外源性硫化氢对原代神经元早老素1 和β 淀粉样蛋白的影响.docVIP

外源性硫化氢对原代神经元早老素1和β淀粉样蛋白的影响 陈筱山，何选丽，朱丽娟，张华，冯飞，晏勇 (400016重庆，重庆医科大学附属第一医院神经内科) [摘要] 目的 探讨外源性硫化氢（hydrogen Sulfide,H2S）对原代培养皮质神经元早老素1 (presensilin1，PS1)和β淀粉样蛋白(β-amyloid peptide, Aβ)的影响及机制。方法 ①以硫氢化钠（sodium hydrosulfide，NaHS）为外源性H2S的供体，不同浓度NaHS（0、5、10、20、30、40、50μmol/L）作用于原代培养的皮质神经元，TUNEL法检测细胞凋亡，酶联免疫吸附试验(ELISA)检测Aβ1-42水平，Western blot检测PS1蛋白表达水平；②神经元分为对照组（0μmol/L NaHS）、20μmol/L NaHS组、A干预组（NaHS 20μmol/L加30min PI3K通路阻断剂LY294002 20μmol/L）和B干预组（NaHS 20μmol/L加30min MAPK通路阻断剂PD98059 20μmol/L），Western blot检测PS1蛋白表达。结果 ①5~20μmol/L浓度NaHS干预时，不引起明显神经元凋亡；30~50μmol/L时，神经元凋亡明显增加（P0.05）；10、20μmol/L浓度NaHS显著降低 Aβ1-42水平（P0.05）；10~20μmol/L浓度NaHS显著降低 PS1蛋白表达水平（P0.05）。②与对照组相比，在20μmol/L NaHS组和B干预组，PS1蛋白表达降低（P0.05），而在A干预组，PS1蛋白表达无改变（P0.05）；而20μmol/L NaHS组和B干预组间无差异（P0.05）；与A干预组相比，B干预组PS1蛋白表达显著降低（P0.05）。结论 低浓度外源性H2S(30μmol/L浓度NaHS范围内)不引起神经元明显凋亡，降低Aβ1-42水平和PS1蛋白表达，并可能通过PI3K信号通路发挥降低PS1蛋白表达的作用。 [关键词] 硫化氢；原代神经元；早老素1 Effects of Exogenous Hydrogen Sulfide on the expression of Presenilin1 and Aβ in Primary Culture Neuron Chen Xiaoshan，He Xuanli，Zhu Lijuan，Zhang Hua，Feng Fei，Yan Yong （Department of Neurology, the first Affiliated Hospital of Chongqing Medical University，Chongqing，China，400016） [Abstract] Objective To investigate the exogenous Hydrogen Sulfide（H2S）on the expression of Presenlin1(PS1)and Aβ in primary culture neuron. Methods NaHS was used as H2S donor，different concentration(0、5、10、20、30、40、50μmol/L )of NaHS was added to primary cortical neurons, apoptosis rate was measured by TUNEL，Aβ1-42 level was measured by ELISA，the expression of PS1 protein level was measured by Western blot. The neurons was then devided into four groups:control(0μmol/L NaHS),20 μmol/L NaHS, A group(the PI3K signal inhibitor LY294002 20μmol/L+ 20μmol/L NaHS ),B group(the MAPK signal inhibitor PD 98059 20μmol/L+ 20μmol/L NaHS ) Results The apoptosis rate was significantly increased（P0.05）in the concentration of 30~50umol/L NaHS; Aβ1-42 concentration was decreased（P0.05）in concentration of 10~20μmol/L of NaHS.PS1 pr