番茄红素对原代小鼠皮质神经元的保护作用及机制研究

Lycopene protects primary mouse cerebrocortical neurons against t-BHP-induced damage in vitro

目的:观察番茄红素(lycopene)对氧化损伤的原代皮质神经元的保护效应并探讨其作用机制。方法:采用原代细胞培养技术体外分离培养小鼠皮质神经元,通过免疫荧光染色法检测微管相关蛋白2(microtubuleassociated protein 2,MAP-2)的表达进行鉴定。将神经元分为4组:正常神经元组、叔丁基过氧化氢(tert-butyl hydroperoxide,t-BHP)处理组、t-BHP+lycopene处理组和lycopene处理组,培养24 h,采用MTT法检测各组神经元的活力;采用流式细胞技术检测各组神经元内ROS的水平;Western blot法检测各组神经元Bax、Bcl-2、caspase-3、cleaved caspase-3及细胞色素C蛋白表达的变化。结果:Lycopene能明显提高t-BHP处理的神经元活性,降低tBHP处理的神经元内ROS含量,同时上调Bcl-2蛋白的表达,降低Bax、cleaved caspase-3和细胞色素C蛋白的表达(P<0.05)。结论:Lycopene能够对抗t-BHP诱导的原代皮质神经元的损伤,抑制神经元凋亡,其机制可能与降低神经元内ROS的水平及上调的表达有关。

AIM： To investigate the protective effect of lycopene on primary mouse cerebrocortical neurons ex-posed to tert-butyl hydroperoxide （t-BHP） and its mechanisms of in vitro. METHODS： Primary cerebrocortical neurons of newborn C57 mice were extracted and divided into normal group, t-BHP group, lycopene ＋ t-BHP group and lycopene group. The neuronal damage was induced by t-BHP exposure for 24 h, and the cell viability was examined by MTT assay.ROS content was measured by flow cytometry, and the protein levels of Bax, Bcl-2 , caspase-3 , cleaved caspase-3 and cyto-chrome C were examined by Western blot. RESULTS： The primary mouse cortical neurons expressed MAP-2 protein. Ly-copene at concentration of 4 jjimol/L reversed the decrease in cell viability. Flow cytometry revealed that lycopene treatment attenuated ROS content under the condition of t-BHP exposure. In addition,the protein level of Bcl-2 was increased, and the expression of Bax, cleaved caspase-3 and cytochrome-C was suppressed in lycopene ＋ t-BHP group. CONCLUSION：The protective effect of lycopene on cortical neurons with t-BHP-induced injury may be involved in the mechanism of neuro-nal antioxidative response by down-regulating caspase-3 and Bax/Bcl-2 through the mitochondrial apoptotic pathway.