肝肺综合征大鼠模型肺血管内巨噬细胞iNOS和HO-1的表达

首都医科大学学报 ›› 2009, Vol. 30 ›› Issue (2): 222-226.

肝肺综合征大鼠模型肺血管内巨噬细胞iNOS和HO-1的表达

高君¹, 王宇², 张忠涛², 李建设², 马雪梅², 赵丽珍³

1. 首都医科大学附属北京朝阳医院京西院区肝胆胰脾外科;2. 首都医科大学附属北京友谊医院普外科;3. 包头医学院第一附属医院消化内科

收稿日期:2008-02-18 修回日期:1900-01-01 出版日期:2009-04-21 发布日期:2009-04-21
通讯作者: 王宇

Expression of Inducible Nitric Oxide Synthase and Heme Oxygenase-1 in Pulmonary Intravascular Macrophages of Rats with Hepatopulmonary Syndrome

GAO Jun¹, WANG Yu², ZHANG Zhong-tao², LI Jian-she², MA Xue-mei², ZHAO Li-zhen³

1. Department of Hepatobilary Surgery, Jingxi Area of Beijing Chaoyang Hospital, Capital Medical University;2. Department of General Surgery, Beijing Friendship Hospital, Capital Medical University;3. Department of Gastroenterology, First Hospital Affiliated to Baotou Medical University

Received:2008-02-18 Revised:1900-01-01 Online:2009-04-21 Published:2009-04-21

摘要/Abstract

摘要： 目的探讨肺血管内巨噬细胞(pulmonary intravascular macrophages,PIM)在肝肺综合征(hepatopulmonary syndrome,HPS)发病机制中的作用。方法用随机数字表法将Sprague-Dawley(SD)大鼠随机分为对照组和CCl₄组。模型制备后取动脉血作血气分析,即静脉血测内毒素和肝功能。行肝、肺病理检查,取肠系膜淋巴结作细菌培养。大鼠巨噬细胞单抗免疫组化染色观察PIM的黏附、聚集情况。免疫组化染色检测诱导型一氧化氮合酶(iNOS)和血红素氧合酶-1(HO-1)的蛋白表达。SYBR Green Ⅰ实时定量PCR方法检测肺组织中iNOS和HO-1的mRNA表达。结果对照组肺泡细胞无变性坏死,肺泡形态正常,无炎性细胞浸润、间质水肿、纤维增生和透明膜形成。CCl₄组肺泡毛细血管增生、扩张,肺泡间隔增宽、容量减小。CCl₄组PaO₂(81.39±2.36)mmHg和PaCO₂(30.55±2.87)mmHg较对照组PaO₂(98.99±3.41)mmHg和PaCO₂(36.26±2.88)mmHg,差异有统计学意义(P<0.05)。CCl₄组肺泡-动脉血氧分压(A-aDO₂) (30.79±5.73)mmol/L较对照组A-aDO₂(3.79±0.86)mmol/L,差异有统计学意义(P<0.05)。CCl₄组内毒素(0.30±0.18)ZU/L较对照组内毒素(0.05±0.02)ZU/L,差异有统计学意(P<0.05)。对照组的肠系膜淋巴结培养未见细菌生长,CCl₄组肠系膜淋巴结培养阳性率为62.5%。对照组肺血管内无巨噬细胞聚集,CCl₄组肺血管内大量巨噬细胞聚集,并且iNOS和HO-1蛋白表达均定位于PIM内。CCl₄组iNOS(0.03±0.01)和HO-1(0.16±0.04)的mRNA表达明显高于对照组iNOS(0.01±0.01)和HO-1(0.07±0.05)(P<0.05)。结论 PIM黏附、聚集,分泌iNOS和HO-1可能是NO和CO合成增多,是引起HPS的重要因素。

关键词: 肝肺综合征, 肺血管内巨噬细胞, 诱导型一氧化氮合酶, 血红素氧合酶-1

Abstract: Objective To explore the role of pulmonary intravascular macrophages(PIM ) in the pathogenesis of hepatopulmonary syndrome(HPS). Methods Sprague-Dawley(SD) rats were divided into two test groups, control group and CCl₄ group. Arterial blood was collected for measurement of blood gases, venous blood for hepatic function and endotoxin levels. Pathology of liver and lung was done. The mesenteric lymph nodes were dissected for bacteriology studies. Immunolocalization of macrophages was performed using monoclonal macrophage antibody ED1 on paraffin sections. Proteins of iNOS and HO-1 of lung tissue were examined by immunohistochemistry. By real-time polymerase chain reaction(PCR) using SYBR Green Ⅰ, the expression levels of iNOS and HO-1 mRNA in lung tissues were measured. Results There was no inflammation, edema, fibrosis, alveolar collapse and hyaline membrane formation in the lung control group. All the lungs from CCl₄ group showed widened alveolar wall architectures, widespread dilatation of alveolar capillaries and decreased alveolar volume. PaO₂(81.39±2.36)mmHg and PaCO₂(30.55±2.87)mmHg decreased significantly in CCl₄ group compared with(98.99±3.41)mmHg and(36.26±2.88)mmHg in the control group(P<0.05). Alveolar-arterial oxygen difference(A-aDO₂) increased significantly in CCl₄ group(30.79±5.73) as compared with the control(3.79±0.86) (P<0.05). Endotoxin level increased significantly in CCl₄ group(0.30±0.13)ZU/L compared with control(0.05±0.02)ZU/L(P<0.05). Culture-positive mesenteric lymph nodes were found in 62.5% of CCl₄ group. There were not Culture-positive mesenteric lymph nodes in the control. All lungs from CCl₄ group showed accumulation of large mononuclear macrophage-like cells within the lumen of numerous small muscular and nonmuscular pulmonary vessels. Using immunohistochemical analysis, iNOS and HO-1 expressions were localized to PIM in CCl₄ group. The mRNA expression of iNOS(0.03±0.01) and HO-1 (0.16±0.04) increased significantly in CCl₄ group than(0.01±0.01) and(0.07±0.02) in the control(P<0.05). Conclusion Cirrhosis results in bacterial translocation from the gastrointestinal tract and endotoxemia. High concentrations of endotoxin in the pulmonary bloodstream activate PIM. PIM aggregates and expresses iNOS and HO-1. Thus the increasing amount of NO and CO released in the lungs results in HPS.

Key words: hepatopulmonary syndrome, pulmonary intravascular macrophages, inducible nitric oxide synthase, heme oxygenase-1

中图分类号:

R 575.3

高君;王宇;张忠涛;李建设;马雪梅;赵丽珍. 肝肺综合征大鼠模型肺血管内巨噬细胞iNOS和HO-1的表达[J]. 首都医科大学学报, 2009, 30(2): 222-226.

GAO Jun;WANG Yu;ZHANG Zhong-tao;LI Jian-she;MA Xue-mei;ZHAO Li-zhen. Expression of Inducible Nitric Oxide Synthase and Heme Oxygenase-1 in Pulmonary Intravascular Macrophages of Rats with Hepatopulmonary Syndrome[J]. Journal of Capital Medical University, 2009, 30(2): 222-226.

[1]	闫峰, 尹洁, 罗玉敏, 李森, 王玉兰, 赵咏梅. 远隔缺血预适应对大鼠局灶性脑缺血后诱导型一氧化氮合酶的影响[J]. 首都医科大学学报, 2014, 35(3): 315-319.
[2]	王会玲;周晓春;马春虎;王冰;王晓民. 雷公藤内酯醇对Aβ_1-42激活的小胶质细胞一氧化氮释放和诱导型一氧化氮合酶表达的影响[J]. 首都医科大学学报, 2012, 33(5): 643-646.
[3]	江瑛;闫丽;王红霞;芦玲巧;曾翔俊;朱盈芬;张立克. 11,12-环二十碳三烯酸对心脏缺血/再灌注损伤大鼠血红素氧合酶-1的影响[J]. 首都医科大学学报, 2007, 28(1): 64-67.