~（188）Re标记药物血管内给药对兔VX_2肝肿瘤模型的治疗前研究
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摘要
第一部分兔VX2肝肿瘤模型的建立及CT、DSA表现
目的利用CT定位经皮穿刺直接推送瘤块法建立兔VX2移植性肝肿瘤模型,并研究VX2移植性肝肿瘤的CT、DSA影像学特征。
材料与方法136只新西兰大白兔,采用CT定位下经皮穿刺肝脏后推送VX2瘤块法制作肝肿瘤模型,计算兔种植3周后死亡率、成瘤率,分析VX2移植性肝肿瘤的CT、DSA影像表现,选取CT证实的成瘤兔5只,观察其肝脏大体及光镜下瘤组织形态特征。结果兔种植后3周内死亡21只,死亡率16.7%(21/136),肝脏肿瘤生长102只,成瘤率75.0% ( 102/136)。CT、DSA显示VX2肝肿瘤具有丰富的肝动脉血供。5只成瘤兔都得到病理学证实。
结论CT定位下经皮穿刺直接推送瘤块法是建立兔VX2肝肿瘤模型的一种新方法;该模型肿瘤具有丰富的肝动脉血供,适合用于血管内治疗的实验研究。
第二部分188Re标记碘化油、聚合人白蛋白、锡硫胶体的制备
目的探讨188Re-碘化油、聚合人白蛋白(MAA)、锡硫胶体(SnSC)的标记方法。
方法①在温度为75~120℃和不同实验条件下,直接标记法制备188Re-碘化油,观测188Re-碘化油标记率在室温和37℃人血浆中随时间的变化情况;②市售MAA药盒(临床用于99Tcm标记)中加入SnC12·2H2O后进行188Re标记聚合人白蛋白实验,测定不同时间点标记产物的标记率;③改变加入硫胶体药盒(SC kit)SnC12·2H2O和Na2S2O3·5H2O用量后进行188Re标记SC(Sn)实验,测定不同时间标记物的标记率及其体外稳定性。
结果①直接法制备188Re-碘化油,在120℃的最佳标记条件下,标记物标记率可达(99.11±0.22)%;标记物在室温和37℃人血浆中放置72h,标记率可保持在(90.91±1.93)%和(92.82±1.22)%。②MAA药盒在加入500μL SnCl2(20mg/mL)混匀后,188Re-MAA的标记率为(99.64±0.09)%,72h以后,血清中188Re-MAA的标记率为(92.53±1.20)%。③SC kit在加SnC12·2H2O (20mg/ml) 0.5ml,Na2S2O3(8mg/ml) 0.25ml的条件下,188Re标记锡硫胶体标记率可达(98.39±2.66)% ,室温下在锡硫胶体悬液:生理盐水=1:10的溶液中放置72h标记率为(93.23±0.64)%。
结论188Re-碘化油、聚合人白蛋白、锡硫胶体的标记方法简便,标记率高,体外稳定性好。有望用于实体肿瘤的治疗。
第三部分188Re标记药物血管内给药及其兔VX2模型体内的生物学分布
目的研究188Re标记药物经肝动脉给药后在兔VX2肝肿瘤模型体内的生物学分布特征。
材料与方法VX2肝肿瘤模型兔64只,采用经肝动脉途径灌注药物,按注入药物分成5组:碘化油组(A组)、游离188Re组(B组)、188Re-碘化油组(C组)、188Re-白蛋白组(D组)、188Re-锡硫胶体组(E组)。给药后1h、24h B组、C组、D组和E组行SPECT检查,并在规定时间点将兔处死,获取脑、心、肺、肝、脾、肾、肠、肌肉、血液和肿瘤10个组织或器官样本,称重并测定放射性,计算每克组织百分注射剂量(%ID/g)和肿瘤与肝、肾、肌肉、血液、肺的放射性摄取比值。各组每只VX2兔给药前和给药后1h、24h行肝肾功能检查。
结果35只VX2兔接受血管内给药并完成了实验。除B组外,C组(n=6)、D组(n=9)、E组(n=8)VX2兔在给药后1h、24h SPECT显像可见肿瘤内放射性核素浓聚。188Re在VX2兔体内的分布以肿瘤最高,其次为肝、肾,在肺、血、脾分布较低,在肠、心,脑、肌肉中的分布最低,并随时间延长而降低。肿瘤中的放射性摄取明显高于其它组织。比较各组的T/NT(肿瘤/肝、肿瘤/肾、肿瘤/肺、肿瘤/血液、肿瘤/肌肉)放射性比值,B、C、D、E各组之间均存在统计学差异(p<0.05)。在不同时间点间,C组肿瘤/肝比值无显著性差异(p >0.05),D、E组存在统计学差异(p<0.05)。各组给药后丙氨酸氨基转移酶(ALT)和谷草转氨酶(AST)均在第24h升至最高,B组升高幅度较小,A、C、D、E组间,无统计学差异(p>0.05),B组与A、C、D、E组均存在统计学差异(p<0.05)。不同时间点间两两比较皆存在统计学差异。各组给药前后血清尿素(UREA)均无明显变化。
结论采用经导管肝动脉途径给药,188Re-碘化油、188Re-白蛋白、188Re-锡硫胶体均能在VX2兔肿瘤内积聚,且具有较好的滞留性。它们有可能成为内照射治疗肝癌的新药。
PartⅠEstablishment of a Rabbit Model Bearing VX2 Liver Tumor and its CT、DSA Manifestation
Objective To establish a rabbit model of VX2 liver tumor for experimental study, with pieces of VX2 tumor tissue implanted into the left lobe of the rabbit liver via percutaneous puncture under the guide of CT. and to study the CT、DSA imagining features of the liver tumor.
Methods 136 New Zealand white rabbits were established models of VX2 liver tumor via percutaneous puncture under the guide of CT . The death rate of rabbit and the success rate of establishing VX2 liver tumor model was counted respectively. The CT、DSA imagining manifestation of the VX2 liver tumor was analyzed. 5 rabbit models bearing VX2 liver tumor were selected to observe the biological features of tumor by histopathology and microscope photographs.
Results Within three weeks after implantation, 21 rabbits were died, The death rate was 16.7%(21/136), and the success rate of establishing VX2 liver tumor model was 75.0% ( 102/136), except for 13 rabbits without liver tumor. The CT、DSA imagining of tumor indicated that the tumor had liberal blood supply of hepatic artery. The liver tumor of 5 rabbit models were confirmed by pathology.
Conclusion It’s a new method to establishing a rabbit model of VX2 liver tumor with pieces of VX2 tumor tissue implanted into the left lobe of the rabbit liver via percutaneous puncture under the guide of CT. The CT、DSA imagining show that the tumor is hypervascular. The rabbit model bearing VX2 liver tumor is more suitable for endovascular interventional experimental study.
PartⅡThe preparation of 188Re labelled Lipiodol, MAA, stannous sulfur colloid
Objective To study the methods for labeling lipiodol, macroaggregated albumin(MAA), and stannous sulfur colloid with rhenium-188.
Methods①Lipiodol was labeled with 188Re directly at the temperatures from 75℃to 120℃with different reaction conditions. The time-variation labeling efficiency was measured to evaluate the stability of 188Re-lipiodol.②Macroaggregated albumin(MAA)was labeled with 188Re after extra adding stannous chloride into commercial MAA kit, The time-variation labeling efficiency was measured to evaluate the stability of 188Re-MAA.③Different amount of stannous chloride and sodium thioslphate were respectively added into commercial sulfur colloid (SC). Their labeling efficiencies and the in vitro stabilities were tested.
Results①The optimal condition was obtained with the labeling efficiency over 99.11±0.22% at 120℃. More than 90.9±1.9% and 92.8±1.2% of radioactivity was kept in 188Re-lipidol form in room temperature and in 37℃human plasma for 72 hours;②At the condition of additional dosage of 0.5ml stannous chloride(20mg/ml), the labeling efficiency of 188Re-MAA reached(99.64±0.09)%, and(92.53±1.20)% of labeling efficiency was kept in human plasma for 72 hours③At the condition of additional dosage of 0.5ml stannous chloride(20mg/ml) and 0.25ml sodium thioslphate (8mg/ml), the labeling efficiency of 188Re-SC(Sn) reached(98.39±2.66)%, and labeling efficiency was (93.62±0.64)% in the solution (suspending SC to saline=1:10) in room temperature at 72h after preparation.
Conclusion The methods for labeling lipiodol, macroaggregated albumin(MAA), and stannous sulfur colloid with rhenium-188 are convenient,and stable in vitro with higher labeling efficiency, which can potentially be the new radiopharmaceuticals used for the treatment of stereo tumor.
PartⅢThe Biodistribution after Endovascular Injection of Rhenium-188 Labeled Drugs in Rabbit VX2 Liver Tumor Model
Objective To investigate the biodistribution property of rhenium-188 labeled drug after transcatheter arterial injection of rhenium-188 labeled lipiodol, rhenium-188 labeled macroaggregated albumin(MAA), and rhenium-188 labeled stannous sulfur colloid in rabbit VX2 liver tumor model.
Methods 64 rabbits bearing liver tumor were performed transcatheter hepatic arterial injection of lipiodol (group A), Na188ReO4 solution (group B), rhenium-188 labeled lipiodol (group C), rhenium-188 labeled MAA (group D),and rhenium-188 labeled stannous sulfur colloid(group E), respectively. The rabbits in group B、C、D and E were accepted SPECT, then sacrificed at the time of 1h or 24h after administration . Samples of brain, heart, lung, liver, spleen, kidneys, intestine, muscle, blood, and tumor were taken, weighed and measured for radioactivity. The radioactivity of tissue was expressed as percentage of the injected dose per gram of wet tissue weigh(%ID/g). The ratios of tumor to liver, kidneys, muscle, blood, lung were calculated. The ALT、AST and UREA of each of rabbit were detected before and at the time of 1h、24h after administration, respectively.
Results 35 rabbits were accepted endovascular administration, and accomplished in the study. The SPECT imaging of 23 rabbits (group C, n=6; group D, n=9; group E, n=8) shew that radioactivity was accumulated in tumor at the time of 1h or 24h after administration. The biodistribution of rhenium-188 labeled drug shew that higher radioactivities uptake were in tumor, liver and kidneys, lower radioactivities uptake were in intestine, heart, brain and muscle. and radioactivities uptake were moderate in lung, blood, and spleen. Radioactivities were decreased as the time passed. The uptake of radioactivity in tumor was higher than in the other tissues. The ratios of T/NT (tumor/ liver, tumor/ kidney, tumor/ lung, tumor/ blood, and tumor/muscle) in each group were compared. There were statistics differences among group B、C、D and E(p<0.05)each other. Except for group C, the differences of the ratio of tumor/ liver were significant between the time of 1h and 24h after administration in group D and E(p<0.05). In each group, the levels of ALT and AST even rose to the highest at the time of 24h after administration, and the amplitude in group B was the smallest. There were no statistics differences among group A、C、D and E(p>0.05), and the differences were significant between group B and group A、C、D、E(p<0.05), respectively. There were statistics differences between the both of the different time, too. In each group, the levels of UREA weren’t obviously changed.
Conclusion After transcatheter arterial administration, rhenium-188 labeled lipiodol, rhenium-188 labeled MAA, rhenium-188 labeled stannous sulfur colloid has better tumour affinity and retainable accumulation characteristics in tumor which can potentially be the new radioactive agents used for endovascular treatment of liver cancer.
目的利用CT定位经皮穿刺直接推送瘤块法建立兔VX2移植性肝肿瘤模型,并研究VX2移植性肝肿瘤的CT、DSA影像学特征。
材料与方法136只新西兰大白兔,采用CT定位下经皮穿刺肝脏后推送VX2瘤块法制作肝肿瘤模型,计算兔种植3周后死亡率、成瘤率,分析VX2移植性肝肿瘤的CT、DSA影像表现,选取CT证实的成瘤兔5只,观察其肝脏大体及光镜下瘤组织形态特征。结果兔种植后3周内死亡21只,死亡率16.7%(21/136),肝脏肿瘤生长102只,成瘤率75.0% ( 102/136)。CT、DSA显示VX2肝肿瘤具有丰富的肝动脉血供。5只成瘤兔都得到病理学证实。
结论CT定位下经皮穿刺直接推送瘤块法是建立兔VX2肝肿瘤模型的一种新方法;该模型肿瘤具有丰富的肝动脉血供,适合用于血管内治疗的实验研究。
第二部分188Re标记碘化油、聚合人白蛋白、锡硫胶体的制备
目的探讨188Re-碘化油、聚合人白蛋白(MAA)、锡硫胶体(SnSC)的标记方法。
方法①在温度为75~120℃和不同实验条件下,直接标记法制备188Re-碘化油,观测188Re-碘化油标记率在室温和37℃人血浆中随时间的变化情况;②市售MAA药盒(临床用于99Tcm标记)中加入SnC12·2H2O后进行188Re标记聚合人白蛋白实验,测定不同时间点标记产物的标记率;③改变加入硫胶体药盒(SC kit)SnC12·2H2O和Na2S2O3·5H2O用量后进行188Re标记SC(Sn)实验,测定不同时间标记物的标记率及其体外稳定性。
结果①直接法制备188Re-碘化油,在120℃的最佳标记条件下,标记物标记率可达(99.11±0.22)%;标记物在室温和37℃人血浆中放置72h,标记率可保持在(90.91±1.93)%和(92.82±1.22)%。②MAA药盒在加入500μL SnCl2(20mg/mL)混匀后,188Re-MAA的标记率为(99.64±0.09)%,72h以后,血清中188Re-MAA的标记率为(92.53±1.20)%。③SC kit在加SnC12·2H2O (20mg/ml) 0.5ml,Na2S2O3(8mg/ml) 0.25ml的条件下,188Re标记锡硫胶体标记率可达(98.39±2.66)% ,室温下在锡硫胶体悬液:生理盐水=1:10的溶液中放置72h标记率为(93.23±0.64)%。
结论188Re-碘化油、聚合人白蛋白、锡硫胶体的标记方法简便,标记率高,体外稳定性好。有望用于实体肿瘤的治疗。
第三部分188Re标记药物血管内给药及其兔VX2模型体内的生物学分布
目的研究188Re标记药物经肝动脉给药后在兔VX2肝肿瘤模型体内的生物学分布特征。
材料与方法VX2肝肿瘤模型兔64只,采用经肝动脉途径灌注药物,按注入药物分成5组:碘化油组(A组)、游离188Re组(B组)、188Re-碘化油组(C组)、188Re-白蛋白组(D组)、188Re-锡硫胶体组(E组)。给药后1h、24h B组、C组、D组和E组行SPECT检查,并在规定时间点将兔处死,获取脑、心、肺、肝、脾、肾、肠、肌肉、血液和肿瘤10个组织或器官样本,称重并测定放射性,计算每克组织百分注射剂量(%ID/g)和肿瘤与肝、肾、肌肉、血液、肺的放射性摄取比值。各组每只VX2兔给药前和给药后1h、24h行肝肾功能检查。
结果35只VX2兔接受血管内给药并完成了实验。除B组外,C组(n=6)、D组(n=9)、E组(n=8)VX2兔在给药后1h、24h SPECT显像可见肿瘤内放射性核素浓聚。188Re在VX2兔体内的分布以肿瘤最高,其次为肝、肾,在肺、血、脾分布较低,在肠、心,脑、肌肉中的分布最低,并随时间延长而降低。肿瘤中的放射性摄取明显高于其它组织。比较各组的T/NT(肿瘤/肝、肿瘤/肾、肿瘤/肺、肿瘤/血液、肿瘤/肌肉)放射性比值,B、C、D、E各组之间均存在统计学差异(p<0.05)。在不同时间点间,C组肿瘤/肝比值无显著性差异(p >0.05),D、E组存在统计学差异(p<0.05)。各组给药后丙氨酸氨基转移酶(ALT)和谷草转氨酶(AST)均在第24h升至最高,B组升高幅度较小,A、C、D、E组间,无统计学差异(p>0.05),B组与A、C、D、E组均存在统计学差异(p<0.05)。不同时间点间两两比较皆存在统计学差异。各组给药前后血清尿素(UREA)均无明显变化。
结论采用经导管肝动脉途径给药,188Re-碘化油、188Re-白蛋白、188Re-锡硫胶体均能在VX2兔肿瘤内积聚,且具有较好的滞留性。它们有可能成为内照射治疗肝癌的新药。
PartⅠEstablishment of a Rabbit Model Bearing VX2 Liver Tumor and its CT、DSA Manifestation
Objective To establish a rabbit model of VX2 liver tumor for experimental study, with pieces of VX2 tumor tissue implanted into the left lobe of the rabbit liver via percutaneous puncture under the guide of CT. and to study the CT、DSA imagining features of the liver tumor.
Methods 136 New Zealand white rabbits were established models of VX2 liver tumor via percutaneous puncture under the guide of CT . The death rate of rabbit and the success rate of establishing VX2 liver tumor model was counted respectively. The CT、DSA imagining manifestation of the VX2 liver tumor was analyzed. 5 rabbit models bearing VX2 liver tumor were selected to observe the biological features of tumor by histopathology and microscope photographs.
Results Within three weeks after implantation, 21 rabbits were died, The death rate was 16.7%(21/136), and the success rate of establishing VX2 liver tumor model was 75.0% ( 102/136), except for 13 rabbits without liver tumor. The CT、DSA imagining of tumor indicated that the tumor had liberal blood supply of hepatic artery. The liver tumor of 5 rabbit models were confirmed by pathology.
Conclusion It’s a new method to establishing a rabbit model of VX2 liver tumor with pieces of VX2 tumor tissue implanted into the left lobe of the rabbit liver via percutaneous puncture under the guide of CT. The CT、DSA imagining show that the tumor is hypervascular. The rabbit model bearing VX2 liver tumor is more suitable for endovascular interventional experimental study.
PartⅡThe preparation of 188Re labelled Lipiodol, MAA, stannous sulfur colloid
Objective To study the methods for labeling lipiodol, macroaggregated albumin(MAA), and stannous sulfur colloid with rhenium-188.
Methods①Lipiodol was labeled with 188Re directly at the temperatures from 75℃to 120℃with different reaction conditions. The time-variation labeling efficiency was measured to evaluate the stability of 188Re-lipiodol.②Macroaggregated albumin(MAA)was labeled with 188Re after extra adding stannous chloride into commercial MAA kit, The time-variation labeling efficiency was measured to evaluate the stability of 188Re-MAA.③Different amount of stannous chloride and sodium thioslphate were respectively added into commercial sulfur colloid (SC). Their labeling efficiencies and the in vitro stabilities were tested.
Results①The optimal condition was obtained with the labeling efficiency over 99.11±0.22% at 120℃. More than 90.9±1.9% and 92.8±1.2% of radioactivity was kept in 188Re-lipidol form in room temperature and in 37℃human plasma for 72 hours;②At the condition of additional dosage of 0.5ml stannous chloride(20mg/ml), the labeling efficiency of 188Re-MAA reached(99.64±0.09)%, and(92.53±1.20)% of labeling efficiency was kept in human plasma for 72 hours③At the condition of additional dosage of 0.5ml stannous chloride(20mg/ml) and 0.25ml sodium thioslphate (8mg/ml), the labeling efficiency of 188Re-SC(Sn) reached(98.39±2.66)%, and labeling efficiency was (93.62±0.64)% in the solution (suspending SC to saline=1:10) in room temperature at 72h after preparation.
Conclusion The methods for labeling lipiodol, macroaggregated albumin(MAA), and stannous sulfur colloid with rhenium-188 are convenient,and stable in vitro with higher labeling efficiency, which can potentially be the new radiopharmaceuticals used for the treatment of stereo tumor.
PartⅢThe Biodistribution after Endovascular Injection of Rhenium-188 Labeled Drugs in Rabbit VX2 Liver Tumor Model
Objective To investigate the biodistribution property of rhenium-188 labeled drug after transcatheter arterial injection of rhenium-188 labeled lipiodol, rhenium-188 labeled macroaggregated albumin(MAA), and rhenium-188 labeled stannous sulfur colloid in rabbit VX2 liver tumor model.
Methods 64 rabbits bearing liver tumor were performed transcatheter hepatic arterial injection of lipiodol (group A), Na188ReO4 solution (group B), rhenium-188 labeled lipiodol (group C), rhenium-188 labeled MAA (group D),and rhenium-188 labeled stannous sulfur colloid(group E), respectively. The rabbits in group B、C、D and E were accepted SPECT, then sacrificed at the time of 1h or 24h after administration . Samples of brain, heart, lung, liver, spleen, kidneys, intestine, muscle, blood, and tumor were taken, weighed and measured for radioactivity. The radioactivity of tissue was expressed as percentage of the injected dose per gram of wet tissue weigh(%ID/g). The ratios of tumor to liver, kidneys, muscle, blood, lung were calculated. The ALT、AST and UREA of each of rabbit were detected before and at the time of 1h、24h after administration, respectively.
Results 35 rabbits were accepted endovascular administration, and accomplished in the study. The SPECT imaging of 23 rabbits (group C, n=6; group D, n=9; group E, n=8) shew that radioactivity was accumulated in tumor at the time of 1h or 24h after administration. The biodistribution of rhenium-188 labeled drug shew that higher radioactivities uptake were in tumor, liver and kidneys, lower radioactivities uptake were in intestine, heart, brain and muscle. and radioactivities uptake were moderate in lung, blood, and spleen. Radioactivities were decreased as the time passed. The uptake of radioactivity in tumor was higher than in the other tissues. The ratios of T/NT (tumor/ liver, tumor/ kidney, tumor/ lung, tumor/ blood, and tumor/muscle) in each group were compared. There were statistics differences among group B、C、D and E(p<0.05)each other. Except for group C, the differences of the ratio of tumor/ liver were significant between the time of 1h and 24h after administration in group D and E(p<0.05). In each group, the levels of ALT and AST even rose to the highest at the time of 24h after administration, and the amplitude in group B was the smallest. There were no statistics differences among group A、C、D and E(p>0.05), and the differences were significant between group B and group A、C、D、E(p<0.05), respectively. There were statistics differences between the both of the different time, too. In each group, the levels of UREA weren’t obviously changed.
Conclusion After transcatheter arterial administration, rhenium-188 labeled lipiodol, rhenium-188 labeled MAA, rhenium-188 labeled stannous sulfur colloid has better tumour affinity and retainable accumulation characteristics in tumor which can potentially be the new radioactive agents used for endovascular treatment of liver cancer.
引文
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