發(fā)布時(shí)間：2018-10-12 08:30

【摘要】：目的觀察睡眠剝奪(sleep deprivation,SD)對(duì)內(nèi)側(cè)前額葉皮層(medial prefrontal cortex,m PFC)神經(jīng)元自噬(autophagy)活動(dòng)的影響及機(jī)制。方法實(shí)驗(yàn)使用成年(8~10周)雄性C57BL/6小鼠,體重23-26g,隨機(jī)分為睡眠剝奪6 h(SD6 h)組與其對(duì)照組(n=10-12只/組)、睡眠剝奪12 h(SD12 h)組與其對(duì)照組(n=10-12只/組)。采用輕柔刺激法建立SD6 h和SD12 h模型。蛋白免疫印跡(Western blot)檢測(cè)內(nèi)側(cè)前額葉神經(jīng)元皮層超氧化物歧化酶2(superoxide dismutase,SOD2)、過(guò)氧化氫酶(catalase)、沉默信息調(diào)節(jié)因子3(Silent information regulator 3,Sirt3)、微管相關(guān)蛋白1輕鏈3-Ⅱ(LC3-Ⅱ)、P62(也稱SQSTM1)、Beclin-1、腺苷酸活化蛋白激酶(AMP-activated protein kinase,AMPK)、雷帕霉素靶蛋白(mammalian target of rapamycin,m TOR)、p-AMPK、p-m TOR蛋白的表達(dá)水平。免疫熒光染色觀察內(nèi)側(cè)前額葉皮層神經(jīng)元溶酶體膜相關(guān)蛋白1(lysosomal-associated membrane protein 1,LAMP-1)的表達(dá)情況,Dihydroethidine(DHE)染色檢測(cè)m PFC神經(jīng)元O2-的水平。結(jié)果1.在正常睡眠覺(jué)醒周期中,與白天相比,夜間自噬流活性較高,P62蛋白表達(dá)較低,Beclin-1蛋白表達(dá)相對(duì)較高。2.與對(duì)照組相比,SD6 h后m PFC神經(jīng)元SOD2(P0.05)、Catalase(P0.05)和Sirt3(P0.05)蛋白表達(dá)均顯著上調(diào),O2-的水平同時(shí)也相對(duì)升高(P0.05);而SD12 h后,m PFC神經(jīng)元SOD2、Catalase和Sirt3蛋白表達(dá)水平與對(duì)照組相比均無(wú)顯著差異,但O2-的水平相對(duì)升高。3.與對(duì)照組相比,SD6 h與SD12 h均使m PFC神經(jīng)元LC3-Ⅱ(P0.05)和P62的表達(dá)水平升高(P0.05),進(jìn)一步研究發(fā)現(xiàn),Beclin-1(P0.05)和LAMP-1(P0.05)的表達(dá)水平均相對(duì)升高。4.SD6 h后,與對(duì)照組相比,m PFC神經(jīng)元AMPK和m TOR蛋白水平無(wú)顯著變化;SD12 h后,與對(duì)照組相比,m PFC神經(jīng)元m TOR蛋白表達(dá)水平顯著下調(diào)(P0.05),但AMPK蛋白表達(dá)無(wú)明顯變化。結(jié)論1.睡眠期間,m PFC神經(jīng)元自噬活動(dòng)較低,而覺(jué)醒期間m PFC神經(jīng)元自噬活動(dòng)較高。2.睡眠剝奪后,m PFC神經(jīng)元氧化水平升高,而抗氧化水平只產(chǎn)生一過(guò)性升高。3.無(wú)論是SD6 h還是SD12 h,均使m PFC神經(jīng)元自噬活動(dòng)增強(qiáng)。4.SD12 h使m PFC神經(jīng)元m TOR信號(hào)通路被抑制,進(jìn)而誘導(dǎo)自噬活動(dòng)上調(diào)。
[Abstract]:Objective to investigate the effect and mechanism of sleep deprivation (sleep deprivation,SD) on autophagy (autophagy) activity of (medial prefrontal cortex,m PFC neurons in medial prefrontal cortex. Methods male C57BL/6 mice were randomly divided into sleep deprivation group (SD6 h) and control group (n = 10-12), sleep deprivation for 12 h (SD12 h) and control group (n = 10-12). SD6 h and SD12 h models were established by soft stimulation. Western blot (Western blot) detection of superoxide dismutase 2 (superoxide dismutase,SOD2), catalase (catalase), silencing signaling factor 3 (Silent information regulator 3 sirt3, microtubule-associated protein 1 light chain 3- 鈪，

本文編號(hào)：2265469