<sup>99</sup>Tc <sup>m</sup>-Syt I-C2A突变体蛋白在心肌缺血再灌注诱导的心肌细胞凋亡显像中的应用

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摘要目的探讨⁹⁹Tc^m标记突触结合蛋白I-C2A片段突变体(⁹⁹Tc^m-Syt I-C2A-G4C）在心肌凋亡显像中的应用。方法（1）采用亚锡还原法制备⁹⁹Tc^m-Syt I-C2A-G4C。 ⁹⁹Tc^m-Syt I-C2A-G4C经纯化后通过纸层析法测定放化纯;细胞结合实验测定⁹⁹Tc^m-Syt I-C2A-G4C与磷脂酰丝氨酸的结合能力。（2）选用5头小猪制备心肌缺血-再灌注损伤模型，静脉注射^99mTc-Cyt-C2A-G4C 2 h后进行图像采集，计算心肌异常高放射性区/本底（T/B）比值;同时使用原位末端标记法（TUNEL）分析损伤心肌中细胞凋亡情况。结果（1）标记后的⁹⁹Tc^m-Syt I-C2A-G4C放射化学纯度为（96.42±0.76）%;（2）⁹⁹Tc^m-Syt I-C2A-G4C与喜树碱处理组细胞结合测定的放射性计数是未处理组细胞的5.23±0.38倍;（3） ^99mTc-Cyt-C2A-G4C注射后2 h，心肌部位可见明显的局部放射性异常浓聚，T/B比值为3.40±0.43 ;高放射性心肌组织标本，TUNEL染色亦可见大量黄染的凋亡细胞。结论 ⁹⁹Tc ^m-Syt I-C2A-G4C可用于心肌细胞凋亡显像。

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	钱丽娟
	周俊东
	方纬
	吴锦昌

关键词 ：心肌缺血再灌注, 凋亡显像, 突触结合蛋白I, 锝

Abstract：Objective To investigate application of mutation protein of C2A domain of synaptotagmin I（Syt I-C2A-G4C） labeled with technetium in myocardial apoptosis imaging. Methods （1） The C2A domain of Syt I was labeled with ⁹⁹Tc^m using reduction photoetry of Tin chloride. Radiochemical purity was determined with thin layer chromatography. The binding activity with phosphatidylserine of radiolabeled protein was assessed using cell binding experiment.（2） Myocardial ischemia-reperfusion models were produced in 5 pigs. SPECT images were acquired at 2h after post injection of ^99mTc-Cyt-C2A-G4C via the tail， and the ratio of target/background was also cal- culated using region of interest（ROI） method. Apoptosis in the ischemic myocardium was assessed using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay. Results （1） The radiochemical purity of ⁹⁹Tc^m-Syt I-C2A-G4C was（96.42±0.76）%.（2） The radioactivity in the camptothecin-treated group was （5.23±0.38） folds higher than that of non-treated viable control group.（3）At 2 h post-injection of the radiotracer， the focal uptake of radioactivity was observed at the ischemic myocardium areas， and T/B ratio was 3.40± 0.43 . Myocardial apoptosis was confirmed by TUNEL assay in the ischemic myocardium with high radioactivity. Conclusion ⁹⁹Tc^m-Syt I-C2A-G4C should be a potential radiotracer for imaging of myocardial apoptosis.

Key words： myocardial ischemia-reperfusion apoptosis imaging Synaptotagmin I Technetium

基金资助:

南京医科大学科技发展基金资助项目（NY0714）;苏州市科技局社会发展基金资助项目（SZD0894）和苏州市科技计划兴卫青年科技计划资助项目（SWKQ0818）

引用本文:

钱丽娟1，周俊东1，方纬2，吴锦昌1. ⁹⁹Tc ^m-Syt I-C2A突变体蛋白在心肌缺血再灌注诱导的心肌细胞凋亡显像中的应用[J]. 实用心电学杂志, 2012, 21(2): 82-.

链接本文:

http://zzs.ujs.edu.cn/syxd/CN/ 或 http://zzs.ujs.edu.cn/syxd/CN/Y2012/V21/I2/82

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