IETM對大鼠肝肺綜合征形成過程中肺部NOS表達影響的動態(tài)研究
發(fā)布時間：2019-03-22 14:38
【摘要】： 目的:探討腸源性內(nèi)毒素血癥對大鼠肝肺綜合征(Hepatopulmonary syndrome ,HPS)形成過程中肺部一氧化氮合酶表達的動態(tài)變化。 材料與方法:健康雄性Wistar大鼠,體重220-250g,共24只,隨機分為4組,正常對照組(control)給予正常飼料和自來水,模型組以摻入膽固醇的玉米面作飼料,膽固醇占飼料重量的0.5%(前2周的每千克飼料中含有200g豬油),以30%酒精作為其唯一飲料。在實驗第一天脊背皮下注射CCL4原液,按5ml/kg給予,以后每隔3天皮下注射40% CCL4(用玉米油稀釋),按3ml/kg給予,分別于4周末、6周末、8周末處死動物,留取肝組織、肺組織標本、血標本,通過生化、放免、病理、免疫組化、RT-PCR等方法檢測相關(guān)指標。 結(jié)果:1.HPS成過程中所致的腸源性內(nèi)毒素血癥的動態(tài)觀察結(jié)果 在HPS動物模型復制4周起,血漿內(nèi)毒素水平顯著升高,6周達到高峰,8周有所下降,但仍顯著高于正常水平。結(jié)果提示在HPS形成的過程中伴隨存在IETM。 2.HPS形成過程中肺部一氧化氮合酶(nitric oxide synthase,NOS)的動態(tài)觀察結(jié)果 2.1肺組織誘導型一氧化氮合酶(inducible nitric oxide synthase iNOS)和內(nèi)皮型一氧化氮合酶(endothelial nitric oxide synthase ,eNOS)蛋白表達的動態(tài)觀察結(jié)果,在HPS形成過程中,實驗第4周肺部iNOS和eNOS的蛋白表達顯著增高,在6周和8周時間點呈明顯的持續(xù)上升趨勢; 2.2肺組織iNOS和eNOS基因表達的動態(tài)觀察結(jié)果,在HPS形成過程中肺部iNOS的基因表達逐漸增加,4周末即有升高,6周末繼續(xù)升高,到8周末達到高峰;eNOS的基因表達4周起也有增高,到8周達到高峰,兩者表達具有相似的規(guī)律。 3.TNF-α和ET-1對HPS形成過程中肺部iNOS和eNOS表達影響的結(jié)果 TNF-α4周時顯著升高(與對照組比較p0.05),6周與4周比有所下降,但仍顯著高于正常對照組,8周升高并達到最高峰;ET-1 4周顯著下降(p0.05),6周與4周比有所升高,但仍低于對照組,8周繼續(xù)升高達到最高峰。TNF-α水平與iNOS、eNOS蛋白和基因之間具有顯著相關(guān)性。 結(jié)論:1. HPS形成過程中,肝臟功能改變是肺功能變化的基礎(chǔ); 2. HPS形成過程中伴隨發(fā)生的IETM,可能是肝功能障礙導致HPS自發(fā)形成的關(guān)鍵; 3. HPS形成過程中,IETM可能對肺部eNOS和iNOS的表達發(fā)揮重要的調(diào)節(jié)作用; 4. IETM對肺部eNOS和iNOS在基因和蛋白水平進行的調(diào)節(jié),可能與TNF-α和ET-1相關(guān)。
[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-偽 and ET-1 on the expression of iNOS and eNOS in lung during HPS formation. The expression of TNF-偽 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-偽. 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- 偽 and ET-1.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R363
本文編號：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-偽 and ET-1 on the expression of iNOS and eNOS in lung during HPS formation. The expression of TNF-偽 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-偽. 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- 偽 and ET-1.
【學位授予單位】：山西醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R363
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