本文選題：斑馬魚(yú)　切入點(diǎn)：橙皮苷　出處：《南方醫(yī)科大學(xué)》2017年碩士論文　論文類(lèi)型：學(xué)位論文

【摘要】：研究背景及目的酒精性脂肪肝(alcoholic fatty liver disease,AFLD)以長(zhǎng)期大量飲酒而導(dǎo)致脂肪在肝細(xì)胞中過(guò)度貯積為主要特征,是酒精性肝病(alcoholic liver disease,ALD)的早期階段,及時(shí)治療可以得到有效控制甚至逆轉(zhuǎn),對(duì)ALD的防治意義重大。中醫(yī)藥治療AFLD歷史悠久且療效顯著,深入研究中藥及其成分對(duì)AFLD的作用效果及機(jī)制,對(duì)AFLD的防治意義重大。橙皮苷是中藥陳皮、枳實(shí)的主要成分,具有抗炎、抗氧化、降膽固醇等多種活性作用,對(duì)AFLD有一定的療效,而具體的作用機(jī)制尚不明確。斑馬魚(yú)因其獨(dú)特的生物學(xué)優(yōu)勢(shì),現(xiàn)已廣泛應(yīng)用于疾病模型的建立和藥物篩選,其中斑馬魚(yú)急性酒精性脂肪肝模型的建立,為進(jìn)一步研究AFLD的發(fā)病機(jī)制及開(kāi)展藥物研究開(kāi)辟了新的途徑�；谝陨媳尘�,本研究利用斑馬魚(yú)急性酒精性脂肪肝模型,旨在進(jìn)一步研究橙皮苷對(duì)AFLD的保護(hù)作用及其機(jī)制,為中藥在AFLD上的應(yīng)用提供依據(jù)。研究方法1.實(shí)驗(yàn)動(dòng)物AB野生型斑馬魚(yú)、肝臟熒光轉(zhuǎn)基因斑馬魚(yú)Tg(1fabp1Oα:eGFP)。2.酒精和藥物處理隨機(jī)挑選4dpf(days post ferti1ization)的斑馬魚(yú)于培養(yǎng)皿中,對(duì)照組只加培養(yǎng)用水,模型組中加入含350mM乙醇的培養(yǎng)用水,培育32小時(shí),建立急性酒精性脂肪肝模型,收集一部分樣品進(jìn)行后續(xù)研究。隨后,隨機(jī)將模型組分為5組:0.1%DMSO 組、模型組、3 個(gè)濃度橙皮苷處理組(25ug/mL、12.5ug/mL、6.25ug/mL),與對(duì)照組一起置于28.5℃恒溫箱培養(yǎng)中培養(yǎng)48小時(shí),收集各組幼魚(yú)進(jìn)行下一步研究。3.試驗(yàn)技術(shù)通過(guò)油紅O染色、尼羅紅染色、HE染色技術(shù)觀察斑馬魚(yú)肝臟形態(tài)學(xué)變化、脂滴沉積情況及組織病理學(xué)改變;運(yùn)用qPCR技術(shù)來(lái)檢測(cè)目標(biāo)基因的mRNA表達(dá)。4.數(shù)據(jù)統(tǒng)計(jì):采用spss20軟件進(jìn)行數(shù)據(jù)分析,計(jì)量資料以均數(shù)±標(biāo)準(zhǔn)差(x±s)表示,P0.05視為差異有統(tǒng)計(jì)學(xué)意義。實(shí)驗(yàn)結(jié)果1.相比對(duì)照組,置于含350mM乙醇的水中培育32小時(shí)的斑馬魚(yú)肝組織形態(tài)改變和脂肪變程度更明顯,證實(shí)成功建立斑馬魚(yú)急性酒精性脂肪肝模型。2.油紅O染色、尼羅紅染色顯示橙皮苷可顯著減輕酒精導(dǎo)致的肝內(nèi)脂滴沉積和肝臟形態(tài)學(xué)改變,斑馬魚(yú)石蠟切片的HE染色證實(shí)橙皮苷能改善酒精導(dǎo)致的肝臟病理變化。說(shuō)明橙皮苷能改善酒精所致的斑馬魚(yú)肝臟脂肪變。3.橙皮苷通過(guò)調(diào)節(jié)酒精代謝、脂質(zhì)代謝以及抵抗內(nèi)質(zhì)網(wǎng)應(yīng)激和DNA損傷來(lái)抵御酒精對(duì)肝臟的損害,具體表現(xiàn)為下調(diào)cyp2y3,cyp3a65,fads2,fasn,hmgcra,hmgcrb,chop,gadd45αa,edem1基因的mRNA表達(dá)量。
[Abstract]:Background and objective\\\. It is of great significance to prevent and cure ALD. Chinese medicine has a long history and remarkable curative effect in the treatment of AFLD. It is very important to study the effect and mechanism of Chinese medicine and its components on AFLD. Hesperidin is the main component of Fructus Aurantii and Fructus Aurantii. Zebrafish have been widely used in the establishment of disease models and drug screening because of their unique biological advantages. The establishment of acute alcoholic fatty liver model of zebrafish opens up a new way to further study the pathogenesis of AFLD and the development of drug research. Based on the above background, the model of acute alcoholic fatty liver of zebrafish was used in this study. The aim of this study was to further study the protective effect of hesperidin on AFLD and its mechanism, and to provide the basis for the application of traditional Chinese medicine in AFLD. Liver fluorescence transgenic zebrafish Tg(1fabp1O 偽: eGFP.2.Alcohol and drug treatment were randomly selected from zebrafish (4dpffdays post fertigation) in petri dish. The control group was treated with culture water only, and the model group was treated with 350mM ethanol for 32 hours. An acute alcoholic fatty liver model was established and some samples were collected for follow-up study. Then the model group was randomly divided into 5 groups: 0.1 DMSO group. The model group with 3 concentrations of hesperidin was cultured in 28.5 鈩，

本文編號(hào)：1569226