胰岛素样生长因子-1对Aβ_(25～35)诱导的SH-SY5Y细胞损伤的保护作用及机制  

Protective effect of insulin-like growth factor-1 on SH-SY5Y cell injured by Aβ_(25～35) and the mechanism

作　　者：崔春英 刘鹏飞[2] 侯训尧[1] 申超[1] 洪艳[1] 刘雪平[1] CUI Chun-Ying;LIU Peng-Fei;HOU Xun-Yao(Department of Senile Neurology,Shandong Provincial Hospital Affiliated to Shandong University,Jinan 250021,Shandong,China)

机构地区：[1]山东大学附属省立医院老年神经科,山东济南250021 [2]济宁市第一人民医院

出　　处：《中国老年学杂志》2019年第10期2436-2440,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(81371225);山东省科技发展计划项目(2014GSF118056)

摘　　要：目的观察胰岛素样生长因子(IGF)-1对Aβ_(25～35)诱导的人神经母细胞瘤SH-SY5Y细胞损伤的保护作用,并初步探讨其可能机制。方法将SH-SY5Y细胞随机分为对照(Control)组,Aβ_(25～35)处理(Aβ_(25～35))组,IGF-1治疗(Aβ_(25～35)+IGF-1)组,Wortmannin抑制剂(Aβ_(25～35)+IGF-1+Wort)组,单纯渥曼青霉素(Wortmannin)组。CCK-8法检测细胞活力,检测乳酸脱氧酶(LDH)漏出率,判断细胞损伤程度,荧光探针DCFH-DA法检测细胞内活性氧(ROS)含量的变化,试剂盒测定丙二醛(MAD)含量及超氧化物气化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)活性,流式细胞仪分析法测定细胞凋亡率,Western印迹法检测相关蛋白表达。结果与Aβ_(25～35)组相比,Aβ_(25～35)+IGF-1组细胞活力明显升高,LDH漏出率明显下降(P<0.05)。IGF-1可明显减少Aβ_(25～35)诱导SH-SY5Y细胞ROS及MDA生成,增加SOD及GSH-Px酶活性(P<0.05)。与Aβ_(25～35)组相比,IGF-1治疗还能增加蛋白激酶B(Akt)及叉头框蛋白(Fox)O3a磷酸化水平,降低Puma蛋白表达水平及细胞凋亡率(P<0.05)。而加入磷脂酰肌醇-3激酶(PI3K)特异性抑制剂Wortmannin预处理,可阻断上述IGF-1的保护作用(P<0.05)。结论 IGF-1对Aβ_(25～35)诱导的SH-SY5Y细胞损伤的保护作用可能与激活PI3K/Akt信号通路有关。Objective To investigate the protective effect of insulin-like growth factor (IGF)-1 on human neuroblastoma SH-SY5Y cells injured by Aβ25~35 and the possible mechanism. Methods SH-SY5Y cells were randomly divided into control, Aβ25~35 , IGF-1 treatment (Aβ25~35+IGF-1), Wortmannin inhibitor (Aβ25~35+IGF-1+Wort), simple Wortmannin(Wortmannin) groups. The cell viability was detected by CCK-8 assay. The degree of the cell damage was evaluated by determining lactate dehydrogenase (LDH) leakage. The reactive oxygen species (ROS) level was evaluated by DCFH-DA fluorescent probe. The related kits were used for detecting MDA content, SOD and GSH-Px activities. The apoptosis rate of cell was measured by flow cytometry. The expressions of proteins were detected by Western blot. Results Compared with those of Aβ25~35 group, the cell viability was significantly increased, and the leakage rate of LDH significantly was decreased in Aβ25~35+IGF-1 group ( P <0.05). IGF-1 significantly decreased the productions of ROS and MDA,and increased the activities of SOD and GSH-Px in SH-SY5Y cells ( P <0.05). Compared with that of Aβ25~35 group, IGF-1 also significantly increased the phosphorylation levels of Akt and FoxO3a, and decreased the Puma protein expression and cells apoptosis rate (P <0. 05). But all of above protective effect of IGF-1 was blocked by PI3K specific inhibitor Wortmannin. Conclusions The protective effect of IGF -1 on SH-SY5Y cell injured by Aβ25~35 might be related to the activation of PI3K/Akt signaling pathway.

关 键 词：胰岛素样生长因子-1 Β淀粉样蛋白 蛋白激酶B 氧化应激 凋亡 

分 类 号：R741.05[医药卫生—神经病学与精神病学]