Notch及Wnt信号通路与肝纤维化指标相关研究

发布时间：2018-05-27 20:22

本文选题：肝纤维化 + Notch信号通路　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:肝纤维化是一种慢性肝损伤时常见的创伤修复反应,可由多种病因引起包括酒精或药物毒性、持续性病毒感染、遗传因素等。其核心环节是肝星状细胞(hepatic stellate cells,HSCs)活化,表达α-平滑肌肌动蛋白(α-SMA),并分泌包括Ⅰ型胶原蛋白(CollagenⅠ,COL1)等多种成分在内的细胞外基质(extracellular matrix,ECM)。Notch和Wnt信号通路普遍分布于生物体内,精准调控着细胞各项生理进程,如生长、分化、凋亡等。研究显示这两条通路可能与肝纤维化的形成关系密切,本实验通过构建肝纤维化模型,检测模型中Notch和Wnt信号通路关键因子mRNA的表达来探讨上述通路对肝纤维化的影响。方法:1.构建模型:选取健康雄性Sprague Dawley大鼠20只,在实验前禁饮食,随机分为正常对照组(n=10)和肝纤维化模型组(n=10)。每3天1次,于模型组大鼠皮下注入40%四氯化碳(0.6 mL/100 g体重,首剂加倍),正常对照组则以相同方式注入等量0.9%NaCl,持续至第8周末,处死所有大鼠,取肝脏组织立即置于低温环境冷藏。2.检测指标:提取两组肝脏标本RNA,经逆转录反应合成cDNA,采用Q-PCR方法检测正常对照组及肝纤维化模型组α-SMA、COL1α2、Notch3、Wnt5a、β-catenin及GAPDH mRNA表达量。3.统计分析:比较Notch与Wnt信号分子在两组间表达的差异性,分析肝纤维化指标与Notch、Wnt信号通路分子相关性。应用SPSS22.0软件分析所有实验数据,其中计量资料的结果用均数±标准差(`x±S)表示,组间均数之间的比较使用两独立样本t检验,相关性分析使用Persion相关分析法。以P0.05为差异有统计学意义。结果:大鼠肝脏组织中肝纤维化指标及Notch、Wnt信号分子mRNA表达变化情况:Q-PCR检测结果显示,与正常对照组相比,α-SMA、COL1α2、Notch3、Wnt5a表达增高,差异有统计学意义(P0.05),β-catenin表达无明显差异(P0.05)。各项指标中,α-SMA与Wnt5a呈正相关(r=0.689,P0.05),COL1α2与Notch3呈正相关(r=0.676,P0.05)。结论:Notch及Wnt非经典信号通路参与了肝纤维化的发生、发展过程。选择性干预Notch3、Wnt5a转导可能成为抗肝纤维化新途径。
[Abstract]:Objective: hepatic fibrosis is a common wound repair reaction in chronic liver injury. It can be caused by many causes, including alcohol or drug toxicity, persistent virus infection, genetic factors and so on. Its core link is the activation of hepatic stellate cells (HSCs), the expression of 伪 -smooth muscle actin (伪 -SMAN), and the secretion of extracellular matrix (extracellular matrix), such as extracellular matrix, ECM, Notch and Wnt signaling pathway. Precise regulation of cell physiological processes, such as growth, differentiation, apoptosis and so on. The study showed that these two pathways may be closely related to the formation of hepatic fibrosis. In order to investigate the effects of these pathways on hepatic fibrosis, we constructed a model of liver fibrosis and detected the expression of mRNA, a key factor of Notch and Wnt signaling pathway. Method 1: 1. Model: 20 healthy male Sprague Dawley rats were randomly divided into normal control group (n = 10) and liver fibrosis model group (n = 10). Once every 3 days, the rats in the model group were subcutaneously injected with 40% carbon tetrachloride 0.6 mL/100 g body weight, the first dose was doubled, while the normal control group was injected with the same amount of 0.9% NaCl1 in the same way until the 8th week, and all the rats were killed. The liver tissue was immediately stored in hypothermia. Q-PCR method was used to detect the expression of 伪 -SMA-COL1 伪 2Notch3Nt5a, 尾 -catenin and GAPDH mRNA in normal control group and liver fibrosis model group. Statistical analysis: to compare the difference of expression of Notch and Wnt signal molecules between the two groups, and to analyze the correlation between hepatic fibrosis index and Notchnt-Wnt signal pathway molecules. All the experimental data were analyzed by SPSS22.0 software. The results of measurement data were expressed as mean 卤standard deviation (`x 卤S). Two independent samples t test were used to compare the mean between groups, and Persion correlation analysis was used to analyze the correlation. P0.05 as the difference was statistically significant. Results: compared with the normal control group, the expression of 伪 -SMA-COL1 伪 2Notch3nt5a in liver fibrosis index and the expression of Notch3Notch3Wnt5a in rat liver tissue were higher than those in normal control group (P 0.05), but there was no significant difference in 尾 -catenin expression (P0.05). There was a positive correlation between 伪 -SMA and Wnt5a. The correlation between 伪 -SMA and Notch3 was 0. 676%, 0. 676% (P 0. 05) and 0. 05% (P 0. 05). Conclusion the non-classical signal pathways of Wnt and Notch are involved in the pathogenesis and development of hepatic fibrosis. Selective intervention of Notch3 and Wnt5a transduction may be a new way of anti-hepatic fibrosis.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R575.2

【参考文献】