背景:腦卒中是嚴(yán)重威脅著人類的生命健康的重大疾病之一。Nurr1是核受體4家族的一個(gè)成員。Nurr1在炎癥反應(yīng)中明顯下調(diào)且參與帕金森疾病中神經(jīng)元的死亡。microRNAs在腦缺血再灌注損傷中發(fā)揮重要作用參與多種關(guān)鍵因子的轉(zhuǎn)錄水平的調(diào)節(jié)。Nurr1的異常表達(dá)與多種信號通路相關(guān)有,但miRNA與Nurr1的調(diào)節(jié)關(guān)系在腦缺血再灌注損傷中幾乎未見報(bào)道。目的:本研究目的在于:（1）在體內(nèi)和體外實(shí)驗(yàn)探索Nurr1在腦缺血再灌注損傷中的表達(dá)情況及Nurr1相關(guān)的microRNAs;（2）在體內(nèi)和體外實(shí)驗(yàn)探索Nurr1蛋白的轉(zhuǎn)錄后調(diào)控網(wǎng)絡(luò),及其在活化的膠質(zhì)細(xì)胞的具體作用機(jī)制;（3）探究miR-145-5p/Nurr1/TNF-a軸信號在大鼠腦缺血再灌注損傷中的具體功能。方法:（1）原代神經(jīng)元細(xì)胞和小膠質(zhì)細(xì)胞培養(yǎng)。（2）神經(jīng)元細(xì)胞與小膠質(zhì)細(xì)胞共培養(yǎng)模型建立及OGD/R實(shí)驗(yàn)。（3）大鼠大腦中動脈梗塞再灌注（MCAO/R）模型構(gòu)建,神經(jīng)功能學(xué)結(jié)果與梗塞體積分析。（4）RNA提取,逆轉(zhuǎn)錄和實(shí)時(shí)定量qPCR。（5）大鼠側(cè)腦室注射miR-145-5p mimic和miR-145-5p antagomir。（6）N... 

【文章來源】：重慶醫(yī)科大學(xué)重慶市

【文章頁數(shù)】：81 頁

【學(xué)位級別】：博士

【部分圖文】：

MCAO模型后，TTC染色

(8)檢測 Nurr1 3’UTR 片段與 miRNA 結(jié)合情況的對照實(shí)驗(yàn)分組：第一組 pMIR-質(zhì)粒不加 Nurr1 3’UTR 片段 + miRNA圖 2 HEK293T 細(xì)胞中 miR145-5p 與 Nurr1 3’UTR 片段結(jié)合的預(yù)實(shí)

47圖 4 直接靶向 Nurr1 3′ UTR 的 miRNAs 篩選Figure 4 Screening for miRNAs that directly target the 3′UTR of Nurr1.(A) The infarct volume was negatively correlated with Nurr1 expression some extent byPearson correlation test, but was not statistically significant (correlation coefficient -0.457, P =0.362). (B) Heat map of predicted miRNAs expression in cerebral cortex tissue from sham andMCAO/R animals (one-way ANOVA, p < 0.05). (C) The predicted binding sites of selected miRNA(in green color) to the 3'UTR of Nurr1 (in red color) is mapped in this figure. Nucleotides whichwere altered for mutational studies are marked in gray color of background. (D) Quantitation of th

【參考文獻(xiàn)】：
期刊論文
[1]Novel splicing variant of the human orphan nuclear receptor Nurr1 gene[J]. 徐評議,樂衛(wèi)東.  Chinese Medical Journal. 2004(06)



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