论文部分内容阅读
目的研究帕金森病(PD)的发病机制,提高临床的正确诊断及其探索基因治疗。方法和结果MTT法、末端标记法(TUNEL)、流式细胞仪(FCM)、电镜、DNA电泳、RT-PCR和Western blot方法检测显示MPP~+致使MES23.5细胞活力显著减低,线粒体膜电势(△m)下降和氧自由基增加,MPTP、MPP~+和(或)6-OHDA使黑质细胞和(或)PC12细胞发生凋亡,Caspase-3mRNA表达增加。微透析与HPLC、免疫组化等显示PD患者血IgG、脂多糖(LPS)、Fe~(2+)致使大鼠黑质酪氨酸羟化酶(TH)阳性细胞减少,纹状体多巴胺(DA)降低,MPTP致使猴纹状体谷氨酸、天门冬氨酸、一氧化氮增高。~(125)I-β-CIT、~(99m)Tc-TRODAT-1及~(125)I-IBZM、~(131)I-AIBZM单光子发射断层扫描和(或)放射自显影显示PD猴、大鼠或小鼠模型的纹状体DA转运载体(DAT)功能降低和D_2DA受体活性增强。转TH基因成肌细胞或转TH与GTP环水解酶-1(GCH)成纤维细胞,以及腺病毒介导的胶质源性神经营养因子(GDNF)移植治疗显示PD大鼠模型的旋转行为改善,TH和GDNF基因表达,以及DA浓度增加。结论氧化应激、兴奋性毒性、免疫异常等与PD发病有关。DAT和D_2DA受体显像有助于提高疾病的诊断和病情监测。TH和(或)GDNF基因可能是PD治疗的有效措施。
Objective To study the pathogenesis of Parkinson’s disease (PD), improve clinical diagnosis and explore gene therapy. Methods and Results MTT assay, TUNEL assay, flow cytometry (FCM), electron microscopy, DNA electrophoresis, RT-PCR and Western blot showed that MPP ~ + could significantly reduce the viability of MES23.5 cells, mitochondrial membrane potential (△ m) decreased and oxygen free radicals increased. MPTP, MPP ~ + and / or 6-OHDA induced apoptosis in the substantia nigra and / or PC12 cells and increased expression of Caspase-3 mRNA. Microdialysis, HPLC and immunohistochemistry showed that the levels of tyrosine hydroxylase (TH) positive cells in substantia nigra were reduced in blood serum and lipopolysaccharide (LPS) DA) decreased, MPTP led to the monkey striatum glutamate, aspartate, nitric oxide increased. PD-1 (125) I-β-CIT, 99m Tc-TRODAT-1 and 125 I-IBZM and 131I-AIBZM single photon emission tomography and / or autoradiography , Decreased function of the striatum DA transporter (DAT) and increased D 2 DA receptor activity in rat or mouse models. Transplantation of TH-derived myoblasts or TH and GTP-cyclo-hydrolase-1 (GCH) fibroblasts, and adenovirus-mediated glial derived neurotrophic factor (GDNF) transplantation showed an improvement in the rotational behavior of the PD rat model , TH and GDNF gene expression, and increased DA concentration. Conclusion Oxidative stress, excitotoxicity and immune abnormalities are related to the pathogenesis of PD. DAT and D_2DA receptor imaging help to improve disease diagnosis and disease surveillance. TH and / or GDNF genes may be effective measures of PD treatment.