目的 动态观察产前炎症暴露生后大鼠肺结缔组织生长因子(CTGF)表达和肺泡结构变化,探讨CTGF对肺发育的影响机制。方法 Wistar孕大鼠随机分为实验组和对照组,分别于孕19 d和20 d腹腔注射脂多糖(LPS2.5 mg/kg)和等量生理盐水,足月自然分娩后,于日龄1、3、7、14 d两组分别取8只仔鼠,麻醉处死后取肺组织,左肺行HE染色,观察肺组织结构,测量肺泡数量、肺泡间隔厚度、肺泡占组织面积比例,右肺采用Real time PCR和Western blot方法测定肺组织中CTGF mRNA及蛋白表达水平。结果 1)随日龄增加,两组大鼠肺泡数量、肺泡占组织面积比逐渐增加,肺泡间隔变薄。生后1 d、3 d、7 d及14 d,实验组肺泡数量明显少于对照组(P<0.05或<0.01);生后1 d、3 d和7 d,实验组肺泡占组织面积比明显大于对照组(P<0.01);生后1 d及3 d,实验组肺泡间隔厚度明显小于对照组(P=0.000)。2)生后1 d、3 d和7 d实验组CTGFmRNA表达明显高于对照组(P<0.05或<0.01)。3)生后1、3、7、 14 d实验组CTGF蛋白表达均明显强于对照组(P均<0.05)。结论 产前感染导致生后大鼠肺CTGF表达增加与肺结构异常有关。
Abstract
Objective To explore the role for connective tissue growth factor(CTGF) in the lung development by dynamic observation of morphology and the expression of CTGF in lungs of rat pups exposed to antenatal inflammation. Methods Pregnant Wistar rats were randomly divided into experimental group and control group.Pregnant rats of both groups were intraperitoneal injected with either lipopolysaccharide ( LPS)2.5 mg/kg or the same volume of normal saline on embryonic day 19 and 20,respectively,and were allowed to term deliver.On postnatal days 1,3,7 and 14 (D1,D3,D7 and D14),eight pups of each group were killed by 10% chloral hydrate (1 ml/kg) and lungs were collected.Left lungs of each group were used to assess lung histological changes after hematoxylin and eosin(HE) staining by measuring the mean numbers of alveolar,the mean ratio of alveolar surface area to per tissue and the mean thickness of alveolar septum.Right lungs of each group were used to measure the expression of CTGF mRNA and protein by Real-time polymerase chain reaction (Real-time PCR) and Western blot. Results 1)With the increasing of postnatal days,the mean numbers of alveolar and the mean ratio of alveolar surface area to per tissue in both groups increased,the mean thickness of alveolar septum got thinner.On D1,D3,D7 and D14,the mean alveolar numbers of the experimental group were significantly less than those of the control group (P<0.05 or <0.01).On D1,D3 and D7,the mean ratios of alveolar surface area to per tissue were significantly larger than those of control group(P<0.01).On D1 and D3,the alveolar septum thickness was significantly thinner than that in control group(P=0.000).2)On D1,D3,andD7,the expression of CTGF mRNA of the experimental group were significantly higher than those of the control group(P<0.05 or <0.01).3)On D1,D3,D7 and D14,the expression of CTGF protein of the experimental group were significantly higher than those of the control group(P all<0.05). Conclusion Antenatal infection can up-regulate expression of CTGF,which may assiociate with disorders of lung development.
关键词
产前感染 /
肺发育 /
结缔组织生长因子 /
大鼠 /
支气管肺发育不良
Key words
antenatal infection /
lung development /
connective tissue growth factor /
rat /
bronchopulmonary dysplasia
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基金
辽宁省自然基金资助课题(201202273)
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