【摘要】：目的探討褪黑素通過(guò)沉默信息調(diào)節(jié)因子1(SIRT1)減輕線粒體氧化應(yīng)激機(jī)制對(duì)腦缺血-再灌注(IR)小鼠的作用。方法通過(guò)線栓法建立小鼠短暫性大腦中動(dòng)脈阻塞(MCAO)IR模型,對(duì)190只小鼠經(jīng)腹腔注射褪黑素和側(cè)腦室注射SIRT1抑制劑EX527,剔除死亡小鼠30只,按隨機(jī)數(shù)字表法分為IR組、褪黑素組、褪黑素+EX527組、EX527組各40只,通過(guò)2,3,5-氯化三苯基四氮唑(TTC)法測(cè)定腦梗死體積、干濕比重法測(cè)定腦水腫并進(jìn)行神經(jīng)功能缺損評(píng)分。通過(guò)Western blot檢測(cè)線粒體和胞質(zhì)SIRT1、Ac-P53、Ac-核因子κB(Ac-NF-κB)、BCL2、BAX蛋白以及細(xì)胞色素C蛋白表達(dá)。多組間比較采用單因素方差分析。結(jié)果 (1)4組間腦梗死體積、神經(jīng)功能障礙評(píng)分和腦水腫的差異均有統(tǒng)計(jì)學(xué)意義(F值分別為16.452、23.622和18.786,均P0.05)。4組SITR1、Ac-P53、Ac-NF-κB、BCL2、BAX表達(dá)差異均有統(tǒng)計(jì)學(xué)意義(F值分別為2 348.158、1 434.841、7 042.563、14 627.128和691.475,均P0.05)。4組小鼠線粒體膜電位、線粒體活性氧和復(fù)合體Ⅰ活性差異有統(tǒng)計(jì)學(xué)意義(F值分別為28.454、33.728和29.716,均P0.05)。(2)與IR組相比,褪黑素組的腦梗死體積縮小(32±5)mm~3比(57±5)mm~3,P0.05);神經(jīng)功能障礙評(píng)分降低(2.4±0.3比3.5±0.3,P0.05);腦水腫減輕(80.2±0.9)%比(83.9±1.2)%,P0.05);SIRT1和抗凋亡蛋白BCL2的表達(dá)增加(P0.05);促凋亡蛋白BAX和Ac-P53、Ac-NF-κB的表達(dá)減少(P0.05);線粒體膜電位、線粒體復(fù)合體Ⅰ活性和細(xì)胞色素C水平提高(P0.05);胞質(zhì)活性氧產(chǎn)物和細(xì)胞色素C水平降低(P0.05)。(3)與褪黑素組相比,褪黑素+EX527組的腦梗死體積增大[(42±5)mm~3比(32±5)mm~3,P0.05];神經(jīng)功能障礙評(píng)分增高(3.2±0.3比2.4±0.3,P0.05);腦水腫加重[(83.4±0.8)%比(80.2±0.9)%,P0.05];SIRT1和抗凋亡蛋白BCL2的表達(dá)減少(P0.05);促凋亡蛋白BAX和Ac-P53、Ac-NF-κB的表達(dá)增加(P0.05);線粒體膜電位、線粒體復(fù)合體Ⅰ活性和細(xì)胞色素C水平降低(P0.05);胞質(zhì)活性氧產(chǎn)物和胞質(zhì)細(xì)胞色素C水平提高(P0.05)。結(jié)論在缺血性卒中模型小鼠中,褪黑素通過(guò)激活SIRT1信號(hào)通路,減輕線粒體氧化應(yīng)激損傷和細(xì)胞死亡,從而發(fā)揮腦保護(hù)作用。
[Abstract]:Objective to investigate the protective effect of melatonin on cerebral ischemia-reperfusion (IR) mice by silencing information regulatory factor 1 (SIRT1). Methods the (MCAO) IR model of transient middle cerebral artery occlusion in mice was established by the method of thread occlusion. Thirty dead mice were killed by intraperitoneal injection of melatonin and injection of SIRT1 inhibitor EX527,. According to the method of random number table, they were divided into IR group and melatonin group. The volume of cerebral infarction was measured by (TTC) method in melatonin EX527 group and 40 rats in EX527 group. Brain edema was measured by dry-wet specific gravity method and neurological impairment score was evaluated. The expression of SIRT1,Ac-P53,Ac- nuclear factor 魏 B (Ac-NF- 魏 B), BCL2,BAX protein and cytochrome C protein) in mitochondria and cytoplasm was detected by Western blot. Single factor analysis of variance (ANOVA) was used for multi-group comparison. Results (1) the volume of diencephalic infarction in 4 groups, There were significant differences in neurological dysfunction score and cerebral edema (F = 16.452, 23.622 and 18.786, respectively, P0.05). There were significant differences in the expression of BCL2BAX in SITR1,Ac-P53,Ac-NF- 魏 B, BCL2BAX in 4 groups (F = 2 348.158 1 434.841 7 042.56314 627.128 and 691.475, respectively, P0.05). The activity of reactive oxygen species (Ros) and complex 鈪，

本文編號(hào)：2285104