IETM对大鼠肝肺综合征形成过程中肺部NOS表达影响的动态研究
发布时间：2019-03-22 14:38
【摘要】： 目的:探讨肠源性内毒素血症对大鼠肝肺综合征(Hepatopulmonary syndrome ,HPS)形成过程中肺部一氧化氮合酶表达的动态变化。 材料与方法:健康雄性Wistar大鼠,体重220-250g,共24只,随机分为4组,正常对照组(control)给予正常饲料和自来水,模型组以掺入胆固醇的玉米面作饲料,胆固醇占饲料重量的0.5%(前2周的每千克饲料中含有200g猪油),以30%酒精作为其唯一饮料。在实验第一天脊背皮下注射CCL4原液,按5ml/kg给予,以后每隔3天皮下注射40% CCL4(用玉米油稀释),按3ml/kg给予,分别于4周末、6周末、8周末处死动物,留取肝组织、肺组织标本、血标本,通过生化、放免、病理、免疫组化、RT-PCR等方法检测相关指标。 结果:1.HPS成过程中所致的肠源性内毒素血症的动态观察结果 在HPS动物模型复制4周起,血浆内毒素水平显著升高,6周达到高峰,8周有所下降,但仍显著高于正常水平。结果提示在HPS形成的过程中伴随存在IETM。 2.HPS形成过程中肺部一氧化氮合酶(nitric oxide synthase,NOS)的动态观察结果 2.1肺组织诱导型一氧化氮合酶(inducible nitric oxide synthase iNOS)和内皮型一氧化氮合酶(endothelial nitric oxide synthase ,eNOS)蛋白表达的动态观察结果,在HPS形成过程中,实验第4周肺部iNOS和eNOS的蛋白表达显著增高,在6周和8周时间点呈明显的持续上升趋势; 2.2肺组织iNOS和eNOS基因表达的动态观察结果,在HPS形成过程中肺部iNOS的基因表达逐渐增加,4周末即有升高,6周末继续升高,到8周末达到高峰;eNOS的基因表达4周起也有增高,到8周达到高峰,两者表达具有相似的规律。 3.TNF-α和ET-1对HPS形成过程中肺部iNOS和eNOS表达影响的结果 TNF-α4周时显著升高(与对照组比较p0.05),6周与4周比有所下降,但仍显著高于正常对照组,8周升高并达到最高峰;ET-1 4周显著下降(p0.05),6周与4周比有所升高,但仍低于对照组,8周继续升高达到最高峰。TNF-α水平与iNOS、eNOS蛋白和基因之间具有显著相关性。 结论:1. HPS形成过程中,肝脏功能改变是肺功能变化的基础; 2. HPS形成过程中伴随发生的IETM,可能是肝功能障碍导致HPS自发形成的关键; 3. HPS形成过程中,IETM可能对肺部eNOS和iNOS的表达发挥重要的调节作用; 4. IETM对肺部eNOS和iNOS在基因和蛋白水平进行的调节,可能与TNF-α和ET-1相关。
[Abstract]:Aim: to investigate the effect of enterogenic endotoxemia on the expression of nitric oxide synthase (NOS) in the lungs during the formation of (Hepatopulmonary syndrome, HPS) in rats with hepatopulmonary syndrome (HPS). Materials and methods: twenty-four healthy male Wistar rats, weighing 220 脳 250 g, were randomly divided into 4 groups. The normal control group (control) was fed with normal feed and tap water, and the model group was fed with corn flour supplemented with cholesterol. Cholesterol accounts for 0.5% of feed weight (200 g lard per kilogram of feed in the first two weeks), with 30% alcohol as its only drink. On the first day of the experiment, CCL4 was injected subcutaneously into the spinal back, given according to 5ml/kg, and then subcutaneously injected with 40% CCL4 (diluted with corn oil) every three days, given according to 3ml/kg. The animals were killed at the end of 4, 6 and 8 weeks, respectively, and the liver tissue was retained. Lung tissue specimens, blood samples, biochemical, radioimmunoassay, pathology, immunohistochemistry, RT-PCR and other methods to detect the related indicators. Results: the results of dynamic observation of enterogenic endotoxemia induced by 1.HPS in HPS animal model increased significantly from 4 weeks after replication, reached a peak at 6 weeks, and decreased at 8 weeks. However, it is still significantly higher than the normal level. The results suggest that there is IETM. in the process of HPS formation. Pulmonary nitric oxide Synthase (nitric oxide synthase, during 2.HPS formation Results 2.The expression of inducible nitric oxide synthase (inducible nitric oxide synthase iNOS) and endothelial nitric oxide synthase (endothelial nitric oxide synthase, eNOS) protein in lung tissue were observed dynamically. In the course of HPS formation, the protein expression of iNOS and eNOS in lung increased significantly at the 4th week, and continued to increase at the 6th and 8th week. 2. The dynamic observation of iNOS and eNOS gene expression in lung tissue showed that the expression of iNOS gene increased gradually during the formation of HPS, increased at the end of 4 weeks, continued to increase at the end of 6 weeks, and reached the peak at the end of 8 weeks. The gene expression of eNOS also increased at 4 weeks and reached its peak at 8 weeks. 3. The effect of TNF-伪 and ET-1 on the expression of iNOS and eNOS in lung during HPS formation. The expression of TNF-伪 increased significantly at 4 weeks (p0.05 as compared with control group), but decreased at 6 weeks and 4 weeks, respectively. However, it was still significantly higher than that of the normal control group, and increased and reached the highest level at 8 weeks. The level of ET-1 decreased significantly at 4 weeks (p0.05), increased at 6 and 4 weeks, but still lower than that of the control group, and reached the highest level at 8 weeks. There was a significant correlation between iNOS,eNOS protein and genes and the level of TNF-伪. Conclusions: 1. During the formation of HPS, the change of liver function is the basis of the change of lung function. The concomitant occurrence of IETM, during the formation of HPS may be the key to spontaneous formation of HPS due to liver dysfunction. During the formation of HPS, IETM may play an important role in regulating the expression of eNOS and iNOS in the lungs. The regulation of IETM on the gene and protein levels of eNOS and iNOS may be related to TNF- 伪 and ET-1.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363
本文编号：2445678
[Abstract]:Aim: to investigate the effect of enterogenic endotoxemia on the expression of nitric oxide synthase (NOS) in the lungs during the formation of (Hepatopulmonary syndrome, HPS) in rats with hepatopulmonary syndrome (HPS). Materials and methods: twenty-four healthy male Wistar rats, weighing 220 脳 250 g, were randomly divided into 4 groups. The normal control group (control) was fed with normal feed and tap water, and the model group was fed with corn flour supplemented with cholesterol. Cholesterol accounts for 0.5% of feed weight (200 g lard per kilogram of feed in the first two weeks), with 30% alcohol as its only drink. On the first day of the experiment, CCL4 was injected subcutaneously into the spinal back, given according to 5ml/kg, and then subcutaneously injected with 40% CCL4 (diluted with corn oil) every three days, given according to 3ml/kg. The animals were killed at the end of 4, 6 and 8 weeks, respectively, and the liver tissue was retained. Lung tissue specimens, blood samples, biochemical, radioimmunoassay, pathology, immunohistochemistry, RT-PCR and other methods to detect the related indicators. Results: the results of dynamic observation of enterogenic endotoxemia induced by 1.HPS in HPS animal model increased significantly from 4 weeks after replication, reached a peak at 6 weeks, and decreased at 8 weeks. However, it is still significantly higher than the normal level. The results suggest that there is IETM. in the process of HPS formation. Pulmonary nitric oxide Synthase (nitric oxide synthase, during 2.HPS formation Results 2.The expression of inducible nitric oxide synthase (inducible nitric oxide synthase iNOS) and endothelial nitric oxide synthase (endothelial nitric oxide synthase, eNOS) protein in lung tissue were observed dynamically. In the course of HPS formation, the protein expression of iNOS and eNOS in lung increased significantly at the 4th week, and continued to increase at the 6th and 8th week. 2. The dynamic observation of iNOS and eNOS gene expression in lung tissue showed that the expression of iNOS gene increased gradually during the formation of HPS, increased at the end of 4 weeks, continued to increase at the end of 6 weeks, and reached the peak at the end of 8 weeks. The gene expression of eNOS also increased at 4 weeks and reached its peak at 8 weeks. 3. The effect of TNF-伪 and ET-1 on the expression of iNOS and eNOS in lung during HPS formation. The expression of TNF-伪 increased significantly at 4 weeks (p0.05 as compared with control group), but decreased at 6 weeks and 4 weeks, respectively. However, it was still significantly higher than that of the normal control group, and increased and reached the highest level at 8 weeks. The level of ET-1 decreased significantly at 4 weeks (p0.05), increased at 6 and 4 weeks, but still lower than that of the control group, and reached the highest level at 8 weeks. There was a significant correlation between iNOS,eNOS protein and genes and the level of TNF-伪. Conclusions: 1. During the formation of HPS, the change of liver function is the basis of the change of lung function. The concomitant occurrence of IETM, during the formation of HPS may be the key to spontaneous formation of HPS due to liver dysfunction. During the formation of HPS, IETM may play an important role in regulating the expression of eNOS and iNOS in the lungs. The regulation of IETM on the gene and protein levels of eNOS and iNOS may be related to TNF- 伪 and ET-1.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R363
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