抗氧化剂MitoQ对异氟醚诱导大鼠海马神经元损伤的影响及机制
发布时间：2019-06-22 19:50
【摘要】：目的探讨抗氧化剂MitoQ对异氟醚诱导的新生大鼠海马神经元细胞损伤的影响及潜在机制。方法 SPF级健康SD大鼠15只,7日龄,体重15~20g。采用随机数字表法分为三组:对照组(C组)、异氟醚组(I组)和异氟醚+MitoQ组(IM组),每组5只。C组吸入空-氧混合气体。Ⅰ组于出生后7、14和21d吸入1.5%异氟醚2h,IM组在每次吸入异氟醚前腹腔注射MitoQ0.4ml/kg。于出生后28d采用HE染色观察各组大鼠海马CA1区海马神经元细胞形态。分离培养新生大鼠原代海马神经元细胞培养并分组处理,采用MTT法和TUNEL原位荧光染色法检测细胞存活率和凋亡率;采用硫代巴比妥酸法和黄嘌呤氧化酶法检测细胞中丙二醛(MDA)浓度和超氧化物歧化酶(SOD)活性;采用Rhodamine 123染色荧光显微镜照相法检测线粒体膜电位(MMP),DCFH-DA染色荧光显微镜照相法检测细胞内活性氧簇(ROS)生成量,采用Western blot法检测海马神经元细胞中Bax、Bcl-2和caspase-3蛋白含量。结果与C组比较,Ⅰ组大鼠海马组织神经细胞受损明显,细胞数目减少,Ⅰ组细胞存活率明显降低,细胞凋亡率明显升高,MDA浓度明显升高,SOD活性明显降低,ROS生成量明显增加,MMP水平明显降低,Bax和caspase-3蛋白含量明显升高,Bcl-2蛋白含量明显降低(P0.05);与Ⅰ组比较,IM组大鼠海马组织神经细胞损伤减少,细胞存活率明显升高,细胞凋亡率明显降低,MDA浓度明显降低,SOD活性明显升高,ROS生成量明显减少,MMP水平明显升高,Bax和caspase-3蛋白含量明显降低,Bcl-2蛋白含量明显升高(P0.05)。结论抗氧化剂MitoQ可明显抑制异氟醚诱导的海马神经元细胞损伤,这与其拮抗细胞氧化应激和维持线粒体功能作用密切相关。
[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，
本文编号：2504905
[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，
本文编号：2504905
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