本文選題：電刺激 + 靶基因預(yù)測��； 參考：《廣西醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的：研究預(yù)電刺激小腦頂核誘導(dǎo)大鼠腦缺血再灌注損傷過程中miR-29C對Bak1的調(diào)控作用，，并探討預(yù)電刺激小腦頂核的保護機制。 方法：60只體重280-300g的SD成年雄性大鼠，隨機分成3組：(1)預(yù)電刺激小腦頂核+大腦中動脈阻塞模型（預(yù)電刺激組）；（2）大腦中動脈阻塞模型組（單純造模組）；(3)假手術(shù)組。預(yù)電刺激組的處理為電刺激左側(cè)小腦頂核后制作大腦中動脈阻塞模型，缺血2小時后再灌注，再灌注24小時后處死。通過TTC染色測定梗死體積；動物行為學(xué)評分評估神經(jīng)功能缺損情況；TUNEL組織化學(xué)染色檢測凋亡指數(shù)；芯片分析預(yù)電刺激組和單純造模組間差異表達的miRNA并利用定量PCR驗證；并通過生物信息學(xué)軟件預(yù)測該的miRNA靶基因；最后利用雙熒光報告驗證其與靶基因的關(guān)系。 結(jié)果：預(yù)電刺激組和單純造模組相比，腦梗死體積明顯減少（37.0±8.9%vs62.0±10%）（P0.01）；TUNEL染色檢測的凋亡指數(shù)下降（23.4±2.4%vs58.9±4.0%）(P0.01)；動物行為學(xué)評分降低（1.5±0.52vs2.4±0.51）(P0.01)；芯片分析發(fā)現(xiàn)miR-29C在兩組間表達差異最大； PCR驗證miR-29C在預(yù)電刺激組較單純造模組表達下調(diào)(P0.01)；生物信息學(xué)軟件預(yù)測是Bak1是miR-29C的靶基因；雙熒光報告體外驗證Bak1-wt/rnomiR-29C組的相對熒光強度在Bak1-wt/NC、Bak1-wt/rno-miR-29C、Bak1-Mut/NC和Bak1-Mut/rno-miR-29C4組中最小(P0.01)，并且其余三組間無明顯差異(P0.05) 結(jié)論：預(yù)電刺激大鼠小腦頂核可以減輕腦缺血再灌注損傷；預(yù)電刺激小腦頂核可以下調(diào)miR-29C的表達；Bak1是miR-29C的靶基因；miR-29C是否通過調(diào)控Bak1在預(yù)電刺激誘導(dǎo)的缺血腦保護機制中起作用還需要進一步體內(nèi)驗證。
[Abstract]:Aim: to investigate the regulatory effect of miR-29C on Bak1 during cerebellar fastigial nucleus stimulation in rats with cerebral ischemia-reperfusion injury, and to explore the protective mechanism of preelectric stimulation of cerebellar parietal nucleus. Methods 60 adult Sprague-Dawley male rats weighing 280-300g were randomly divided into three groups: (1) the model of middle cerebral artery occlusion in cerebellar parietal nucleus (); (_ 2) and the sham-operation group (); (_ 3) in prestimulation group. The model of middle cerebral artery occlusion was made after electrical stimulation of left cerebellar fastigial nucleus in preelectric stimulation group. After 2 hours of ischemia and reperfusion, the rats were killed after 24 hours of reperfusion. The infarct volume was measured by TTC staining, the neurological impairment was evaluated by animal behavior score and apoptosis index was detected by Tunel histochemical staining, miRNA differentially expressed between preelectric stimulation group and simple model group was analyzed by microarray and verified by quantitative PCR. The miRNA target gene was predicted by bioinformatics software, and the relationship between miRNA target gene and the target gene was verified by double fluorescence report. Results: compared with the control group, the volume of cerebral infarction was significantly decreased (37.0 鹵8.9%vs62.0 鹵10%) (P0.01), the apoptotic index was decreased (23.4 鹵2.4%vs58.9 鹵4.0%), the animal behavior score was decreased (1.5 鹵0.52vs2.4 鹵0.51) (P0.01), and the expression of miR-29C was found to be the most significant difference between the two groups by microarray analysis. The expression of miR-29C was down-regulated in preelectric stimulation group (P0.01), and the bioinformatics software predicted that Bak1 was the target gene of miR-29C. The double-fluorescence report confirmed in vitro that the relative fluorescence intensity of Bak1-wt / rnomiR-29C group was the smallest in Bak1-wtrno-miR-29C group (P0.01), and the other three groups had no significant difference (P0.05). Conclusion: preelectric stimulation of cerebellar fastigial nucleus can reduce cerebral ischemia-reperfusion injury in rats. The expression of miR-29C can be down-regulated by prestimulation of cerebellar fastigial nucleus. Whether miR-29C, the target gene of miR-29C, may play a role in the protective mechanism of ischemic brain induced by preelectric stimulation needs to be further verified in vivo.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R743.3

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相關(guān)期刊論文 前2條

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