右美托咪定通过激活AMPK减轻TNF-α诱导人成神经细胞瘤细胞系SH-SY5Y损伤

Dexmedetomidine alleviates TNF-α-inducedinjury of human neuroblastoma cell line SH-SY5Y through activating AMPK

目的 研究右美托咪定(DEX)对TNF-α诱导的人成神经细胞瘤细胞系的作用及其机制。方法 采用CCK8检测细胞活性,确定最佳DEX和TNF-α剂量,细胞分为:对照组(control组)、模型组(model组)、DEX干预模型组(DEX组)、DEX联合compound C干预模型组(DEX+CC组);Western blot检测细胞中p-AMPK、SNHP、KIF5B、Drp1、OPA1蛋白表达,ELISA检测IL-1β、IL-6水平,相应试剂盒测定线粒体膜电位(Δψm)、呼吸链复合酶活性(complexⅠ-Ⅳ)、ATP、MDA、SOD、GSH、ROS。结果 模型组较对照组p-AMPK活性、OPA1及SNPH水平显著降低,但Drp1和KIF5B水平显著增高(P<0.01),DEX组较model组complexⅠ~Ⅳ及Δψm、ATP、GSH、SOD水平显著增高,而MDA、IL-1β、IL-6水平显著降低(P<0.01);DEX+CC组较DEX组complexⅠ~Ⅳ及Δψm、ATP、GSH、SOD水平显著降低,而MDA、IL-1β、IL-6水平显著增高(P<0.01)。结论 DEX通过依赖AMPK的方式改善线粒体功能、减少氧化应激及炎性反应,减轻TNF-α诱导的细胞损伤。

Objective To study the effect of dexmedetomidine(DEX)on TNF-α-induced injury of human neuroblastoma cell line SH-SY5Y and its mechanism.Methods CCK8 assay was used to detect cell activity and determine the optimal dose of DEX and TNF-α.The cells were divided into control group,model group,DEX intervention model group,and the intervention group of DEX plus compound C.Western blot was used to detect the expression of p-AMPK,SNHP,KIF5B,Drp1 and OPA1,and ELISA was used to detect the level of IL-1βand IL-6.Mitochondrial membrane potential(Δψm),respiratory chain complex enzyme activity(complexⅠ-Ⅳ),ATP,MDA,SOD,GSH,ROS were determined with commercially available kits.Results Compared with control group,level of p-AMPK,OPA1 and SNPH l in model group significantly decreased,while the level of Drp1 and KIF5B significantly increased(P<0.01).The level of complexⅠ~Ⅳ,Δψm,ATP,GSH and SOD in DEX group significantly increased as compared with model group.The level of MDA,IL-1βand IL-6 significantly decreased(P<0.01).Compared with DEX group,the level of ComplexⅠ-Ⅳ,Δψm,ATP,GSH and SOD in DEX+CC group significantly decreased,while the level of MDA,IL-1βand IL-6 significantly increased(P<0.01).Conclusions DEX improves mitochondrial function,alleviates oxidative stress and inflammatory response as well as TNF-α-induced cell damage with an AMPK dependent mechanism.