MRTF-A調(diào)控miR-1273g-3p對(duì)Aβ 25-35 損傷的SH-SY5Y細(xì)胞自噬的影響及機(jī)制研究
發(fā)布時(shí)間：2021-01-29 11:39
目的:在Aβ25-35誘導(dǎo)人源性SH-SY5Y損傷細(xì)胞上,篩選能差異化表達(dá)并與MRTF-A相關(guān)的miRNA,探討MRTF-A介導(dǎo)miRNA對(duì)Aβ25-35誘導(dǎo)的SH-SY5Y細(xì)胞自噬的影響及機(jī)制。方法:1.在Aβ25-35誘導(dǎo)SH-SY5Y損傷細(xì)胞模型中,用基因芯片篩選與MRTF-A調(diào)控相關(guān)的miRNA:建立細(xì)胞模型;細(xì)胞分組(control,Aβ25-35,MRTF-A,MRTF-A+Aβ25-35)。轉(zhuǎn)染MRTF-A質(zhì)粒48h后,給予40μmol/L Aβ25-35處理,24h后,用western blot方法檢測(cè)MRTF-A的蛋白表達(dá)水平,并提取RNA。用Agilent RNA 6000 Nano/Pico Assay法檢測(cè)各組RNA質(zhì)量合格后,使用miRNA One Array芯片技術(shù)分析各組中發(fā)生顯著差異變化的miRNA。得到各組顯著變化的miRNA,再通過各組間相互比較得到MRTF-A調(diào)控的miRNA。篩選出差異性表達(dá)最為顯著的miRNA,并用...
【文章來源】：武漢科技大學(xué)湖北省
【文章頁(yè)數(shù)】：62 頁(yè)
【學(xué)位級(jí)別】：碩士
【部分圖文】：
SH-SY5Y細(xì)胞模型的建立.轉(zhuǎn)染MRTF-A質(zhì)粒2.5μg至SH-SY5Y細(xì)胞中，48h后，給予40μmol/LAβ25-35處理24h（A）激光共聚焦檢測(cè)MRTF-A質(zhì)粒轉(zhuǎn)染結(jié)果，綠色為GFP
武漢科技大學(xué)碩士學(xué)位論文表 1 各組總 RNA 質(zhì)檢結(jié)果Tab 1 Total RNA content in each group樣本 A260/A280 A260/A230 RNA 總量 RIN 值vector 1.88 2.31 20.50 6.10Vector+ Aβ25-351.87 1.82 12.78 5.80MRTF-A+ Aβ25-351.93 1.45 6.38 6.10MRTF-A 1.86 2.32 24.54 5.70
圖 3. miRNA 基因芯片結(jié)果分析.A：miRNA 基因芯片聚類分析圖。聚類分析圖中每一行為一張芯片，每一列為一個(gè)基因探針。圖中 AM 指 MRTF-A+ Aβ25-35組，M 指 MRTF-A 組，C指 vector 陰性對(duì)照組，A 指 Aβ25-35組。紅綠色階標(biāo)示了探針訊號(hào)上/下調(diào)的變化幅度。線的長(zhǎng)度則代表距離指標(biāo)同簇樣本或基因的相似性高(模式相近)。探針的訊號(hào)最大值與訊號(hào)最小值的差異>160 為有用信號(hào)，篩選出前 295 基因探針進(jìn)入分析。B：venn 圖表示差異 miRNA，藍(lán)色框里顯示差異表達(dá)的miRNA，上面圖表示MRTF-A上調(diào)的miRNA，下面圖表示MRTF-A下調(diào)的 miRNA。Fig 3. miRNA gene chip results. (A) microarray cluster analysis of miRNA. Each row is a chip,and each column is a gene probe. AM refers to MRTF-A+ Aβ25-35group, M refers to MRTF-Agroup, C refers to negative control group, and A refers to Aβ25-35group. The red and green bandsindicate the amplitude of the probe signal up / down changes. The length of the line represents thedistance between the same cluster indicators, the gene or gene similarity (mode similar). Thedifference between the signal maximum value and the signal minimum value of all probes in theprobe is greater than 160, and the first 295 gene probes are screened into the analysis. (B) venn
【參考文獻(xiàn)】：
碩士論文
[1]MRTF-A對(duì)Tg2576小鼠腦神經(jīng)元凋亡和自噬的影響及機(jī)制研究[D]. 肖彎.武漢科技大學(xué) 2016
本文編號(hào)：3006823
【文章來源】：武漢科技大學(xué)湖北省
【文章頁(yè)數(shù)】：62 頁(yè)
【學(xué)位級(jí)別】：碩士
【部分圖文】：
SH-SY5Y細(xì)胞模型的建立.轉(zhuǎn)染MRTF-A質(zhì)粒2.5μg至SH-SY5Y細(xì)胞中，48h后，給予40μmol/LAβ25-35處理24h（A）激光共聚焦檢測(cè)MRTF-A質(zhì)粒轉(zhuǎn)染結(jié)果，綠色為GFP
武漢科技大學(xué)碩士學(xué)位論文表 1 各組總 RNA 質(zhì)檢結(jié)果Tab 1 Total RNA content in each group樣本 A260/A280 A260/A230 RNA 總量 RIN 值vector 1.88 2.31 20.50 6.10Vector+ Aβ25-351.87 1.82 12.78 5.80MRTF-A+ Aβ25-351.93 1.45 6.38 6.10MRTF-A 1.86 2.32 24.54 5.70
圖 3. miRNA 基因芯片結(jié)果分析.A：miRNA 基因芯片聚類分析圖。聚類分析圖中每一行為一張芯片，每一列為一個(gè)基因探針。圖中 AM 指 MRTF-A+ Aβ25-35組，M 指 MRTF-A 組，C指 vector 陰性對(duì)照組，A 指 Aβ25-35組。紅綠色階標(biāo)示了探針訊號(hào)上/下調(diào)的變化幅度。線的長(zhǎng)度則代表距離指標(biāo)同簇樣本或基因的相似性高(模式相近)。探針的訊號(hào)最大值與訊號(hào)最小值的差異>160 為有用信號(hào)，篩選出前 295 基因探針進(jìn)入分析。B：venn 圖表示差異 miRNA，藍(lán)色框里顯示差異表達(dá)的miRNA，上面圖表示MRTF-A上調(diào)的miRNA，下面圖表示MRTF-A下調(diào)的 miRNA。Fig 3. miRNA gene chip results. (A) microarray cluster analysis of miRNA. Each row is a chip,and each column is a gene probe. AM refers to MRTF-A+ Aβ25-35group, M refers to MRTF-Agroup, C refers to negative control group, and A refers to Aβ25-35group. The red and green bandsindicate the amplitude of the probe signal up / down changes. The length of the line represents thedistance between the same cluster indicators, the gene or gene similarity (mode similar). Thedifference between the signal maximum value and the signal minimum value of all probes in theprobe is greater than 160, and the first 295 gene probes are screened into the analysis. (B) venn
【參考文獻(xiàn)】：
碩士論文
[1]MRTF-A對(duì)Tg2576小鼠腦神經(jīng)元凋亡和自噬的影響及機(jī)制研究[D]. 肖彎.武漢科技大學(xué) 2016
本文編號(hào)：3006823
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