垂体腺苷酸环化酶激活肽拮抗lactacystin细胞毒性作用的机制研究  

Effect of pituitary adenylate cyclase activating peptide on cytotoxicity of lactacystin and its mechanism

作　　者：管丽娜[1] 巴茂文[2] 冀永强[3] 连培文[4] 李宁[4] 牟贤玉 于国平[2] 

机构地区：[1]烟台毓璜顶医院急诊科,264000 [2]烟台毓璜顶医院神经内科,264000 [3]烟台毓璜顶医院肾病内科,264000 [4]烟台毓璜顶医院中心实验室,264000

出　　处：《中华老年心脑血管病杂志》2015年第4期421-424,共4页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases

基　　金：山东省自然科学基金(ZR2010HM117);烟台市科技计划项目(2010148-6)

摘　　要：目的探讨垂体腺苷酸环化酶活化肽(pituitary adenylate cyclase-activating polypeptide,PACAP)对蛋白酶体抑制剂lactacystin诱导的帕金森病多巴胺能细胞凋亡作用的影响及其分子机制。方法用神经生长因子将鼠嗜铬细胞瘤PC12细胞诱导分化成神经元的细胞模型,经20μmol/L的lactacystin作用24h,建立帕金森病细胞实验模型。实验分为对照组、lactacystin组、PACAP1-27干预组(干预1组)、PACAP1-27和PACAP6-27共同干预组(干预2组)。Western blot法检测线粒体凋亡相关蛋白bcl-2、bax、bcl-2/bax比值、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)的表达情况。结果与对照组比较,lactacystin组bcl-2表达、bcl-2/bax比值明显降低(P<0.01),bax表达无明显变化,Caspase-3表达明显升高(P<0.01);与lactacystin组比较,干预1组bcl-2表达、bcl-2/bax比值明显升高(P<0.01),bax表达无变化,Caspase-3表达明显降低(P<0.01);与干预1组比较,干预2组Caspase-3表达明显升高(P<0.01)。结论蛋白酶体抑制剂lactacystin引起线粒体损伤导致细胞损伤;PACAP1-27通过调节上述信号通路发挥保护作用。而PACAP1-27的受体拮抗剂PACAP6-27则减弱了PACAP1-27的这一作用。Objective To study the effect of pituitary adenylate cyclase activating peptide（PACAP）on lactacystin-induced dopaminergic apoptosis in Parkinson disease and its molecular mechanism.Methods A model of Parkinson＇s disease cells was established by differentiating pheochromocyte PC12 cells into neurons with nerve growth factor treated with lactacystin（20μmol/L）for 24 h.The cells were divided into control group,lactacystin treatment group,PACAP1-27 treatment group,PACAP1-27＋PACAP6-27 treatment group.Expressions of bcl-2,bax,Caspase-3and ratio of bcl-2/bax were detected by Western blot.Results The expression level of bcl-2and the ratio of bcl-2/bax were significantly lower in lactacystin treatment group than in control group and significantly higher in PACAP1-27 treatment group and PACAP1-27＋PACAP6-27 treatment group than in lactacystin treatment group（P〈0.01）with no significant difference observed in the expression level of bax.The expression level of Caspase-3 was significantly higher in lactacystin treatment group than in control group and in PACAP1-27＋PACAP6-27 treatment group than in PACAP1-27 treatment group,and was significantly lower in PACAP1-27 treatment group than in lactacystin treatment group（P〈0.01）.Conclusion Lactacystin-induced mitochondrial damage can lead to cell damage.PACAP1-27 protects cells against damage by regulating their signal pathway,which is attenuated by PACAP6-27 which is a PACAP1-27 receptor antagonist.

关 键 词：受体 垂体腺苷酸环化酶激活多肽 帕金森病 多巴胺 细胞凋亡 PC12细胞 半胱氨酸天冬氨酸蛋白酶3 

分 类 号：R742.5[医药卫生—神经病学与精神病学]