【摘要】：目的探討抗氧化劑MitoQ對(duì)異氟醚誘導(dǎo)的新生大鼠海馬神經(jīng)元細(xì)胞損傷的影響及潛在機(jī)制。方法 SPF級(jí)健康SD大鼠15只,7日齡,體重15~20g。采用隨機(jī)數(shù)字表法分為三組:對(duì)照組(C組)、異氟醚組(I組)和異氟醚+MitoQ組(IM組),每組5只。C組吸入空-氧混合氣體。Ⅰ組于出生后7、14和21d吸入1.5%異氟醚2h,IM組在每次吸入異氟醚前腹腔注射MitoQ0.4ml/kg。于出生后28d采用HE染色觀察各組大鼠海馬CA1區(qū)海馬神經(jīng)元細(xì)胞形態(tài)。分離培養(yǎng)新生大鼠原代海馬神經(jīng)元細(xì)胞培養(yǎng)并分組處理,采用MTT法和TUNEL原位熒光染色法檢測(cè)細(xì)胞存活率和凋亡率;采用硫代巴比妥酸法和黃嘌呤氧化酶法檢測(cè)細(xì)胞中丙二醛(MDA)濃度和超氧化物歧化酶(SOD)活性;采用Rhodamine 123染色熒光顯微鏡照相法檢測(cè)線粒體膜電位(MMP),DCFH-DA染色熒光顯微鏡照相法檢測(cè)細(xì)胞內(nèi)活性氧簇(ROS)生成量,采用Western blot法檢測(cè)海馬神經(jīng)元細(xì)胞中Bax、Bcl-2和caspase-3蛋白含量。結(jié)果與C組比較,Ⅰ組大鼠海馬組織神經(jīng)細(xì)胞受損明顯,細(xì)胞數(shù)目減少,Ⅰ組細(xì)胞存活率明顯降低,細(xì)胞凋亡率明顯升高,MDA濃度明顯升高,SOD活性明顯降低,ROS生成量明顯增加,MMP水平明顯降低,Bax和caspase-3蛋白含量明顯升高,Bcl-2蛋白含量明顯降低(P0.05);與Ⅰ組比較,IM組大鼠海馬組織神經(jīng)細(xì)胞損傷減少,細(xì)胞存活率明顯升高,細(xì)胞凋亡率明顯降低,MDA濃度明顯降低,SOD活性明顯升高,ROS生成量明顯減少,MMP水平明顯升高,Bax和caspase-3蛋白含量明顯降低,Bcl-2蛋白含量明顯升高(P0.05)。結(jié)論抗氧化劑MitoQ可明顯抑制異氟醚誘導(dǎo)的海馬神經(jīng)元細(xì)胞損傷,這與其拮抗細(xì)胞氧化應(yīng)激和維持線粒體功能作用密切相關(guān)。
[Abstract]:Objective to investigate the effect of antioxidant MitoQ on isoflurane induced hippocampal neuron injury in neonatal rats and its potential mechanism. Methods 15 healthy SD rats of SPF grade, 7 days old, weighing 15 g / 20 g. Three groups were randomly divided into three groups: control group (group C), isofluoroether group (group I) and isofluoroether MitoQ group (IM group), with 5 rats in each group. Group C inhaled 1.5% isofluoroether for 2 hours at 7, 14 and 21 days after birth, and IM group received intraabdominal injection of MitoQ0.4ml/kg. before each inhalation of isofluoroether. The morphology of hippocampal neurons in hippocampal CA1 region of rats in each group was observed by HE staining on the 28th day after birth. The primary hippocampal neurons of neonatal rats were cultured and treated in groups. The survival rate and apoptosis rate were detected by MTT and TUNEL in situ fluorescence staining, and the concentration of malondialdehyde (MDA) and the activity of SOD (SOD) in the cells were detected by thiobarbituric acid method and xanthine oxidase method. Rhodamine 123 staining fluorescence microscope photography was used to detect mitochondrial membrane potential (MMP), DCFH-DA staining. Intracellular active oxygen cluster (ROS) production was detected by fluorescence microscope photography, and Bax,Bcl-2 and caspase-3 protein contents in hippocampal neurons were detected by Western blot assay. Results compared with group C, the number of neurons in hippocampal tissue of group 鈪，

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