摘要
目的研究亚硝基化的二硫键异构酶(PDI)对甲基苯丙胺(METH)致小鼠海马及纹状体区α-突触核蛋白(α-SN)表达的影响。方法利用C57小鼠METH亚急性中毒模型,使用一氧化氮合酶(NOS)抑制剂N-硝基-L-精氨酸(L-NNA)与METH共同造模,实验分为对照组、L-NNA组、METH组、METH+L-NNA组。Western Blotting技术检测各组海马及纹状体区内一氧化氮合酶(NOS)、PDI及其S亚硝基化(PDI-SNO)和α-SN表达情况。一氧化氮试剂盒检测各组一氧化氮含量。结果 METH给药后NOS、一氧化氮含量、PDI-SNO、α-SN表达较对照组均明显升高(P<0.05);L-NNA与METH共处理后与METH组对比,NOS表达下降(P<0.05),一氧化氮含量明显减少(P<0.05),能够显著抑制PDI-SNO表达(P<0.05),伴随α-SN表达降低(P<0.05)。而MET H+L-NNA组、L-NNA组与对照组比较均无明显差异(P>0.05)。结论 METH作用后诱导NOS活化,一氧化氮含量升高,PDI发生显著S亚硝基化而功能失活,导致相关脑区α-SN表达升高,而NOS抑制剂L-NNA能部分缓解METH神经毒性作用。
Objective To investigate the role of S-nitrosylation of protein disulphide isomerasec in methamphetamine（METH）-induced expression of alpha-synuclein（α-SN） in mouse hippocampus and striatum neurons.Methods Forty C57 BL/6 mice were randomized equally into saline control group,METH group,L-NNA（a NOS inhibitor） group and L-NNA plus METH group.All the agents were injected intraperitoneally at an interval of 12 h,and a total of 8 injections were administered;in LNNA plus METH group,METH was injected 30 min after L-NNA in each treatment.Western Blotting was used to detect the expression of nitric oxide synthase（NOS）,α-SN,PDI and S-nitrosylation of protein disulphide isomerase（PDI-SNO） in the hippocampus and striatum of the mice,and nitric oxide（NO） levels were determined using a NO assay kit.Results In METH group,the levels of NOS,PDI-SNO,α-SN and NO all increased significantly compared with those in the control group（P〈0.05）.Combined treatment with L-NNA and METH,compared with METH alone,resulted in significantly lowered expression of NOS,NO,PDI-SNO and α-SN in the hippocampus and striatum of the mice（all P〈0.05）.No significant differences were found in NOS,NO,PDI-SNO or α-SN expressions among METH ＋ L-NNA,L-NNA and control groups（P〈0.05）.Conclusion METH induces the activation of NOS and increases NO level to cause the occurrence of PDI-SNO,leading subsequently to increased expression of α-SN in mouse striatum and hippocampus.L-NNA,the inhibitor of NOS,can partly relieve nervous system toxicity induced by METH.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2017年第10期1386-1390,共5页
Journal of Southern Medical University
基金
国家自然科学基金(81373240)
广东省自然科学基金(2014A030313300
2014A030310025)~~
