■在这项研究中,研究了2-花生酰甘油2-AG对MCAO后小鼠海马神经元非caspase依赖性凋亡的神经保护机制。
■将150只健康清洁雄性C57BL/6小鼠随机分为3组:假手术组,模型组和2-AG治疗组,每组50只小鼠。采用改良的ZeaLonga法建立小鼠大脑中动脉闭塞(MCAO)模型。流式细胞术检测海马神经细胞凋亡率和线粒体膜电位。AIF的mRNA表达,通过qPCR测定海马组织中的EndoG和BNIP3。Westernblot检测AIF蛋白表达,海马组织线粒体中的EndoG和BNIP3。
■2-AG治疗组海马神经元凋亡率明显低于模型组(P<0.01),表明2-AG可抑制MCAO诱导的海马神经元凋亡。然而,2-AG处理组海马神经元线粒体膜电位明显高于模型组(P<0.01),提示2-AG能提高MCAO小鼠海马神经元线粒体膜电位。实时定量PCR(qPCR)显示2-AG能抑制AIF基因的表达,海马组织中的EndoG和BNIP3。Westernblot结果显示,2-AG能抑制AIF的分泌,内G和BNIP3进入线粒体的细胞质。
■内源性大麻素2-AG对神经元损伤有保护作用,其机制可能与海马区的脑神经细胞的保护和线粒体功能的完整性有关。内源性大麻素2-AG可能抑制非caspase依赖性细胞凋亡途径,从而发挥其神经保护作用。
UNASSIGNED: In this study, the neuroprotective mechanism of 2-arachidonoylglycerol 2-AG against non-caspase-dependent apoptosis in mice hippocampal neurons following MCAO was investigated.
UNASSIGNED: One hundred and fifty healthy clean male C57BL/6 mice were randomly divided into 3 groups: sham group, model group and 2-AG treatment group, 50 mice in each group. A modified Zea Longa method was used to establish a model of middle cerebral artery occlusion (MCAO) in mice. The apoptosis rate and mitochondrial membrane potential of hippocampal nerve cells were measured by flow cytometry. The mRNA expressions of AIF, Endo G and BNIP3 in hippocampal tissues were determined by qPCR. Western blot was used to determine the protein expressions of AIF, Endo G and BNIP3 in the mitochondria of hippocampal tissue.
UNASSIGNED: The apoptosis rate of hippocampal neurons in the group treated with 2-AG was significantly lower than that of the model (P<0.01), which indicated that 2-AG could inhibit the apoptosis of hippocampal neurons induced by MCAO. However, the mitochondrial membrane potential of hippocampal neurons in the group treated with 2-AG was significantly higher than that of the model (P<0.01), indicating that 2-AG could improve the mitochondrial membrane potential of hippocampal neurons in MCAO mice. Real-time quantitative PCR (qPCR) showed that 2-AG could inhibit the gene expressions of AIF, Endo G and BNIP3 in hippocampal tissues. Western blot results showed that 2-AG could inhibit the secretions of AIF, Endo G and BNIP3 into cytoplasm in mitochondria.
UNASSIGNED: Endocannabinoids 2-AG had a protective effect on neurons injury, and the mechanism was possibly associated with the protection of the brain nerve cells in the hippocampus and the integrity of the mitochondrial function. Endocannabinoids 2-AG may inhibit the non-caspase-dependent apoptosis pathway, so as to exert its nerve protective effect.