本文選題：缺氧缺血 + 自噬　； 參考：《中國(guó)當(dāng)代兒科雜志》2017年08期

【摘要】：目的通過(guò)新生大鼠缺氧缺血腦損傷后神經(jīng)元自噬基因和節(jié)律基因的表達(dá),探討缺氧缺血引起神經(jīng)損傷的新機(jī)制。方法將12只Sprague-Dawley大鼠隨機(jī)分為缺氧缺血組和假手術(shù)組,每組6只。采用結(jié)扎并切斷大鼠右側(cè)頸總動(dòng)脈并給予低氧處理的方法建立體內(nèi)缺氧缺血腦損傷模型。Western blot檢測(cè)兩組大鼠大腦皮層和海馬組織節(jié)律蛋白Clock表達(dá)情況。體外培養(yǎng)大鼠神經(jīng)元細(xì)胞,隨機(jī)分為氧糖剝奪(OGD)組和對(duì)照組,OGD組加入無(wú)糖無(wú)血清DMEM培養(yǎng)基模擬細(xì)胞缺血狀態(tài),并給予低氧處理建立體外缺氧缺血腦損傷模型。采用Western blot檢測(cè)兩組不同時(shí)間點(diǎn)自噬相關(guān)蛋白Beclin1和LC3,以及節(jié)律基因Clock蛋白表達(dá)情況。應(yīng)用si RNA技術(shù)抑制神經(jīng)元Clock蛋白表達(dá)后,檢測(cè)Beclin1和LC3的表達(dá)變化。結(jié)果體外培養(yǎng)神經(jīng)元Beclin1和LC3Ⅱ的表達(dá)呈現(xiàn)節(jié)律性波動(dòng);OGD處理后,體外培養(yǎng)神經(jīng)元Beclin1和LC3Ⅱ的表達(dá)隨著時(shí)間的延長(zhǎng)逐漸升高,不再呈現(xiàn)節(jié)律性波動(dòng);與假手術(shù)組相比,缺氧缺血引起大鼠皮層和海馬組織Clock表達(dá)降低(P0.05);體外培養(yǎng)神經(jīng)元經(jīng)OGD處理后,Clock表達(dá)也較對(duì)照組顯著降低(P0.05);與陰性對(duì)照組相比,抑制神經(jīng)元節(jié)律基因Clock表達(dá)后,自噬相關(guān)蛋白Beclin1和LC3Ⅱ的表達(dá)均顯著下降(P0.05)。結(jié)論缺氧缺血引起神經(jīng)元Beclin1和LC3Ⅱ表達(dá)節(jié)律紊亂,其機(jī)制可能與Clock參與調(diào)控Beclin1和LC3Ⅱ的表達(dá)有關(guān)。
[Abstract]:Objective to investigate the expression of autophagy gene and rhythm gene after hypoxic-ischemic brain injury in neonatal rats and to explore the new mechanism of hypoxia-ischemia induced neuronal injury. Methods Twelve Sprague-Dawley rats were randomly divided into hypoxic-ischemic group and sham-operation group with 6 rats in each group. The rat model of hypoxic-ischemic brain injury was established by ligating and cutting off the right common carotid artery and treated with hypoxia. Western blot was used to detect the expression of circadian rhythm protein clock in cerebral cortex and hippocampus of rats in both groups. Rat neuronal cells were cultured in vitro and randomly divided into two groups: the OGD group and the control group. The model of hypoxic-ischemic brain injury was established by hypoxia-induced hypoxia in the control group (OGD group) and the control group (OGD group) by adding the glucose free serum-free DMEM medium to simulate the ischemic state of the cells. The expression of Beclin1 and LC3 and clock protein were detected by Western blot at different time points. The expression of Beclin1 and LC3 was detected by si RNA technique after inhibiting the expression of clock protein in neurons. Results the expression of Beclin1 and LC3 鈪，

本文編號(hào)：2021206