Study of 99mTc-annexin V uptake in apoptotic cell models of Parkinson's disease
Study of 99mTc-annexin V uptake in apoptotic cell models of Parkinson's disease
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DOI：
10.1097/mnm.0b013e3282f1ac9a
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发表时间：
2007-12-01
影响因子：
1.5
通讯作者：
Zhang, Yongxue
中科院分区：
文献类型：
--
作者：
Cao, Wei;Huang, Jinsha;Zhang, Yongxue
目的探讨放射性标记膜联蛋白V（Annexin V）显像早期诊断帕金森病的可行性。通过向PC 12和SH-SY 5 Y细胞系施用不同浓度的1-甲基-4-苯基吡啶鎓来产生帕金森病的细胞模型。流式细胞仪检测细胞凋亡率。用~（99）Tc-肼基烟酰胺（HYNIC）标记Annexin V。使用帕金森病的细胞模型进行细胞结合研究。结果（TC）-T-99 m-HYNIC-annexin V的比活度为3.7-74 × 10（5）Bq中心点mg（-1）蛋白。Tc-99 m-HYNIC-annexin V与模型细胞的体外结合具有特异性、饱和性和时间依赖性。Scatchard分析得到的Kd为7.16 +/- 1.78 nmol中心点1（-1），B-max值为179 +/- 33 fmol/10（6）个细胞（PC 12）和220 +/- 26 fmol/10（6）个细胞（SHSY 5 Y）。不同浓度的MPP+可剂量依赖性地诱导细胞凋亡;膜结合放射性标记的annexin V活性所指示的细胞摄取99 mTc-HYNIC-annexin V与FITC-annexin V流式细胞术观察到的总荧光呈线性相关结论Tc 99 m-HYNIC-annexin V具有受体结合活性，对帕金森病细胞模型具有较高的亲和力。放射性的摄取与细胞凋亡率相关，因此，Tc-99 m-annexin V是一种潜在的成像剂，用于检测帕金森病的早期神经元损伤。
Objective To investigate the feasibility of radiolabelled annexin V imaging for early diagnosis of Parkinson's disease.Methods Tc-99m-HYNIC-annexin V was prepared and its binding to apoptotic cell models of Parkinson's disease was studied in vitro. Cellular models of Parkinson's disease were produced by administering different concentrations of 1-methyl-4-phenylpyridinium to PC12 and SH-SY5Y cell lines. Cell apoptosis rates were analysed by flow cytometry. Annexin V was labelled with Tc-99m by hydrazinonicotinamide (HYNIC). Cell binding studies were carried out using cellular models of Parkinson's disease. Cell uptake studies were also performed after different levels of MPP+ treatment, and the correlation between the degree of apoptosis and Tc-99m-HYNIC-annexin V uptake was analysed.Results The specific activity of (TC)-T-99m-HYNIC-annexin V was 3.7-74 x 10(5) Bq center dot mg(-1) protein. In-vitro binding of Tc-99m-HYNIC-annexin V to model cells was specific, saturable and time dependent. Scatchard analysis gave a K-d of 7.16 +/- 1.78 nmol center dot l(-1), B-max values of 179 +/- 33 fmol per 10(6) cells (PC12) and 220 +/- 26 fmol per 10(6) cells (SHSY5Y). MPP+ at different concentrations can induce cell apoptosis in a dose-dependent manner; cellular uptake of 99mTc-HYNIC-annexin V as indicated by membrane-bound radiolabelled annexin V activity was linearly correlated with total fluorescence, as observed by FITC-annexin V flow cytometry (PC12: r = 0.924; SH-SY5Y: r = 0.937, P < 0.01).Conclusions Tc-99m-HYNIC-annexin V retains its receptor-binding activity and has a high affinity to cellular models of Parkinson's disease. The uptake of radioactivity correlated well with cell apoptosis rates; thus, Tc-99m-annexin V is a potential imaging agent with which to detect early neuron damage in Parkinson's disease.