發(fā)布時(shí)間：2018-05-27 20:22

  本文選題：肝纖維化 + Notch信號(hào)通路��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:肝纖維化是一種慢性肝損傷時(shí)常見(jiàn)的創(chuàng)傷修復(fù)反應(yīng),可由多種病因引起包括酒精或藥物毒性、持續(xù)性病毒感染、遺傳因素等。其核心環(huán)節(jié)是肝星狀細(xì)胞(hepatic stellate cells,HSCs)活化,表達(dá)α-平滑肌肌動(dòng)蛋白(α-SMA),并分泌包括Ⅰ型膠原蛋白(CollagenⅠ,COL1)等多種成分在內(nèi)的細(xì)胞外基質(zhì)(extracellular matrix,ECM)。Notch和Wnt信號(hào)通路普遍分布于生物體內(nèi),精準(zhǔn)調(diào)控著細(xì)胞各項(xiàng)生理進(jìn)程,如生長(zhǎng)、分化、凋亡等。研究顯示這兩條通路可能與肝纖維化的形成關(guān)系密切,本實(shí)驗(yàn)通過(guò)構(gòu)建肝纖維化模型,檢測(cè)模型中Notch和Wnt信號(hào)通路關(guān)鍵因子mRNA的表達(dá)來(lái)探討上述通路對(duì)肝纖維化的影響。方法:1.構(gòu)建模型:選取健康雄性Sprague Dawley大鼠20只,在實(shí)驗(yàn)前禁飲食,隨機(jī)分為正常對(duì)照組(n=10)和肝纖維化模型組(n=10)。每3天1次,于模型組大鼠皮下注入40%四氯化碳(0.6 mL/100 g體重,首劑加倍),正常對(duì)照組則以相同方式注入等量0.9%NaCl,持續(xù)至第8周末,處死所有大鼠,取肝臟組織立即置于低溫環(huán)境冷藏。2.檢測(cè)指標(biāo):提取兩組肝臟標(biāo)本RNA,經(jīng)逆轉(zhuǎn)錄反應(yīng)合成cDNA,采用Q-PCR方法檢測(cè)正常對(duì)照組及肝纖維化模型組α-SMA、COL1α2、Notch3、Wnt5a、β-catenin及GAPDH mRNA表達(dá)量。3.統(tǒng)計(jì)分析:比較Notch與Wnt信號(hào)分子在兩組間表達(dá)的差異性,分析肝纖維化指標(biāo)與Notch、Wnt信號(hào)通路分子相關(guān)性。應(yīng)用SPSS22.0軟件分析所有實(shí)驗(yàn)數(shù)據(jù),其中計(jì)量資料的結(jié)果用均數(shù)±標(biāo)準(zhǔn)差(`x±S)表示,組間均數(shù)之間的比較使用兩獨(dú)立樣本t檢驗(yàn),相關(guān)性分析使用Persion相關(guān)分析法。以P0.05為差異有統(tǒng)計(jì)學(xué)意義。結(jié)果:大鼠肝臟組織中肝纖維化指標(biāo)及Notch、Wnt信號(hào)分子mRNA表達(dá)變化情況:Q-PCR檢測(cè)結(jié)果顯示,與正常對(duì)照組相比,α-SMA、COL1α2、Notch3、Wnt5a表達(dá)增高,差異有統(tǒng)計(jì)學(xué)意義(P0.05),β-catenin表達(dá)無(wú)明顯差異(P0.05)。各項(xiàng)指標(biāo)中,α-SMA與Wnt5a呈正相關(guān)(r=0.689,P0.05),COL1α2與Notch3呈正相關(guān)(r=0.676,P0.05)。結(jié)論:Notch及Wnt非經(jīng)典信號(hào)通路參與了肝纖維化的發(fā)生、發(fā)展過(guò)程。選擇性干預(yù)Notch3、Wnt5a轉(zhuǎn)導(dǎo)可能成為抗肝纖維化新途徑。
[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 偽 -smooth muscle actin (偽 -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 偽 -SMA-COL1 偽 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 鹵standard deviation (`x 鹵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 偽 -SMA-COL1 偽 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 偽 -SMA and Wnt5a. The correlation between 偽 -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.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：R575.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 Kai Zhang;Yan-Qiong Zhang;Wen-Bing Ai;Qing-Ting Hu;Qiao-Juan Zhang;Lin-Yan Wan;Xiao-Lian Wang;Chang-Bai Liu;Jiang-Feng Wu;;Hes1,an important gene for activation of hepatic stellate cells,is regulated by Notch1 and TGF-β/BMP signaling[J];World Journal of Gastroenterology;2015年03期

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本文編號(hào)：1943618