6-羟基多巴模型大鼠salsolinol氮甲基转移酶活性检测

发布时间：2018-07-03 01:52

本文选题：Salsolinol氮甲基转移酶 + 帕金森病　；参考：《中国比较医学杂志》2017年12期

【摘要】：目的应用6-羟基多巴(6-hydroxydopamine,6-OHDA)制作帕金森病(Parkinson’s disease,PD)大鼠模型,并对大鼠外周血淋巴细胞中salsolinol氮甲基转移酶(SNMT)的活性进行测定。方法利用单侧双点注射6-OHDA损毁大鼠纹状体,构建PD模型并对模型进行行为学评价;从模型大鼠外周血淋巴细胞中提取SNMT粗酶液,建立酶反应产物N-methyl-salsolinol(NMSal)的高效液相色谱串联三重四极杆质谱检测方法,以NMSal的生成量表征SNMT的活性。结果 18只经过6-OHDA注射的大鼠共有7只经阿朴吗啡诱导后表现为恒定向未损毁侧旋转(7 r/min),建模成功率为38.9%。建立了基于多反应监测技术的SNMT活性的高选择性、高灵敏度、高重复性的表征方法。该方法中NMSal的检出限和定量限分别达到49 pmol/L和98 pmol/L,日内和日间精密度均在6.0%以下。检测结果显示PD组大鼠外周血淋巴细胞中SNMT的活性与假手术组、正常组相比差异有显著性(P0.01,n=5),而正常组和假手术组之间差异无显著性(P0.05,n=5)。结论外周血淋巴细胞中SNMT活性可能反映出PD发病状况,有望成为诊断的指标之一。
[Abstract]:Objective to establish a rat model of Parkinson's disease (PD) with 6-hydroxydopa (6-OHDA) and to determine the activity of salsolinol N-methyltransferase (SNMT) in peripheral blood lymphocytes of rats. Methods the rat striatum was damaged by 6-OHDA injection of 6-OHDA, and the PD model was established, and the model was evaluated by behavioral evaluation, and the crude enzyme solution of SNMT was extracted from the peripheral blood lymphocytes of the model rats. A method for the detection of N-methyl-salsolinol (NMSal) by high performance liquid chromatography (HPLC) tandem triplex quadrupole mass spectrometry (HPLC-MS) was established. The activity of SNMT was characterized by the yield of NMSal. Results of 18 rats injected with 6-OHDA, 7 rats were induced by apomorphine and rotated to the undamaged side (7 r/min). The success rate of modeling was 38.9%. A high selectivity, high sensitivity and high repeatability characterization method of SNMT activity based on multi-reaction monitoring technique was established. The detection limit and quantitative limit of NMSal in this method are 49 pmol / L and 98 pmol / L, respectively. The intra-day and inter-day precision are below 6.0%. The results showed that the activity of SNMT in peripheral blood lymphocytes of PD group was significantly higher than that of sham operation group and normal group (P 0.01), but there was no significant difference between normal group and sham operation group (P 0.05). Conclusion SNMT activity in peripheral blood lymphocytes may reflect the pathogenesis of PD and may be one of the diagnostic indexes.
【作者单位】：中国科学院生态环境研究中心;北京理工大学生命学院;
【分类号】：R-332;R742.5

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