CAJ | 학술논문

目的应用99TcmO-4标记突触结合蛋白I-C2A片段(99Tcm-Syt I-C2A)评价缺血预处理对心肌缺血再灌注损伤的保护作用.方法 (1)采用2-亚氨基噻吩盐酸盐(2-IT)方法标记Syt I-C2A片段,纸层析法测定99Tcm-Syt I-C2A放化纯,用喜树碱处理的Jurket细胞测定标记蛋白质活性.(2)制备心肌缺血再灌注大鼠模型(A组)和心肌缺血预处理大鼠模型(B组)各6只,分别由尾静脉注射99Tcm-syt I-C2A 7.4 MBq,注射后1h处死动物,取出心脏,用生理盐水将心肌冲洗干净,并进行氯化三苯基四氮唑(TTC)染色,根据染色结果,分别取2组缺血损伤心肌和正常心肌,测量质量及放射性计数,比较2组缺血损伤心肌和正常心肌的每克组织百分注射剂量率(%ID/g).采用SPSS 12.0软件行统计分析,数据间比较用t检验.结果 (1)标记后的99Tcm-Syt I-C2A放化纯为(98.90±o.43)%,标记蛋白质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(10.99±O.55)倍.(2)A组缺血损伤心肌摄取99Tcm-Syt I-C2A(2.41±0.32)%ID/g,正常心肌为(O.16±O.02)%ID/g;而B组缺血损伤心肌和正常心肌的放射性摄取则分别为(0.46±0.05)和(0.20±0.05)%ID/g.B组缺血损伤心肌的99Tcm-Syt I.C2A摄取量明显低于A组(t=8.52,P<O.01).结论 99Tcm-Syt I-C2A可用于定量评价缺血预处理抑制心肌细胞凋亡而产生的对缺血再灌注致心肌损伤的保护作用.
목적 응용99TcmO-4표기돌촉결합단백I-C2A편단(99Tcm-Syt I-C2A)평개결혈예처리대심기결혈재관주손상적보호작용.방법 (1)채용2-아안기새분염산염(2-IT)방법표기Syt I-C2A편단,지층석법측정99Tcm-Syt I-C2A방화순,용희수감처리적Jurket세포측정표기단백질활성.(2)제비심기결혈재관주대서모형(A조)화심기결혈예처리대서모형(B조)각6지,분별유미정맥주사99Tcm-syt I-C2A 7.4 MBq,주사후1h처사동물,취출심장,용생리염수장심기충세간정,병진행록화삼분기사담서(TTC)염색,근거염색결과,분별취2조결혈손상심기화정상심기,측량질량급방사성계수,비교2조결혈손상심기화정상심기적매극조직백분주사제량솔(%ID/g).채용SPSS 12.0연건행통계분석,수거간비교용t검험.결과 (1)표기후적99Tcm-Syt I-C2A방화순위(98.90±o.43)%,표기단백질여희수감처리조세포결합측정적방사성계수시미처리조세포적(10.99±O.55)배.(2)A조결혈손상심기섭취99Tcm-Syt I-C2A(2.41±0.32)%ID/g,정상심기위(O.16±O.02)%ID/g;이B조결혈손상심기화정상심기적방사성섭취칙분별위(0.46±0.05)화(0.20±0.05)%ID/g.B조결혈손상심기적99Tcm-Syt I.C2A섭취량명현저우A조(t=8.52,P<O.01).결론 99Tcm-Syt I-C2A가용우정량평개결혈예처리억제심기세포조망이산생적대결혈재관주치심기손상적보호작용.
Objective Precondition is an approach to myocardial protection during ischemia-reper-fusion by inhibiting myocardial cell apoptosis.The purpose of this study was to evaluate the cardioprotective effect using 99Tcm-syuaptotagmin I (Syt I)-C2A to detect myocardial cell apoptosis in the myocardial is-chemia-repedusion rat model.Methods (1) The C2A domain of Syt I was labeled with 99Tcm using 2-iminothiophene hydrochloride (IT) method.Radiochemical purity was determined with thin layer chroma-tography.The binding activity of radiolabeled protein was assessed using eamptothecin-treated Jurket cells.(2) One group of 6 rats was prepared for myocardial ischemia-reperfusion model(A group),and another group of 6 rats was prepared for myocardial ischemia precondition model(B group).99Tcm-Syt I-C2A was injected via the tail vein at a dosage of about 7.4 MBq.At 1h after injection,the rat was sacrificed,and the heart was removed to rinse with saline and dye with triphenyl tetrazolium eoride (TTC).According to the resdt of myocardial dye,theischemic myoeardium was separated from the viable myocardium and weight was measured,and then its radioactivity was determined by gamma counting.The difference of radioactive uptake in the ischemic myocardium between these two group models was compared using percentage activity of injection dose per gram of tissue(%ID/g)±standard deviation(x±s).SPSS 12.0 was used for data analy-sis,and t-test was used to compare data.Results (1) The radiochemical purity of 99Tcm-Syt I-C2A was (98.90±0.43)%,and the radioactivity in the camptothecin-treated group was (10.99±0.55) folds higher than that of non-treated viable control group.(2)In the ischemia-reperfusion model,the radioactive uptake of 99Tcm-Syt,I-C2A was(2.41±0.32)%ID/g in the ischemic myocardium,and(0.16±O.02)%ID/g in the nomud myocardiunm.However,in the myocardial ischemia precondition model,(0.46±0.05)%ID/g in the isehemic myocardium was measured,and(0.20±0.05)%ID/g in the normal myocardium.Uptake of 99Tcm-Syt I-C2A in ischemic myocrdium showed statistically significant difference (t=8.52,P<O.01) between these two groups of models.Conclusion 99Tcm-C2A-glutathione S-transferase (GST) may be used to quantita-tively detect cardioprotective effect of ischemic precondition,which could inhibit myocardium apoptosis on ischemic myocardium in the ischemia-reperfusion rat model.