本文選題：長鏈非編碼RNAs　切入點(diǎn)：骨橋蛋白　出處：《第二軍醫(yī)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：【目的】運(yùn)用生物信息學(xué)方法檢測與骨橋蛋白相關(guān)的Lnc RNAs在草酸鈉誘導(dǎo)結(jié)晶腎損傷中的差異變化,初步探討骨橋蛋白相關(guān)Lnc RNAs在結(jié)晶腎損傷中的作用機(jī)制。【方法】利用草酸鈉刺激腎小管上皮細(xì)胞(HK-2)構(gòu)建結(jié)晶腎損傷模型。采用實(shí)時定量PCR(RT-PCR)技術(shù),在通過草酸鈉刺激腎小管上皮細(xì)胞(HK-2)構(gòu)建的細(xì)胞模型中驗(yàn)證骨橋蛋白表達(dá)水平。運(yùn)用RNA干擾技術(shù)構(gòu)建骨橋蛋白敲低細(xì)胞模型。選用Arraystar公司提供的人類全基因組lnc RNA芯片(V4.0)對3組骨橋蛋白干擾組和3組對照組腎小管上皮細(xì)胞結(jié)晶模型進(jìn)行骨橋蛋白(OPN)相關(guān)lnc RNAs以及m RNAs差異基因表達(dá)水平檢測。其中l(wèi)nc RNAs來源于權(quán)威公共轉(zhuǎn)錄組數(shù)據(jù)庫(包括Refseq、UCSC knowngenes、Gencode等)。差異基因篩選條件是差異倍數(shù)1.5,P0.05。對差異基因進(jìn)行基因本體論(GO)及信號通路分析(KEGG Pathway)等生物信息學(xué)分析�！窘Y(jié)果】1.通過草酸鈉刺激腎小管上皮細(xì)胞成功構(gòu)建結(jié)晶腎損傷細(xì)胞模型以及通過RNA干擾技術(shù)構(gòu)建骨橋蛋白干擾模型。2.全基因組lnc RNA芯片提示在腎小管上皮細(xì)胞結(jié)晶腎損傷模型中存在骨橋蛋白相關(guān)差異表達(dá)的lnc RNAs,共有583個lnc RNAs存在差異表達(dá),其中骨橋蛋白干擾組表達(dá)上調(diào)354個,下調(diào)229個;共有235個m RNAs存在差異表達(dá),其中干擾組上調(diào)139個,下調(diào)96個。3.通過基因本體論(GO)及信號通路分析(KEGG Pathway)等生物信息學(xué)分析表明,差異表達(dá)的lnc RNAs與細(xì)胞生長、酶活性調(diào)節(jié)、PI3k/Akt/NF-κB信號通路、Wnt信號通路等相關(guān)聯(lián)。KEGG信號通路分析結(jié)果提示,上調(diào)lnc RNAs與14條通路相關(guān),而下調(diào)lnc RNAs與5條通路相關(guān)。綜合分析提示NOD樣受體信號通路、PI3k/Akt/NF-κB信號通路、Wnt信號通路等炎癥相關(guān)通路顯著富集。【結(jié)論】體外建立人腎小管上皮細(xì)胞結(jié)晶腎損傷模型中存在骨橋蛋白相關(guān)差異Lnc RNAs表達(dá)。骨橋蛋白相關(guān)lnc RNAs參與草酸鹽結(jié)晶腎損傷發(fā)病機(jī)制,未來可作為候選生物標(biāo)記物為臨床上結(jié)晶所致腎損傷提供理論及實(shí)驗(yàn)依據(jù)。
[Abstract]:[objective] to detect the differential changes of Lnc RNAs associated with osteopontin in sodium oxalate induced renal injury by bioinformatics. To explore the mechanism of osteopontin associated Lnc RNAs in renal injury. [methods] the model of renal injury was established by sodium oxalate stimulation of renal tubular epithelial cells (HK-2). The expression level of osteopontin was verified in a cell model constructed by sodium oxalate stimulation of renal tubular epithelial cells (HK-2). Osteopontin knockdown cell model was constructed by using RNA interference technique. The whole human genome lnc provided by Arraystar was selected. RNA microarray V4.0) was used to detect the differential gene expression levels of lnc RNAs and m RNAs related to osteopontin interference group and renal tubular epithelial cell crystallization model of 3 groups and 3 control groups. Lnc RNAs originated from authoritative common transcription. Group database (including RefseqUCSC knowngenesl Gencode et al. The screening condition of differential gene is 1.5mv P0.05.The differential gene is analyzed by gene ontology and signal pathway analysis KEGG Pathway. [results] 1. Stimulating renal tubules by sodium oxalate. The successful construction of crystalline renal injury cell model by skin cells and the construction of osteopontin interference model by RNA interference technology. 2. The whole genome lnc RNA chip indicates that there is osteopontin correlation in renal tubular epithelial cell crystal kidney injury model. Among the differentially expressed lnc RNASs, 583 lnc RNAs were differentially expressed. There were 354 up-regulated and 229 down-regulated osteopontin interference groups, and 235 m RNAs were differentially expressed, in which 139 m RNAs were up-regulated and 96 路3.The results of bioinformatics analysis, such as gene ontology and signal pathway analysis, showed that, The differential expression of lnc RNAs was associated with cell growth, enzyme activity regulation, PI3k / NF- 魏 B signaling pathway and Wnt signaling pathway. The results showed that the up-regulation of lnc RNAs was associated with 14 pathways. However, down-regulation of lnc RNAs was associated with 5 pathways. Comprehensive analysis showed that the PI3k / Akt / NF- 魏 B signaling pathway and Wnt signaling pathway were significantly enriched. [conclusion] the model of renal injury induced by crystallization of human renal tubular epithelial cells was established in vitro. Osteopontin related lnc RNAs was involved in the pathogenesis of oxalate crystal kidney injury. It can be used as a candidate biomarker to provide theoretical and experimental evidence for renal injury induced by clinical crystallization.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R692

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相關(guān)期刊論文 前1條

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本文編號：1650832