氯化锂对冈田酸所致SK-N-SH神经元突触萎缩的保护作用  

Protective effects of LiCl on synaptic atrophy induced by okadaic acid in differentiated SK-N-SH neuroblastoma cells

作　　者：袁玉婷[1] 佀营营 王占洋[1] 弥相权[3] 吕欢欢[4] 许波[2] 李刚[4] 李忌[2] 何杰[2] 王振华[2] 

机构地区：[1]石河子大学药学院,新疆石河子832005 [2]烟台大学生命科学学院线粒体与健康衰老研究中心,山东烟台264005 [3]兰州大学生命科学学院,甘肃兰州730000 [4]中国科学院西北高原生物研究所,青海西宁810001

出　　处：《中国药理学通报》2016年第5期670-675,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金面上项目(No 11175222;21372190);石河子大学重大科技攻关计划项目(No gxjs2012-zdgg02);山东省泰山学者建设工程专项经费(No tshw201502046);青海省科技创新能力促进计划项目(No 2014-ZJ-765);烟台市科技计划项目(No 2014LGS004)

摘　　要：目的考察氯化锂(LiCl)对蛋白磷酸酶抑制剂冈田酸(OA)诱导的SK-N-SH细胞分化神经元损伤的保护作用和tau蛋白Ser-^(262)位点磷酸化水平的影响。方法利用全反式维甲酸(ATRA)诱导SK-N-SH细胞分化为成熟的神经元细胞;采用OA诱导成熟神经元细胞建立AD模型;采用磺酰罗丹明B(SRB)比色法考察LiCl对成熟的神经元细胞增殖的抑制作用;Giemsa染色观察SK-N-SH细胞形态学变化;并采用Image-Proplus软件测定神经元细胞的突触长度;采用Western blot检测synaptophysin蛋白和tau蛋白Ser-^(262)位点磷酸化水平。结果 10μmol·L^(-1)ATRA连续处理7 d,可诱导SK-N-SH细胞突触生长和synaptophysin蛋白表达等典型分化神经元的特征。20~100 nmol·L^(-1)OA作用于分化神经元,可浓度和时间依赖性抑制细胞增殖,同时致分化神经元突触萎缩,tau蛋白Ser-^(262)位点磷酸化水平也明显升高。10 mmol·L^(-1)LiCl预处理可维持synaptophysin蛋白高表达,抑制tau蛋白Ser-^(262)位点磷酸化水平(P<0.01)。结论LiCl能够改善OA所致分化神经元的突触损伤,并伴随着synaptophysin表达的升高、tau蛋白Ser-^(262)位点异常磷酸化水平的降低。Aim To explore the protective effects of lithium chloride ( LiCl ) on neurous injuries and phos-phorylation of tau protein at serine262 induced by okada-ic acid( OA) . Methods The neuroblastoma SK-N-SH cells were differentiated by all-trans-retinoic acid ( AT-RA) . The differentiated SK-N-SH cells were treated with OA to establish the Alzheimer′s disease cellular model. SK-N-SH cells′ viability and proliferation were measured by SRB test. Giemsa staining was used to observe cell morphology. The neurite length of SK-N-SH cells was measured by Image-Proplus software. Syn-aptophysin and phosphorylated tau protein at serine262 expression levels were tested by Western blot. Results The SK-N-SH cells which were treated with 10 μmol ·L-1 ATRA for 7 days displayed mature neuronal fea-tures. The synaptic length of SK-N-SH cells became longer. And the levels of serine262 phospho-tau was sig-nificantly elevated. 20~100 nmol·L-1 OA effectively&nbsp;inhibited the viability of differentiated SK-N-SH cells in a concentration-dependent manner and in a time-de-pendent manner. The OA treatment induced obvious synaptic atrophy in differentiated SK-N-SH cells. And the phosphorylation level of tau protein serine262 also greatly increased. The pretreatment with 10 mmol · L-1 LiCl significantly ameliorated the synaptic atrophy, the decrease of synaptophysin expression and the in-crease of tau phosphorylation at serine262 induced by OA in differentiated SK-N-SH cells. Conclusion LiCl could effectively inhibit OA-induced synaptic atro-phy in differentiated SK-N-SH cells, and it could also result in the increase of synaptophysin expression and the decrease of the phosphorylation of tau protein at serine262 .

关 键 词：LICL 冈田酸 TAU蛋白磷酸化 阿尔茨海默病 分化神经元 突触萎缩 吉姆萨染色 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R338.13[医药卫生—基础医学]