本文選題：缺氧缺血 + 自噬��； 參考：《中國當代兒科雜志》2017年08期

【摘要】：目的通過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,探討缺氧缺血引起神經(jīng)損傷的新機制。方法將12只Sprague-Dawley大鼠隨機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檢測兩組大鼠大腦皮層和海馬組織節(jié)律蛋白Clock表達情況。體外培養(yǎng)大鼠神經(jīng)元細胞,隨機分為氧糖剝奪(OGD)組和對照組,OGD組加入無糖無血清DMEM培養(yǎng)基模擬細胞缺血狀態(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檢測兩組不同時間點自噬相關(guān)蛋白Beclin1和LC3,以及節(jié)律基因Clock蛋白表達情況。應用si RNA技術(shù)抑制神經(jīng)元Clock蛋白表達后,檢測Beclin1和LC3的表達變化。結(jié)果體外培養(yǎng)神經(jīng)元Beclin1和LC3Ⅱ的表達呈現(xiàn)節(jié)律性波動;OGD處理后,體外培養(yǎng)神經(jīng)元Beclin1和LC3Ⅱ的表達隨著時間的延長逐漸升高,不再呈現(xiàn)節(jié)律性波動;與假手術(shù)組相比,缺氧缺血引起大鼠皮層和海馬組織Clock表達降低(P0.05);體外培養(yǎng)神經(jīng)元經(jīng)OGD處理后,Clock表達也較對照組顯著降低(P0.05);與陰性對照組相比,抑制神經(jīng)元節(jié)律基因Clock表達后,自噬相關(guān)蛋白Beclin1和LC3Ⅱ的表達均顯著下降(P0.05)。結(jié)論缺氧缺血引起神經(jīng)元Beclin1和LC3Ⅱ表達節(jié)律紊亂,其機制可能與Clock參與調(diào)控Beclin1和LC3Ⅱ的表達有關(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：2021206