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[建立铜绿假单胞菌肺炎中性粒细胞减少大鼠模型及其炎症反应研究]

[Establishing a model of neutropenia rat with Pseudomonas aeruginosa pneumonia and a study on its inflammatory reaction].

作者信息

Li Z, Qu J, He L

机构信息

Department of Pulmonary Medicine, Zhongshan Hospital, Institute of Respiratory Disease, Fudan University, Shanghai 200032, China.

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2001 Nov;24(11):674-8.

Abstract

OBJECTIVE

To establish an animal model and study the inflammatory reaction of P. Aeruginosa pneumonia in neutropenia rats. METHODS Fifty SD rats were randomly divided into two groups: drug-treated group and control CON group. Drug-treated group was given a combination regimen of cyclophosphamide (15 m x kg(-1) d(-1)) and cortisone acetate (100 mg x kg(-1) x d(-1)) for seven days,then both groups were intratracheally challenged with P. Aeruginosa (0.2 ml ATCC 27853 6 x 10(8) CFU/ml). Their blood, bronchial alveolar lavage fluid (BALF) and lung tissue were collected before and 3, 6, 9, 24 h after challenging. Cytological and bacteriological examinations were performed, histopathologic changes of lung tissue were observed. Total proteins (TP) of BALF and the wet/dry ratio (W/D) of lung tissue were determined.

RESULTS

(1) Compared with CON group, rats of drug-treated group showed obviously weight loss and thymus atrophy [(141 +/- 8) g] vs [(201 +/- 14) g], [(0.06 +/- 0.05) g] vs (0.40 +/- 0.10) g, P < 0.001], developed leukocytopenia [(0.9 +/- 0.3) x 10(9)/L] vs [(7.3 +/- 1.9) x 10(9)/L, P < 0.001] and the numbers of alveolar macrophage in BALF decreased significantly; (2) After PA challenging, drug-treated rats showed less activities, worse situations and higher mortality (10.8%) than CON group which recovered quickly and none of them died. PA was identified from samples of BALF and lung tissue, both groups developed inflammatory reaction at 6 - 9 h in high level. Pulmonary pathologic study revealed that polymorphonuclears response of drug-treated group was delayed and less serious than that of CON group [at 9 h, drug-treated group, (102 +/- 13)/HP vs CON group (291 +/- 20)/HP, P < 0.01], however, interstitial edema, capillary congestion and focal hemorrhages were more obvious; (3) After PA challenging especially at 6 - 9 h, W/D ratio and TP concentrations were significantly high in both groups than those of before [drug-treated group, W/D (9.2 +/- 1.3) vs (5.9 +/- 1.4) TP (1.59 +/- 0.83) mg/ml vs (0.19 +/- 0.07) mg/ml; CON group:W/D 7.2 +/- 2.5 vs 4.9 +/- 0.8,TP(0.42 +/- 0.16) mg/ml vs (0.13 +/- 0.04) mg/ml, P < 0.05], however the alterations were much greater in drug-treated group (P < 0.05). Alteration of TP concentration showed some correlation with numbers of polymorphonuclears in lung tissue of CON group (r = 0.926, P < 0.05), but not in drug-treated group (r = 0.58, P = 0.31).

CONCLUSION

It was indicated that there may be other mechanisms than polymorphonuclears infiltration contributing to the more severe lung injury in drug-treated group characterized as neutropenia.

摘要

目的

建立动物模型,研究中性粒细胞减少大鼠铜绿假单胞菌肺炎的炎症反应。方法:将50只SD大鼠随机分为两组:药物治疗组和对照组。药物治疗组给予环磷酰胺(15mg·kg-1·d-1)和醋酸可的松(100mg·kg-1·d-1)联合方案,连续7天,然后两组均经气管内接种铜绿假单胞菌(0.2ml ATCC 27853 6×108CFU/ml)。在接种前及接种后3、6、9、24小时采集它们的血液、支气管肺泡灌洗液(BALF)和肺组织。进行细胞学和细菌学检查,观察肺组织的组织病理学变化。测定BALF中的总蛋白(TP)和肺组织的湿/干比(W/D)。

结果

(1)与对照组相比,药物治疗组大鼠体重明显减轻,胸腺萎缩[(141±8)g对(201±14)g,(0.06±0.05)g对(0.40±0.10)g,P<0.001],出现白细胞减少[(0.9±0.3)×109/L对(7.3±1.9)×109/L,P<0.001],BALF中肺泡巨噬细胞数量显著减少;(2)接种铜绿假单胞菌后,药物治疗组大鼠活动减少,情况较差,死亡率较高(10.8%),而对照组恢复迅速,无一死亡。从BALF和肺组织样本中鉴定出铜绿假单胞菌,两组在6-9小时时均出现高水平的炎症反应。肺病理研究显示,药物治疗组的多形核细胞反应延迟且比对照组轻[9小时时,药物治疗组,(102±13)/HP对对照组(291±20)/HP,P<0.01],然而,间质水肿、毛细血管充血和局灶性出血更明显;(3)接种铜绿假单胞菌后,尤其是在6-9小时时,两组的W/D比和TP浓度均显著高于接种前[药物治疗组,W/D(9.2±1.3)对(5.9±1.4),TP(1.59±0.83)mg/ml对(0.19±0.07)mg/ml;对照组:W/D 7.2±2.5对4.9±0.8,TP(0.42±0.16)mg/ml对(0.13±0.04)mg/ml,P<0.05],然而药物治疗组的变化更大(P<0.05)。TP浓度的变化与对照组肺组织中多形核细胞数量有一定相关性(r=0.926,P<0.05),但在药物治疗组中无相关性(r=0.58,P=0.31)。

结论

表明在以中性粒细胞减少为特征的药物治疗组中,除多形核细胞浸润外,可能还有其他机制导致更严重的肺损伤。

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