發(fā)布時(shí)間：2018-05-30 02:45

  本文選題：自噬 + 環(huán)丙沙星�。� 參考：《成都學(xué)院》2017年碩士論文

【摘要】：背景:研究發(fā)現(xiàn)自噬在治療多種疾病中起著重要的作用,環(huán)丙沙星是親溶酶體抗生素,溶酶體是自噬過程中的關(guān)鍵細(xì)胞器,環(huán)丙沙星可能通過干擾細(xì)胞自噬活性而影響肝細(xì)胞的生理或病理功能。目的:檢測環(huán)丙沙星自噬活性,篩選出環(huán)丙沙星給藥小鼠原代肝細(xì)胞的差異表達(dá)基因,富集環(huán)丙沙星小鼠功能基因以及信號通路,為進(jìn)一步揭示環(huán)丙沙星給藥后的體內(nèi)代謝及分子機(jī)制研究提供理論依據(jù)。方法:17.0±2.0g昆明小鼠,飼養(yǎng)于SPF系統(tǒng)中,按照隨機(jī)原則,分為實(shí)驗(yàn)組和對照組,實(shí)驗(yàn)組小鼠采用灌胃方式給環(huán)丙沙星,對照組予生理鹽水,持續(xù)7天,于第7天通過膠原酶灌注法分離小鼠原代肝細(xì)胞,貼壁后換液,自噬溶酶體熒光探針試劑盒按照比例加入,拍攝熒光圖片,Agilent小鼠全基因組表達(dá)譜芯片檢測基因表達(dá)。結(jié)果:實(shí)驗(yàn)組及對照組都可見自噬熒光亮點(diǎn),但實(shí)驗(yàn)組亮點(diǎn)較小且數(shù)量更少。按照Foldchange≥1.5或≤0.75,組間進(jìn)行t-test分析,檢驗(yàn)標(biāo)準(zhǔn)P0.05,篩選出實(shí)驗(yàn)組與對照組之間差異表達(dá)的基因458個(gè),其中差異表達(dá)上調(diào)基因共有133個(gè),差異表達(dá)下調(diào)基因有325個(gè),并在GENECARDS輸入關(guān)鍵詞autophagy獲得1971個(gè)與自噬相關(guān)的基因,將篩選的差異基因與1971個(gè)自噬相關(guān)的基因進(jìn)行比對,本實(shí)驗(yàn)差異表達(dá)基因中自噬相關(guān)基因表達(dá)上調(diào)9個(gè),而自噬相關(guān)基因表達(dá)下調(diào)27個(gè),KEGG分析結(jié)果顯示16個(gè)顯著通路Pathway,包括:錯(cuò)配修復(fù)基因(Mismatch repair),粘多糖的降解(Glycosaminoglycan degradation),DNA復(fù)制(DNA replication),粘蛋白型O-聚糖的生物合成(Mucin type O-glycan biosynthesis),半乳糖代謝(Galactose metabolism),核苷酸切除修復(fù)(Nucleotide excision repair),同源重組(Homologous recombination),神經(jīng)膠質(zhì)瘤(Glioma),p53信號通路(p53 signaling pathway),鞘脂代謝(Sphingolipid metabolism),磷酸肌醇代謝(Inositol phosphate metabolism),肥厚性心肌病(HCM),蛋白質(zhì)消化吸收(Protein digestion and absorption)。結(jié)論:小鼠原代肝細(xì)胞自噬活性被環(huán)丙沙星抑制,通過生物信息學(xué)分析發(fā)現(xiàn)環(huán)丙沙星可以顯著影響小鼠肝臟基因表達(dá),差異基因富集主要集中在信號傳導(dǎo)、糖代謝等。根據(jù)實(shí)驗(yàn)獲得的信號通路,我們將在后期實(shí)驗(yàn),進(jìn)一步驗(yàn)證環(huán)丙沙星影響小鼠原代肝細(xì)胞自噬活性的主要信號通路以及證實(shí)差異基因之間的相互作用。
[Abstract]:Background: autophagy plays an important role in the treatment of many diseases. Ciprofloxacin is a lysophile antibiotic and lysosome is the key organelle in autophagy. Ciprofloxacin may affect the physiological or pathological function of hepatocytes by interfering with autophagy. Objective: to detect the autophagy activity of ciprofloxacin and to screen the differentially expressed genes in primary liver cells of ciprofloxacin treated mice, and to enrich the functional genes and signal pathways of ciprofloxacin mice. It provides theoretical basis for further study on metabolism and molecular mechanism of ciprofloxacin after administration of ciprofloxacin. Methods the mice were divided into experimental group and control group according to the random principle. The mice in the experimental group were given ciprofloxacin by gavage, and the control group was given normal saline for 7 days. On the 7th day, primary mouse hepatocytes were isolated by collagenase perfusion method, and then injected into the lysosomal fluorescence probe kit. The whole genome expression profile of Agilent mice was detected by fluorescence microarray. Results: both the experimental group and the control group showed fluorescent bright spots of autophagy, but the light spots of the experimental group were smaller and fewer. According to Foldchange 鈮，

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