N-乙酰-L-色氨酸减轻H_2O_2诱导小鼠海马神经元细胞凋亡  

N-acetyl-L-tryptophan alleviates the apoptosis induced by H_2O_2 of mice PHNs

作　　者：于树娜[1] 王志芳[2] 杜晓东[1] 姜红心[3] 李进[1] 史才兴[1] 蒋吉英[1] 

机构地区：[1]潍坊医学院人体解剖学教研室,山东潍坊261053 [2]即墨市人民医院神经内科,山东郎墨266200 [3]潍坊医学院形态学实验室,山东潍坊261053

出　　处：《基础医学与临床》2013年第4期423-428,共6页Basic and Clinical Medicine

基　　金：山东省"泰山学者"建设工程专项经费自主项目;山东省自然科学基金(ZR2010HM006);山东省教育厅资助项目(J07WE27)

摘　　要：目的探讨N-乙酰-L-色氨酸(L-NAT)对海马神经元(PHN)缺血低氧损伤的影响。方法用600μmol/L H2O2诱导PHN制备海马神经元细胞凋亡模型,采用免疫荧光染色检测caspase-3的表达,Rhodamine 123染色检测线粒体膜势能(ΔΨm)的改变,锥虫蓝染色检测细胞存活率,比色法检测caspase-3、乳酸脱氢酶(LDH)的活性,Westernblot检测caspase-3及凋亡诱导因子(AIF)和细胞色素C(CytC)等线粒体促凋亡因子在胞质蛋白和线粒体蛋白中的表达。结果 L-NAT可减轻H2O2所引起的细胞形态的死亡、存活率的降低、LDH的释放、caspase-3的激活、线粒体膜势能的丧失及AIF和CytC等线粒体促凋亡因子的释放。结论 L-NAT能通过抑制caspase依赖性和非依赖性的细胞凋亡途径,减轻H2O2诱导的小鼠海马神经元的细胞损伤。Objective To investigate the effects of N-acetyl-L-tryptophan （L-NAT） on H2O2-induced apoptosis of primary hippoeampal neurons （PHN）. Methods An apoptosis cellular model of cerebral ischemia was made by H2O2. Immunofluorescence staining was used to detect the expression of caspase-3, Rhodamine 123 staining was used to detect mitochondrial membrane potential changes, trypan blue staining was used to detect survival rate, the colorimetric assay was used to detect the activity of caspase-3 and LDH, Western blot was used to detect the expres- sions of caspase-3 and apoptosis-inducing factor and cytochrome C and other mitochondrial pro-apoptotie factor in the cytosolic proteins and mitochondrial proteins. Results L-NAT inhibited H2O2-induced cell death, loss of mito- chondrial membrane potential, downstream release of LDH and mitochondrial factors, and activation of caspase-3 in primary hippoeampal neurons. Conclusions L-NAT has the effectiveness on cerebral ischemia/hypoxia injury though caspase-independent caspase-dependent apoptosis pathway.

关 键 词：N-乙酰-L-色氨酸 神经元 缺血低氧损伤 细胞凋亡 细胞模型 

分 类 号：R363[医药卫生—病理学]