阿利吉仑对氧糖剥夺损伤的SH-SY5Y细胞的保护作用及可能机制  

Protective effect of aliskiren on SH-SY5Y cells injured by oxygen-glucose deprivation and its possible mechanisms

作　　者：陆婉杏[1,2] 蒙兰青[1] 黄晓华[1] LU Wan-xing;MENG Lan-qing;HUANG Xiao-hua(Department of Neurology, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, China;Department of Neurology, Minzu Hospital of Guangxi Zhuang Autonomous Region, Nanning 530001, China)

机构地区：[1]右江民族医学院附属医院神经内科,广西百色533000 [2]广西壮族自治区民族医院神经内科,广西南宁530001

出　　处：《中国病理生理杂志》2019年第9期1630-1634,共5页Chinese Journal of Pathophysiology

基　　金：广西高校科学技术研究项目(No.YB2014301)

摘　　要：目的:探讨阿利吉仑对氧糖剥夺(OGD)损伤的神经母细胞瘤SH-SY5Y细胞的保护作用及可能机制。方法:将SH-SY5Y细胞随机分为对照组、OGD组及阿利吉仑低、中和高剂量(5.0、10.0和20.0 μmol/L)组。CCK-8法检测细胞活力;ELISA检测兴奋性氨基酸转运蛋白2(EAAT2/GLT-1)、兴奋性氨基酸转运蛋白3(EAAT3/EAAC1)、兴奋性氨基酸转运蛋白4(EAAT4)、内皮素1(ET-1)和S100钙结合蛋白β亚基(S-100β)的表达;Hoechst 33258染色观察SH-SY5Y细胞形态变化;乳酸(LD)测试盒和超微量Na +-K +-ATP酶测试盒检测LD含量和Na +-K +-ATPase活性。结果: OGD损伤4 h时,细胞相对活力不足60%,因此4 h可作为后续实验OGD造模时间。与对照组相比,OGD组的GLT-1、EAAC1和EAAT4表达显著下调( P <0.05),ET-1和S-100β的表达显著上调( P <0.05);与OGD组相比,阿利吉仑组的GLT-1、EAAC1和EAAT4表达呈剂量依赖性上调, ET-1和S-100β表达呈剂量依赖性下调( P <0.05)。Hochest 33258染色结果表明,阿利吉仑可明显减少OGD引起的SH-SY5Y细胞凋亡。与对照组相比,OGD组的LD含量显著升高( P <0.05),Na +-K +-ATPase活性显著降低( P <0.05);与OGD组相比,阿利吉仑组的LD含量呈剂量依赖性降低,Na +-K +-ATPase活性呈剂量依赖性升高( P <0.05)。结论:阿利吉仑对OGD损伤的SH-SY5Y细胞有一定的保护作用,其机制可能与阿利吉仑上调GLT-1、EAAC1和EAAT4的水平,提高Na +-K +-ATPase的活性,下调ET-1和S-100β的表达及LD含量有关。AIM: To investigate the effects of aliskiren on the injury of SH-SY5Y cells induced by oxygen-glucose deprivation (OGD) and its possible mechanisms. METHODS: The SH-SY5Y cells were randomly divided into control group, OGD group and aliskiren (5.0, 10.0 and 20.0 μmol/L) groups. The cell viability was measured by CCK-8 assay. The levels of excitatory amino acid transporter 2 (EAAT2/GLT-1), EAAT3/EAAC1, EAAT4, endothelin-1 (ET-1) and S100 calcium-binding protein β subunit (S-100β) in the SH-SY5Y cells were detected by ELISA. The morphological changes of the cells were observed by Hoechst 33258 staining. Meanwhile, the content of lactic acid (LD) and activity of Na +-K +-ATPase were also analyzed. RESULTS: The viability of SH-SY5Y cells was not more than 60% after OGD injury for 4 h, so the appropriate time for OGD injury was 4 h. Compared with control group, the protein levels of GLT-1, EAAC1 and EAAT4 in the SH-SY5Y cells of OGD group were significantly decreased ( P <0.05), but the protein levels of ET-1 and S-100β were significantly increased ( P <0.05). Compared with OGD group, treatment with aliskiren dose-dependently increased the protein levels of GLT-1, EAAC1 and EAAT4 in the SH-SY5Y cells, but decreases in the levels of ET-1 and S-100β were observed ( P <0.05). The results of Hochest 33258 staining showed that aliskiren significantly reduced the apoptosis of SH-SY5Y cells. Compared with control group, a significant increase in the content of LD ( P <0.05) and a significant decrease in Na +-K +-ATPase activity ( P <0.05) were found in the SH-SY5Y cells of OGD group. Compared with OGD group, aliskiren dose-dependently decreased the content of LD, but increased the Na +-K +-ATPase activity in the SH-SY5Y cells ( P<0.05 ). CONCLUSION: Aliskiren has good neuroprotective effects on SH-SY5Y cells after OGD injury. The underlying mechanisms may be associated with the increases in the protein levels of GLT-1, EAAC1 and EAAT4, the enhancement of Na +-K +-ATPase activity, and the decreases in the levels of ET-1 a

关 键 词：阿利吉仑 SH-SY5Y细胞 氧糖剥夺 内皮素1 兴奋性氨基酸转运蛋白 

分 类 号：R743.3[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]