摘要
目的探讨大麻素Ⅱ型受体(CB2R)激活对1-甲基-4-苯基-吡啶离子(MPP+)诱导的SH-SY5Y细胞损伤的保护作用机制。方法培养SH-SY5Y细胞,根据药物处理不同将其分为对照组、MPP+组、CB2R激动剂JWH133+MPP+组、CB2R抑制剂AM630+JWH133+MPP+组。应用激光扫描共聚焦显微镜分析技术检测SH-SY5Y细胞铁水平变化,应用免疫印迹法(Western blot)检测各组细胞二价金属离子转运蛋白1(DMT1)的表达。结果与对照组相比,MPP+组细胞荧光强度明显降低,差异有显著性(F=26.620,q=10.410,P<0.05);用JWH133预处理后,JWH133+MPP+组细胞荧光淬灭程度低于MPP+组,差异有显著性(q=4.868,P<0.05);而且应用AM630可以逆转JWH133的这种作用,AM630+JWH133+MPP+组和JWH133+MPP+组荧光强度相比差异具有显著性(q=5.619,P<0.05)。与对照组相比,MPP+组细胞的DMT1蛋白表达量显著升高(F=9.493,q=4.109,P<0.05);JWH133+MPP+组细胞DMT1蛋白表达量下降,与MPP+组相比差异具有显著性(q=4.771,P<0.05);而AM630可阻断JWH133的作用,AM630+JWH133+MPP+组细胞DMT1蛋白表达量较JWH133+MPP+组显著升高(q=6.240,P<0.05)。结论激活CB2R可以抑制MPP+对DMT1蛋白表达的诱导从而调节SH-SY5Y细胞铁水平降低。
Objective To investigate the protective mechanism of cannabinoidⅡtype receptor(CB2R)activation against 1-methyl-4-phenylpyridine(MPP+)-induced SH-SY5Y cell injury.Methods SH-SY5Y cells were cultured and divided into control group,MPP+group,JWH133+MPP+group,and AM630+JWH133+MPP+group according to the drug treatment me-thod.Laser scanning confocal microscopy was used to observe the change in iron level in SH-SY5Y cells,and Western blot was used to measure the expression of divalent metal ion transporter 1(DMT1)in each group.Results Compared with the control group,the MPP+group had a significant reduction in the fluorescence intensity of cells(F=26.620,q=10.410,P<0.05).After pretreatment with the CB2R agonist JWH133,the JWH133+MPP+group had a significantly lower fluorescence quenching degree than the MPP+group(q=4.868,P<0.05).The effect of JWH133 could be reversed by the CB2R inhibitor AM630,and there was a signi-ficant difference in fluorescence intensity between the AM630+JWH133+MPP+group and the JWH133+MPP+group(q=5.619,P<0.05).Compared with the control group,the MPP+group had a significant increase in the protein expression level of DMT1(F=9.493,q=4.109,P<0.05),while the JWH133+MPP+group had a significant reduction in the protein expression level of DMT1(q=4.771,P<0.05).AM630 could block the effect of JWH133,and compared with the JWH133+MPP+group,the AM630+JWH133+MPP+group had a significant increase in the protein expression level of DMT1(q=6.240,P<0.05).Conclusion Activation of CB2R can inhibit the protein expression of DMT1 induced by MPP+and thus regulate the reduction in iron level in SH-SY5Y cells.
作者
潘东
马泽刚
PAN Dong;MA Zegang(Department of Physiology,School of Basic Medicine,Qingdao University,Qingdao 266071,China)
出处
《青岛大学学报（医学版）》
CAS
2021年第2期206-209,共4页
Journal of Qingdao University(Medical Sciences)
基金
山东省重点研发计划资助项目(2019GSF108095)。
