脾切除对肝纤维化大鼠肝脏TGF-β1、bFGF和α-SMA表达影响的研究

发布时间：2018-03-14 19:25

本文选题：肝纤维化　切入点：脾切除术　出处：《皖南医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:本研究通过建立肝纤维化大鼠模型,动态观察模型大鼠肝脏组织内TGF-β1、b FGF和α-SMA的表达,探讨TGF-β1、b FGF在肝纤维化进程中的作用与机制;通过对肝纤维化早、中、晚期大鼠行脾切除术,观察肝脏中TGF-β1、b FGF和α-SMA表达的变化,探讨脾脏与肝纤维化发展机制的关系。方法:实验一:按照随机原则,将50只雄性SD大鼠分为正常组、建模2周组、建模4周组、建模6周组和建模8周组,对模型组大鼠腹腔注射40%四氯化碳溶液,剂量为2ml/kg,每周2次,共计8周,建立肝纤维化模型;正常组于建模前处死,其余分别于建模第2、4、6和8周处死。实验二:按照随机原则,将48只雄性SD大鼠分为A、B、C三大组,对模型组大鼠腹腔注射40%四氯化碳溶液,剂量为2ml/kg,每周2次,共计8周,建立肝纤维化模型;每一大组设立实验组和对照组,建模第2周、4周、6周分别对A、B、C三组实验组大鼠行脾切除术,各对照组同期行假手术;建模至第8周处死所有大鼠。留取肝脏标本,HE染色观察肝脏组织的病理变化,免疫组化检测各组大鼠肝脏组织内TGF-β1、b FGF和α-SMA的表达;应用实时荧光定量PCR(q PCR)检测各组大鼠肝脏组织内TGF-β1m RNA、b FGFm RNA和α-SMAm RNA的水平。结果:实验一:(1)免疫组化染色结果显示:随着造模时间的延长,各模型组肝脏组织中TGF-β1、b FGF和α-SMA的表达逐渐增加。模型组肝脏组织中TGF-β1、b FGF和α-SMA的表达均较正常组表达高(P0.05)。TGF-β1、b FGF与α-SMA的表达呈正相关。(2)实时荧光定量PCR检测结果显示:随着造模时间的延长,TGF-β1m RNA、b FGFm RNA和α-SMAm RNA水平逐渐升高。模型组肝脏内TGF-β1m RNA、b FGFm RNA和α-SMAm RNA水平均较正常对照组升高(P0.05)。实验二:(1)免疫组化染色显示各实验组肝脏组织内TGF-β1、b FGF和α-SMA的表达均较同期对照组低(P0.05)。A、B、C三大组实验组作单因素方差分析SNK检验进行均数的两两比较,结果显示肝纤维化早期实施脾切除术,肝脏组织内TGF-β1、b FGF和α-SMA表达最低(P0.05)。(2)实时荧光定量PCR检测显示各实验组肝脏组织内TGF-β1m RNA、b FGFm RNA和α-SMAm RNA的水平均较同期对照组低(P0.05),A、B、C三大组实验组作单因素方差分析SNK检验进行均数的两两比较,结果显示早期肝纤维化实施脾切除术,肝脏组织内TGF-β1m RNA、b FGFm RNA和α-SMAm RNA水平最低(P0.05)。结论:1.随着肝纤维化的发展,大鼠肝脏组织内TGF-β1、b FGF和α-SMA的表达及TGF-β1m RNA、b FGFm RNA和α-SMAm RNA水平逐渐增高。2.脾切除可明显降低肝内TGF-β1、b FGF和α-SMA的表达,延缓肝纤维化的发展。脾脏在肝纤维化发生发展中起重要作用。3.肝纤维化早期行脾切除术较中晚期延缓肝纤维发展效果明显。
[Abstract]:Objective: to investigate the role and mechanism of TGF- 尾 1b FGF and 伪 -SMA in liver fibrosis by establishing a rat model of hepatic fibrosis, and to investigate the mechanism of TGF- 尾 1b FGF in the process of hepatic fibrosis by observing the expression of TGF- 尾 1b FGF and 伪 -SMA in the liver tissue of the model rats, and to explore the role and mechanism of TGF- 尾 1b FGF in the process of liver fibrosis. The expression of TGF- 尾 1b FGF and 伪 -SMA in the liver was observed after splenectomy, and the relationship between the spleen and the development of hepatic fibrosis was investigated. Methods: experiment 1: according to the random principle, 50 male Sprague-Dawley rats were divided into normal group and model group for 2 weeks. The rats in the model group were injected intraperitoneally with 40% carbon tetrachloride solution at a dose of 2 ml / kg twice a week for 8 weeks, and the normal group was killed before modeling, and the rats in the model group were given intraperitoneal injection of 40% carbon tetrachloride solution for 4 weeks, 6 weeks and 8 weeks respectively. The other rats were killed at the 6th and 8th week of the model. Experiment 2: according to the random principle, 48 male SD rats were divided into three groups: the rats in the model group were injected with 40% carbon tetrachloride solution intraperitoneally at the dose of 2 ml / kg twice a week for 8 weeks. Liver fibrosis model was established in each group, each group was divided into experimental group and control group. The rats in group A BX C were treated with splenectomy and sham operation were performed at the second week, 4 weeks and 6 weeks, respectively. From the 8th week, all the rats were killed. The pathological changes of liver tissue were observed by HE staining and the expression of TGF- 尾 1b FGF and 伪 -SMA in the liver tissues of each group were detected by immunohistochemistry. The levels of TGF- 尾 1m RNAb FGFm RNA and 伪 -SMAm RNA in liver tissue of rats in each group were detected by real-time fluorescence quantitative PCR(q. The expression of TGF- 尾 1nb FGF and 伪 -SMA in liver tissue of model group was increased gradually, and the expression of TGF- 尾 1nb FGF and 伪 -SMA in model group was higher than that in normal group. The positive correlation between TGF- 尾 1b FGF and 伪 -SMA expression was observed by real-time quantitative PCR. The levels of TGF- 尾 1m RNAb FGFm RNA and 伪 -SMAm RNA in the liver of the model group were higher than those of the normal control group (P 0.05). The expression of TGF- 尾 1mRNAb FGFm RNA and 伪 -SMAm RNA in the liver of the model group was higher than that of the normal control group (experiment 2: 1) immunohistochemical staining showed that the levels of TGF- 尾 1mRNAb and 伪 -SMAm RNA in the liver tissue of each experimental group were higher than those of the control group. The expression of FGF and 伪 -SMA was lower than that of control group (P 0.05). The SNK test of single factor analysis of variance (SNK) was used to compare the mean number of the three experimental groups with single factor analysis of variance (ANOVA), and the expression of 伪 -SMA was lower than that of the control group. The results showed that splenectomy was performed in the early stage of hepatic fibrosis. Detection of TGF- 尾 1b FGF and 伪 -SMA expression in liver tissue by real-time fluorescence quantitative PCR analysis showed that the levels of TGF- 尾 1m RNAb FGFm RNA and 伪 -SMAm RNA in liver tissue of each experimental group were lower than those of the control group. The results of single factor ANOVA SNK test showed that TGF- 尾 1mRNAb FGFm RNA and 伪 -SMAm RNA were lower than those of the control group. A pairwise comparison of the mean, The results showed that the levels of TGF- 尾 1m RNAb FGFm RNA and 伪 -SMAm RNA were the lowest in liver tissues after splenectomy. Conclusion: 1. The expression of TGF- 尾 1mRNAb FGF and 伪 -SMA and the levels of TGF- 尾 1 RNAb FGFm RNA and 伪 -SMAm RNA increased gradually. 2. Splenectomy significantly decreased the expression of TGF- 尾 1b FGF and 伪 -SMA. Spleen plays an important role in the development of liver fibrosis. 3. Splenectomy in the early stage of liver fibrosis is more effective than that in the middle and late stage of liver fibrosis.
【学位授予单位】：皖南医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R657.3

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