CAJ | 학술논문

目的制备99Tcm-联肼尼克酰胺(HYNIC)-膜联蛋白(Annexin)B1并对其探测细胞凋亡的潜力进行生物学评价.方法应用99Tcm间接标记蛋白质的方法,以HYNIC作为双功能螯合剂制备99Tcm-HYNIC-Annexin B1.采用与活化人血小板结合实验检测99Tcm-HYNIC-Annexin B1的体外生物活性.采用地塞米松诱导小鼠胸腺凋亡的动物模型和anti-Fas单克隆抗体诱导小鼠肝凋亡的动物模型(均设相应对照组),检测99Tcm-HYNIC-Annexin B1探测体内细胞凋亡的潜力,并用原位末端标记法(TUNEL)染色证实细胞凋亡.采用配对t检验进行统计学处理.结果间接标记法制备的99Tcm-HYNIC-Annexin B1具有很高的放化纯(＞96%)和较好的体外稳定性.与活化人血小板的结合实验表明,99Tcm-HYNIC-Annexin B1具有与磷脂酰丝氨酸(PS)残基结合的生物活性.地塞米松诱导18 h后,小鼠胸腺对99Tcm-HYNIC-Annexin B1的摄取为对照组的3.50倍(t=5.234,P＜0.01).anti-Fas诱导后2 h时,小鼠肝对99Tcm-HYNIC-Annexin B1的摄取为对照组的2.02倍(t=6.178,P＜0.01).结论采用99Tcm间接标记法制备的99Tcm-HYNIC-Annexin B1具有很高的放化纯及较好的体外稳定性.99Tcm-HYNIC-Annexin B1具有与PS结合的体外生物活性和探测体内细胞凋亡的潜力,是一种新的细胞凋亡显像剂.
목적 제비99Tcm-련정니극선알(HYNIC)-막련단백(Annexin)B1병대기탐측세포조망적잠력진행생물학평개.방법 응용99Tcm간접표기단백질적방법,이HYNIC작위쌍공능오합제제비99Tcm-HYNIC-Annexin B1.채용여활화인혈소판결합실험검측99Tcm-HYNIC-Annexin B1적체외생물활성.채용지새미송유도소서흉선조망적동물모형화anti-Fas단극륭항체유도소서간조망적동물모형(균설상응대조조),검측99Tcm-HYNIC-Annexin B1탐측체내세포조망적잠력,병용원위말단표기법(TUNEL)염색증실세포조망.채용배대t검험진행통계학처리.결과 간접표기법제비적99Tcm-HYNIC-Annexin B1구유흔고적방화순(＞96%)화교호적체외은정성.여활화인혈소판적결합실험표명,99Tcm-HYNIC-Annexin B1구유여린지선사안산(PS)잔기결합적생물활성.지새미송유도18 h후,소서흉선대99Tcm-HYNIC-Annexin B1적섭취위대조조적3.50배(t=5.234,P＜0.01).anti-Fas유도후2 h시,소서간대99Tcm-HYNIC-Annexin B1적섭취위대조조적2.02배(t=6.178,P＜0.01).결론 채용99Tcm간접표기법제비적99Tcm-HYNIC-Annexin B1구유흔고적방화순급교호적체외은정성.99Tcm-HYNIC-Annexin B1구유여PS결합적체외생물활성화탐측체내세포조망적잠력,시일충신적세포조망현상제.
Objective Annexin B1,a novel Ca2+ -dependent phosphatidylserine(PS)-binding protein,has been shown to have a high affinity for PS exposed on the surface of apoptotic cells or activated platelets.The aim of this study was to develop and bioevaluate an Annexin B1 based PS-targeting radiotracer labeled with 99Tcm.Methods Annexin B1 was indirectly labeled with 99Tcm using hydrazinonicotinamide(HYNIC)as a bifunctional chelator agent.Binding assay with human activated platelets was used to evaluate the biological activity of 99Tcm-HYNIC-Annexin B1 in vitro.The potential of 99Tcm-HYNIC-Annexin B1 to detect apoptosis in vivo was evaluated in mice models with dexamethasone-inducecd thymus apoptosis and anti-Fas antibody induced liver apoptosis.The paired t-test was used to analyse the data.Results The labeling procedure yielded a compound with radiochemical purity higher than 96% and good in vitro stability.Plate lets binding assay indicated that 99Tcm-HYNIC-Annexin B1 retained their PS binding activity in vitro.The per-centage activity of injection dose per gram of tissue(%ID/g)of mouse thymus showed a 3.50-fold increase at 18 h after administration of dexamethasone compared with control mice(t=5.234,P＜0.01).Radionuclide imaging showed a markedly increased uptake of 99Tcm-HYNIC-Annexin B1 in the liver of anti-Fas antibody treated mice.The% ID/g of apoptotic murine liver showed a 2.02-fold increase at 2 h after the administration of anti-Fas antibody compared with control mice(t=6.178,P＜0.01).Conclusions These data suggest that 99Tcm-HYNIC-Annexin B1 can be prepared with high radiochemical purity and in vitro stability.These data also suggest that 99Tcm-HYNIC-Annexin B1 retains its in vitro and in vivo biological activities.It may therefore be useful as a novel radioligand for the noninvasive imaging of PS externalization associated with apoptosis.