic infection (IAI).

Materials and methods. A case–control study was conducted to determine amniotic fluid concentrations of SP-A, SP-B, SP-D, and total protein in patients who had an amniocentesis performed between 18 and 34 weeks of gestation for the detection of IAI in patients with spontaneous preterm labor with intact membranes (n = 42) and cervical insufficiency prior to the application of cerclage (n = 6). Amniotic fluid samples were selected from a bank of biological specimens and included patients with (n = 16) and without (n = 32) IAI matched for gestational age at amniocentesis. Intra-amniotic infection was defined as a positive amniotic fluid culture for microorganisms. Each group was further subdivided according to a history of corticosteroid administration within 7 days prior to amniocentesis into the following subgroups: (1) patients without IAI who had received antenatal corticosteroids (n = 21), (2) patients with IAI who had received antenatal corticosteroids (n = 9), (3) patients without IAI who had not received antenatal corticosteroids (n = 11), and (4) patients with IAI who had not received antenatal corticosteroids (n = 7). Amniotic fluid was obtained by transabdominal amniocentesis. SP-A, SP-B, and SP-D concentrations in amniotic fluid were determined by enzyme-linked immunosorbent assay (ELISA). Non-parametric statistics were used for analysis.

Results. Women with IAI had a higher median amniotic fluid concentration of SP-B and of SP-B/total protein, but not other SPs, than those without IAI (both p = 0.03). Among patients who had received antenatal corticosteroids, the median amniotic fluid concentration of SP-B and of SP-B/total protein was significantly higher in patients with IAI than in those without IAI (SP-B, IAI: median 148 ng/mL, range 37.3–809 ng/mL vs. without IAI: median 7.2 ng/mL, range 0–1035 ng/mL; p = 0.005 and SP-B/total protein, IAI: median 14.1 ng/mg, range 4.3–237.5 ng/mg vs. without IAI: median 1.45 ng/mg, range 0–79.5 ng/mg; p = 0.003). Among women who had not received antenatal corticosteroids, the median amniotic fluid concentrations of SP-B and of SP-B/total protein were not significantly different between patients with and without IAI (SP-B, IAI: median 4 ng/mL, range 0–31.4 ng/mL vs. without IAI: median 3.4 ng/mL, range 0–37 ng/mL; p = 0.8 and SP-B/total protein, IAI: median 0.55 ng/mg, range 0–6.96 ng/mg vs. without IAI: median 0.59 ng/mg, range 0–3.28 ng/mg; p = 0.9). The median amniotic fluid concentrations of SP-A, SP-A/total protein, SP-D, and SP-D/total protein were not significantly different between patients with and without IAI whether they received antenatal corticosteroids or not (all p > 0.05).

Conclusions. IAI was associated with an increased amniotic fluid concentration of SP-B in patients who received antenatal corticosteroids within 7 days prior to amniocentesis.     

Surfactant phosphatidylcholine metabolism in neonates with meconium aspiration syndrome

OBJECTIVE: Because meconium directly inhibits surfactant function, we sought to determine the effect of meconium on endogenous surfactant synthesis and clearance. STUDY DESIGN: We studied surfactant phosphatidylcholine kinetics with the use of stable isotopes in 11 newborn infants with meconium aspiration syndrome (MAS) who required extracorporeal membrane oxygenation (ECMO). For comparison we studied 6 neonates with persistent pulmonary hypertension (PPHN) on ECMO and 10 term neonates ventilated for non-pulmonary indications and not on ECMO. All patients received a 24-hour [U- 13C]glucose infusion as precursor for the palmitic acid in surfactant phosphatidylcholine. RESULTS: In the meconium group, the maximal 13C-incorporation in phosphatidylcholine (PC) was half of that in controls (0.09 +/- 0.01 vs 0.18 +/- 0.03 atom percent excess [APE], P = .027). There was a trend toward lower surfactant synthesis in the MAS group (3.3 +/- 0.7%/day) and PPHN group (2.6 +/- 0.3%/day) compared with controls 8.0 +/- 2.4%/day, P = .058). Significantly lower PC concentrations in tracheal aspirates were found in the MAS group (4.4 +/- 2.6 mg/mL) and PPHN group (3.6 +/- 2.0 mg/mL) compared with controls (12.8 +/- 2.6 mg/mL, P = .01). Endogenously synthesized surfactant had a similar half-life in all groups, ranging from 63 to 98 hours. CONCLUSION: We conclude that surfactant synthesis is disturbed and that surfactant PC concentrations are low in infants with MAS on ECMO.     

羊膜腔内注射肺表面活性物质对宫内感染胎兔肺超微结构及表面活性蛋白A表达的影响

目的 观察羊膜腔内注射肺表面活性物质(PS)对胎膜早破宫内感染之胎兔肺组织超微结构及表面活性蛋白A(surfactant protein A,SP-A)表达的影响.方法 建立胎膜早破宫内感染动物模型,在孕24,26 d给子宫内注射大肠杆菌,分为细菌组和羊膜腔内注射PS组,干预后19.5 h利用电镜观察肺超微结构变化.并分别用免疫组化法及Westernblot(WB)法检测胎肺组织内SP-A表达,同时设立正常对照组.结果 正常胎兔24、26 d处于胎肺发育囊泡期,肺上皮细胞结构完整,可见典型Ⅰ型、Ⅱ型肺泡上皮细胞;细菌组和PS组胎兔肺泡内、支气管管腔内、胞质内均可见到杆菌菌体、浸润的炎症细胞;与对照组相比,细菌组Ⅱ型肺泡上皮细胞数量减少,板层体呈空泡化改变,有不同程度的凋亡;PS组Ⅱ型肺泡上皮细胞数量、板层小体数量较细菌组和对照组明显增多,且呈不同形式出现,细胞发生凋亡少.细菌组SP-A免疫组化及蛋白表达均降低,与对照组相比差异具有显著性(P<0.05);PS组SP-A表达升高,与对照组差异无显著性(P>0.05).结论 宫内感染后肺泡超微结构发生改变,SP-A表达下降;而在感染条件下羊膜腔内注射PS可改善肺泡超微结构的变化,并上调SP-A表达,对肺成熟有明显促进作用.     

结核性和肺腺癌性胸腔积液中肺表面活性物质相关蛋白A、D的检测分析

目的探讨胸水中肺表面活性物质相关蛋白－A、D联合对原发性肺腺癌性与结核性胸腔积液的鉴别诊断价值。方法收集36例原发性肺腺癌性和28例结核性胸腔积液患者的胸水,采用 western blotting检测胸水中肺表面活性物质相关蛋白-A、D。结果原发性肺腺癌和结核性胸膜炎患者胸水中SP A光密度值分别为265.1±141.16和180.0±64.1;原发性肺腺癌和结核性胸膜炎患者胸水中SP-D光密度值分别为299.9±140.6和206.8±86.8。36例原发性肺腺癌患者中有23例胸水中SP-A＞250,有16例SP-D＞250,而SP-A＞250和（或）SP-D＞250有29例。28例结核性胸膜炎患者胸水中SP-A、SP-D均未超过250。结论肺表面活性物质相关蛋白A对结核性胸膜炎与原发性肺腺癌所致胸腔积液鉴别诊断有临床价值,联合肺表面活性物质相关蛋白D对两者的鉴别诊断更有意义。     

平喘固金颗粒对COPD肺脾肾三虚模型大鼠的实验研究

目的 探讨平喘固金颗粒对慢性阻塞性肺疾病(COPD)肺脾肾三虚证模型大鼠肺功能、肺表面活性物质SP-A、IL-8、肺泡灌洗液Ⅰ、Ⅲ型胶原、右心室肥厚指标及肺组织病理变化的影响.方法 60只SD健康大鼠随机分为空白对照组,模型组,金水宝胶囊组,平喘固金颗粒大、中、小剂量组(10.0、5.0、2.5g/kg),用气管内滴入脂多糖及烟熏,并配合番泻叶泻下法、背部注射醋酸氢化可的松,建立肺脾肾三虚证COPD大鼠模型,测定各组大鼠肺功能、肺表面活性物质SP-A、IL-8、肺泡灌洗液Ⅰ、Ⅲ型胶原、右心室肥厚指标,HE染色观察大鼠肺组织病理变化.结果 平喘固金颗粒大剂量组FVC、FEV0.3、FEV0.3/FVC、PEF与模型组比较显著增加(P<0.05);平喘固金颗粒大剂量组显著降低IL-8和Ⅲ型胶原含量,升高SP-A含量,对Ⅰ型胶原无明显影响(P<0.05);平喘固金颗粒大剂量组能显著降低大鼠右心室肥厚指标(P<0.05);模型组大鼠支气管肺组织HE染色结果符合COPD病理改变,与模型组比较,平喘固金颗粒中剂量组炎症吸收良好.结论 平喘固金颗粒可明显改善肺脾肾三虚型COPD大鼠肺功能;显著降低IL-8和Ⅲ型胶原含量;升高SP-A含量;显著降低大鼠右心肥厚指标;减轻肺组织病理学改变.     

A case of interstitial pneumonia caused by bucillamine: a study using serological markers

Abstract

The patient was a 61-year-old man diagnosed with rheumatoid arthritis (RA) in 2001. He initially received treatment at a nearby clinic, but his condition could not be satisfactorily controlled. He subsequently consulted our hospital during the same year. Although his symptoms improved in response to treatment at our hospital, slight fever, cough, and then high fever and dyspnea subsequently developed. A diagnosis of interstitial pneumonia was made on the basis of findings of diagnostic imaging. The time course of changes in serological markers, including surfactant protein A (SP-A), surfactant protein D (SP-D), and KL-6, as well as markers of inflammatory reaction and lactate dehydrogenase was examined to determine the clinical significance of serological markers in the management of interstitial pneumonia.     

人参皂甙对内毒素所致大鼠急性肺损伤的影响

目的:探讨人参皂甙对内毒素所致大鼠急性肺损伤的保护作用及作用机制。方法:将成年大鼠30只按雌雄对半分为3组(每组10只):正常对照组(A组)、内毒素组(B组)、人参皂甙组(C组)。股静脉注射LPS(5mg/kg)复制大鼠ALI模型,比较各组大鼠的一般状况,观测各组大鼠肺脏的大体形态,测定动脉血氧分压(PaO2),氧合指数(PaO2/FiO2),肺湿重/干重(W/D),做病理切片观察肺组织学改变及免疫组织化学法检测肺组织中SP-A蛋白水平的表达情况。结果:与A组比较,B组的PaO2,PaO2/FiO2显著降低(P<0.01),W/D显著增高(P<0.01);C组的PaO2,PaO2/FiO2相对降低(P<0.05),W/D相对增高(P<0.05),B组和C组间PaO2,PaO2/FiO2和W/D存在显著差异(P<0.01)。免疫组化半定量结果显示,A组大鼠肺组织中SP-A蛋白呈强阳性表达。B组与A组相比,明显减弱(P<0.01)。C组与B组相比,B组大鼠肺组织中SP-A蛋白明显增强,呈阳性(P<0.01)。结论:人参皂甙可能通过上调SP-A蛋白的表达保护急性肺损伤。     

Role of surfactant protein-A (SP-A) in lung injury in response to acute ozone exposure of SP-A deficient mice

Millions are exposed to ozone levels above recommended limits, impairing lung function, causing epithelial damage and inflammation, and predisposing some individuals to pneumonia, asthma, and other lung conditions. Surfactant protein-A (SP-A) plays a role in host defense, the regulation of inflammation, and repair of tissue damage. We tested the hypothesis that the lungs of SP-A(-/-) (KO) mice are more susceptible to ozone-induced damage. We compared the effects of ozone on KO and wild type (WT) mice on the C57BL/6 genetic background by exposing them to 2 parts/million of ozone for 3 or 6 h and sacrificing them 0, 4, and 24 h later. Lungs were subject to bronchoalveolar lavage (BAL) or used to measure endpoints of oxidative stress and inflammation. Despite more total protein in BAL of KO mice after a 3 h ozone exposure, WT mice had increased oxidation of protein and had oxidized SP-A dimers. In KO mice there was epithelial damage as assessed by increased LDH activity and there was increased phospholipid content. In WT mice there were more BAL PMNs and elevated macrophage inflammatory protein (MIP)-2 and monocyte chemoattractant protein (MCP)-1. Changes in MIP-2 and MCP-1 were observed in both KO and WT, however mRNA levels differed. In KO mice MIP-2 mRNA levels changed little with ozone, but in WT levels they were significantly increased. In summary, several aspects of the inflammatory response differ between WT and KO mice. These in vivo findings appear to implicate SP-A in regulating inflammation and limiting epithelial damage in response to ozone exposure.