本文選題：MicroRNA-29c + BIRC2�。� 參考：《廣西醫(yī)科大學(xué)》2014年碩士論文

【摘要】：目的探討MicroRNA-29c（miR-29c）在預(yù)電刺激小腦頂核后大鼠局灶性腦缺血再灌注損傷中的保護(hù)機(jī)制。 方法將60只Sprague Dawley雄性大鼠采用隨機(jī)數(shù)字表法分為3組：（1）假手術(shù)組；（2）單純?cè)炷＝M；（3）預(yù)電刺激組：即首先預(yù)電刺激小腦頂核1h，24h后再行右側(cè)局灶性腦缺血，2h后再灌注。然后用TTC染色檢測(cè)腦梗死體積比例；RT-qPCR檢測(cè)miR-29c、BIRC2mRNA表達(dá)量；Western-blotting檢測(cè)BIRC2蛋白表達(dá)量；TUNEL染色檢測(cè)凋亡的細(xì)胞數(shù)；構(gòu)建BIRC2熒光素酶報(bào)告基因載體，雙熒光素酶報(bào)告基因檢測(cè)系統(tǒng)檢測(cè)熒光素酶的表達(dá)變化。 結(jié)果（1）腦梗死體積比例測(cè)定結(jié)果：預(yù)電刺激組與單純?cè)炷＝M相比，腦梗死體積比例減小30.2%（p0.01）。（2）PCR檢測(cè)結(jié)果：假手術(shù)組、預(yù)電刺激組、單純?cè)炷＝M：miR-29c的相對(duì)表達(dá)量分別為（1.0、0.343±0.009、0.955±0.212），預(yù)電刺激組較單純?cè)炷＝M明顯下降，下降約3倍（p0.01）；BIRC2mRNA的相對(duì)表達(dá)量分別為（1.0、2.404±0.330、1.168±0.247），預(yù)電刺激組較單純?cè)炷＝M明顯增高，增高2倍多（p0.01）。（3）BIRC2蛋白表達(dá)量檢測(cè)結(jié)果：假手術(shù)組、預(yù)電刺激組、單純?cè)炷＝MBIRC2蛋白的相對(duì)表達(dá)量分別為（0.5±0.364、1.407±0.270、0.939±0.227），預(yù)電刺激組較單純?cè)炷＝M明顯增加，，約增高1.5倍（p0.05）。（4）TUNEL凋亡細(xì)胞數(shù)檢測(cè)結(jié)果：預(yù)電刺激組凋亡細(xì)胞數(shù)（22.37±1.27）較單純?cè)炷＝M凋亡細(xì)胞數(shù)（35.9±1.93）明顯減少，約減少37.68%（p0.01）。（5）雙熒光素酶報(bào)告基因系統(tǒng)顯示：轉(zhuǎn)染野生型載體組的熒光素酶活性顯著下調(diào)，而轉(zhuǎn)染突變型載體組的熒光素酶活性變化不明顯，說明miR-29c可特異性抑制包含有BIRC23’UTR野生型識(shí)別元件報(bào)告基因的表達(dá)（p0.01）。 結(jié)論預(yù)電刺激小腦頂核對(duì)其后的腦缺血再灌注損傷有保護(hù)作用，其機(jī)制可能與miR-29c通過靶向調(diào)節(jié)BIRC2的表達(dá)、減輕局灶性腦缺血再灌注損傷有關(guān)。
[Abstract]:Objective to investigate the protective mechanism of microRNA-29cmmiR-29c in focal cerebral ischemia-reperfusion injury after prestimulation of cerebellar parietal nucleus in rats.Methods Sixty Sprague Dawley male rats were randomly divided into 3 groups: control group (n = 1) sham operation group (n = 2) model group (n = 2) prestimulation group (n = 3): first prestimulation of cerebellar fastigial nucleus for 24 h and then right focal cerebral ischemia for 2 h and then reperfusion.Then TTC staining was used to detect the proportion of cerebral infarction volume and RT-qPCR was used to detect the expression of miR-29cnBIRC2 mRNA. Western-blotting was used to detect the expression of BIRC2 protein. Tunel staining was used to detect the number of apoptotic cells, and the BIRC2 luciferase reporter gene vector was constructed.The expression of luciferase was detected by double luciferase reporter gene detection system.Results 1) the ratio of cerebral infarction volume was measured: compared with the control group, the ratio of cerebral infarction volume in the prestimulation group was smaller than that in the control group: sham operation group, preelectric stimulation group, sham operation group, preelectric stimulation group,The relative expression of miR-29c in the control group was 0.343 鹵0.009 鹵0.955 鹵0.212g, respectively. The relative expression of BIRC2 mRNA in the prestimulation group was significantly lower than that in the control group, and the relative expression of BIRC2 mRNA in the prestimulation group was about 3 times lower than that in the control group. The relative expression of BIRC2 mRNA in the prestimulation group was significantly higher than that in the control group, and the relative expression of BIRC2 mRNA in the preelectric stimulation group was significantly higher than that in the control group, and the relative expression of miR-29c mRNA in the prestimulation group was significantly higher than that in the control group.The results showed that the relative expression of BIRC2 protein in sham operation group, preelectric stimulation group and simple model group was 0.5 鹵0.364 鹵0.270 鹵0.939 鹵0.227, respectively. The expression of BIRC2 protein in preelectric stimulation group was significantly higher than that in the simple model group, and the expression of BIRC2 protein was significantly higher in the prestimulation group than in the control group, and the relative expression of BIRC2 protein in the sham operation group, preelectric stimulation group and simple model group were 0.5 鹵0.364 鹵0.270 鹵0.939 鹵0.227, respectively.The number of apoptotic cells in preelectric stimulation group (22.37 鹵1.27) was significantly lower than that in model group (35.9 鹵1.93).The system of double luciferase report gene showed that the luciferase activity of the wild-type vector group decreased significantly, but the luciferase activity of the mutant vector group did not change significantly.These results suggest that miR-29c can specifically inhibit the expression of a reporter gene containing BIRC23'UTR wild type recognition element.Conclusion prestimulation of cerebellar fastigial nucleus has protective effect on cerebral ischemia-reperfusion injury, and its mechanism may be related to the regulation of BIRC2 expression by targeting by miR-29c and the reduction of focal cerebral ischemia-reperfusion injury.
【學(xué)位授予單位】：廣西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2014
【分類號(hào)】：R743.3

【參考文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

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