【作者】 关键；

【导师】 胡道予；

【作者基本信息】 华中科技大学 ， 影像医学与核医学， 2009， 博士

【摘要】 目的：改进兔VX2肝癌模型介入治疗的实验方法，分析兔VX2肝肿瘤的影像学表现。方法：开腹种植VX2瘤块于兔肝左叶，建立VX2肝肿瘤模型兔15只。2周后行CT及彩超检查证实接种成功。应用显微外科手术临时阻断肝总动脉血流，经胃十二指肠动脉留置导管行介入治疗，再恢复肝动脉血流，术后导管埋置于皮下。用多层螺旋CT(MSCT)、彩色超声和数字剪影血管造影(DSA)对兔VX2肝肿瘤行影像学评价。结果：经胃十二指肠动脉留置导管操作成功11只，麻醉过量致死及术中大出血死亡各1只，另2只因解剖变异未做插管。MSCT平扫兔VX2肝肿瘤表现为低密度结节影：彩超上瘤灶为低回声光团，肿瘤周边及内部可见较丰富血流信号；DSA图像示VX2瘤灶于动脉期呈结节状肿瘤染色，以周边染色为主。结论：经胃十二指肠动脉留置导管对兔VX2肝癌行术中介入治疗，可实现超选择血管介入治疗，对肝脏血流动力学影响较小，便于术后观察，实验人员可完全避免放射辐射，但对实验条件和实验技术有一定要求。目的：研究经导管动脉注射羟基磷灰石纳米微粒(nHAP)对兔肝VX2肿瘤的治疗作用，并了解碘油和nHAP混合物在主要脏器的分布和影响。方法：将45只成功接种肝VX2肿瘤的模型兔随机分成3组，每组15只，用显微外科手术临时阻断肝总动脉血流，经胃十二指肠动脉插管分别给药行介入治疗，术毕结扎胃十二指肠动脉。A组为生理盐水组(对照组)，注射生理盐水0．5～1．0ml；B组为碘油组(疗效对比组)，注射超液化碘油0．5～1．0ml；C组为nHAP组，注入0．5％nHAP 0．5～1．0ml。3组实验动物分别于治疗前、治疗后7、14天行多层螺旋CT(MSCT)肝脏扫描，测量肿瘤的大小，并计算肿瘤生长率。治疗前、治疗后第1、7天测血清天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)两项肝酶指标；并记录各组术后生存天数。另取6只肝VX2肿瘤的模型兔按C组方法灌注nHAP和碘油的混合物0．5～1．0ml后，定期处死(术后1h和术后24h各处死3只)，通过病理切片HE染色和透射电镜分析了解碘油-nHAP混合物对主要脏器的影响和分布情况。结果：术后7、14天，A组动物的肿瘤生长率分别为350±116％和1098±337％、B组为234±18％和730±32％，C组为233±15％和723±30％。B、C两组与A组相比差异有显著性(P＜0．05)，B组与C组之间无明显差异(P＞0．05)。术前各组血清AST、ALT水平差异无统计学意义(P＞0．05)，术后1天，A、B、C组血清AST、ALT均升高，B、C组上升幅度稍高，但与A组相比差异无统计学意义(P＞0．05)。术后7天，各组AST、ALT均降至正常范围。A、B、C组瘤兔的生存期分别为38．0±5．4d、54．0±8．1d、56．0±8．2d，B、C两组与A相比生存期明显延长(P＜0．05)。光镜下HE染色见肝内肿瘤组织表现为癌巢和坏死结构，癌旁肝组织、心、肾、肺、脾等均无异常。透射电镜显示术后1h和24h兔肝肿瘤组织中均可见nHAP沉积，而其他脏器内均未见nHAP分布。结论：经动脉灌注nHAP治疗兔肝VX2肿瘤能有效抑制肿瘤生长，延长瘤兔的生存期，而无明显肝功能损害；经动脉灌注碘油和nHAP的混合物后，药物主要分布于肿瘤组织，而其他重要脏器内未见nHAP分布，亦未见相关并发症发生。目的：通过影像学和病理学评价经胃十二指肠动脉注射5-FU缓释微球对兔VX2肝肿瘤的治疗作用。方法：将成功接种肝VX2肿瘤的模型兔随机分成4组，每组10只，用显微外科手术临时阻断肝总动脉血流，经胃十二指肠动脉插管至肝固有动脉起始部给药行介入治疗，术毕结扎胃十二指肠动脉。A组(生理盐水对照组)，注射生理盐水0．5～1ml：B组(碘佛醇对照组)，注射碘佛醇0．5～1ml；C组为碘油组(疗效对比组)，注射超液化碘油0．5～1．0ml；D组为5-FU缓释微球组(实验组)，注射5-FU缓释微球10mg和碘佛醇1ml的混合溶液。4组实验动物于治疗1周后观察肿瘤的生长情况，坏死程度，并采用原位末端标记法(TUNEL)检测肿瘤细胞凋亡指数(AI)。结果：治疗1周后D组(5-FU缓释微球组)肿瘤生长受到抑制，肿瘤生长率低于A组(生理盐水对照组)及B组(碘佛醇对照组)(P＜0．05)，与C组(碘油组)相比无显著性差异(P＞0．05)。4组肿瘤均有不同程度的坏死，D组(5-FU缓释微球组)肿瘤坏死率明显高于A组(生理盐水对照组)和B组(碘佛醇对照组)(P＜0．05)。A组(生理盐水对照组)、B组(碘佛醇对照组)和D组(5-FU缓释微球组)肿瘤细胞凋亡指数分别为1．69±0．18、1．75+0．27和8．03±0．63，5-FU缓释微球组与两对照组相比差异有统计学意义(P＜0．05)。结论：5-FU缓释微球经动脉灌注可抑制肝肿瘤生长，诱导肿瘤细胞凋亡，促进肿瘤坏死，是一种有效的化疗栓塞剂。目的：研究经胃十二指肠动脉灌注重组改构肿瘤坏死因子(TNF)对兔VX2肝肿瘤的疗效。方法：将成功接种肝VX2肿瘤的模型兔随机分成3组，每组10只，用显微外科手术临时阻断肝总动脉血流，经胃十二指肠动脉插管至肝固有动脉起始部，行介入治疗，术毕结扎胃十二指肠动脉，重新开放肝总动脉血流。A组(空白对照组)，注射生理盐水0．5～1ml：B组为碘油组(疗效对比组)，注射超液化碘油0．5～1．0ml；C组为重组改构TNF组(实验组)，注射重组改构TNF 2．5万U和生理盐水的混合液0．5～1．0ml。3组实验动物于治疗1周后多层螺旋CT(MSCT)观察肿瘤的生长情况。以病理学方法评价肿瘤坏死程度，采用原位末端标记法(TUNEL)检测肿瘤细胞凋亡指数(AI)，并通过免疫组化的方法检测介入治疗后生理盐水组、碘油组和TNF组肿瘤组织的血管内皮生长因子(VEGF)表达程度和微血管密度(MVD)。结果：术后1周后C组(TNF组)肿瘤生长受到抑制，肿瘤生长率低于A组(生理盐水对照组)(P＜0．05)，与B组(碘油组)无显著差异(P＞0．05)。3组肿瘤均有不同程度的坏死，C组(TNF组)和B组(碘油组)肿瘤坏死率明显高于A组(P＜0．05)。A组(空白对照组)和C组(TNF组)肿瘤细胞凋亡指数分别为1．69±0．18和7．26±0．45，两者相比差异有统计学意义(P＜0．05)。相对生理盐水组，介入治疗后TNF组残余肿瘤组织中的VEGF表达和MVD均有所下降．结论：重组改构TNF经动脉灌注可抑制VX2肿瘤生长，促进肿瘤细胞凋亡和肿瘤坏死，并可抑制肿瘤血管新生，是一种有效的局部化疗手段。更多还原

【Abstract】 PartⅠImprovement of Transarterial Embolization of RabbitModels with VX2 Hepatoma and Imaging EvaluationObjective:To improve intra-arterial adminstration of rabbit models with VX2hepatoma and evaluate the manifestations of the VX2 hepatomas on mutislice spiralcomputed tomography (MSCT),colorful ultrasonography and digital subtractionangiography (DSA).Methods:Fifteen New Zealand white rabbits were inoculated with fragment of VX2hepatomas into the left lobe of liver.After 2 weeks,all the rabbits were proved withliver neoplasms by MSCT scanning and colorful ultrasonography.During the openlaparotomy,common hepatic artery was ligated temporarily before transarterialembolization (TAE),and a catheter was inserted via gastroduodenal artery monitoredunder a microscope and detained in rabbit’s abdomen.TAE was performed on therabbits.And then let common hepatic artery reperfuse.MSCT scanning,colorfulultrasonography and DSA were performed to describe the features of rabbit hepaticneoplasmas.Results:TAE was successfully performed in 11 rabbits.One rabbit died foroverdosage of anesthesia.Another rabbit died for massive bleeding during thesurgeon.Artery anatomy mutations existed in two rabbits,so catheterization wasquitted.VX2 hepatomas appeared as low density nodule-like shadow on MSCTimagings,and showed low echoes,higher blood flow signals on colorfulultrasonography.Trans-gastroduodenal artery angiography showed that VX2hepatomas had nodule-like tumor stain mainly around the tumor in arterial phase.Conclusion:Superselective embolization can be performed by detained catheter viagastroduodenal artery during the open laparotomy and has little impact on hepatichemodynamics.DSA can be easily used to follow up by the detained catheter.Furthermore,operators can completely avoid radiation,but the requirements of condition and technology are higher than usual. PartⅡMSCT Effect study and Safety evaluation of TransarterialEmbolization Therapy with Nan Hydroxyapatite Particle on Rabbitswith VX2 HepatomaObjective:To study the effects of nan hydroxyapatite particle (nHAP) on rabbits withVX2 hepatoma,and analyze distribution and impact of nHAP and lipidol mixture inmain organs.Methods:45 rabbits implanted liver VX2 tumors were randomly divided into 3groups.Each group had 15 rabbits.All the rabbits were proved with liver neoplasmasby mutislice spiral computed tomography (MSCT).During the open laparotomy,common hepatic artery was ligated temporarily,and then a catheter was inserted viagastroduodenal artery under a microscope.Administration was performed by thecatheter on the rabbits.Group A (control group) was administrated with 0.5 to 1.0 mlsaline;Group B (lipiodol group) was administrated with 0.5 to 1.0ml lipiodol;GroupC (nHAP group) was administrated with 0.5 to 1.0ml 0.5% nHAP.MSCT scan wasperformed to measure tumor volume on the 7th day and 14th day after theintervention.Serum levels of AST and ALT were measured on the 1st day and 7th dayafter the intervention thrapy.The survivals of VX2 rabbits were also descriped.Another 6 rabbits with VX2 hepatomas were administrated with 0.5 to 1.0ml mixtureof nHAP and lipidol,and put them to death at regular intervals (1 hour and 24 hoursafter intervention).Distribution and impact of lipidol and nHAP mixture in vitalorgans were analyzed by pathological section and transmission electron microscope.Results:The tumor growth rates of group B and C on the 7th day and 14th day aftertreatment were lower than those of control group (P＜0.05).The tumor growth ratesbetween group B and C on the 7th day and 14th day after treatment were similar(P＞0.05).The life span of both group B and group C were longer than that of thecontrol group (P＜0.05).The serum levels of AST and ALT in all three groups werelittle higher than that of before the intervention (P＞0.05).The serum levels of AST or ALT in all groups became normal level on the 7th day after the intervention thrapy.Pathological sections showed cancer nesst and necrosis structures in VX2 hepatoma,and no abnormal structures in other organs.Under transmission electron microscope,distribution of nHAP and lipidol was found only in tumor tissues at both 1 hour and24 hours after intervention,but could’t found in other organs.Conclusion:The interventional therapy with nHAP has signifigant inhibitory effecton rabbits with VX2 hepatoma,and can prolong the rabbits life span.There are noobvious liver toxicity by using the interventional therapy.Lipidol and nHAP mixturealways distribute in tumor tissues after transarterial intervention,and no nHAP can befound in other vital organs.There are no relative complications after nHAPintervention thrapy. PartⅢExperimental Study and Effect Evaluation ofTransarterial Chemoebolizasion with 5-FU Sustained-ReleaseMicrosphere on Rabbits with VX2 HepatomaObjective:To study the therapeutic efficacy of 5-FU sustained-release microspherechemoembolization through gastroduodenal artery on rabbits with VX2 hepatoma byradiology and pathology methods.Methods:40 rabbits implanted liver VX2 tumors were randomly divided into 4groups.Each group had 10 rabbits.All the rabbits were proved with liver neoplasmasby multislice spiral computed tomograpy (MSCT).During the open laparotomy,common hepatic artery was ligated temporarily,then a catheter was inserted viagastroduodenal artery under a microscope and administration was performed throughthe catheter.We ligated gastroduodenal artery and let common hepatic artery repefuseafter administration.Group A (blank control group) was administrated with 0.5 to1.0 ml saline;Group B (contrast agent group) was administrated with 0.5 to 1.0mlioversol injection;Group C (lipiodol group) was administrated with 0.5 to 1.0mllipiodol;Group D (5-FU sustained-release microsphere group) was administratedwith mixture of 10mg 5-FU sustained- release microsphere and lml ioversol.MSCTscan was performed to measure tumor volume on the 7th day after treatment.Thetumor growth rates,necrosis rates and the tumor apoptotic index (AI) were examinedand compared.Results:One week after treatment,in the 5-FU sustained-release microsphere group(group D) the tumor was significantly inhibited,tumor growth rate was lower than thegroup A and group B (P＜0.05),and was similar to group C (P＞0.05).The tumornecrosis rates of group D was singnificantly higher than those of group A and B(P＜0.05).The tumor AIs were 1.69±0.18 (group A),1.75+0.27 (group B),8.03+0.63 (group D),respectively (P＜0.05).AI of group D was singnificantly higher than thatof group A and B.Conclusion:5-FU sustained-release microsphere infusion through artery to liver caninhibit tumor growth,induce the apoptosis and aggravate the tumor necrosis.It is aneffective chemoembolization agent. PartⅣExperimental Study and Effect Analysis of TransarterialChemotherapy with Recombinant mutant TNF on Rabbits with VX2HepatomaObjective:To study the therapeutic efficacy of recombinant mutant human tumornecrosis factor (TNF) chemotherapy through gastroduodenal artery on rabbits withVX2 hepatoma.Methods:30 rabbits implanted liver VX2 hepatoma were randomly divided into 3groups.Each group had 10 rabbits.All the rabbits were proved with liver neoplasmasby multislice spiral computed tomograpy (MSCT).During the open laparotomy,commonhepatic artery was temporarily ligated,and then a catheter was inserted viagastroduodenal artery under a microscope and administration was performed throughthe catheter.We ligated gastroduodenal artery and let common hepatic artery repefuseafter administration.Group A (blank control group) was administrated with 0.5 to 1.0ml saline;Group B (lipiodol group) was administrated with 0.5 to 1.0ml lipiodol;Group C (recombined TNF group) was administrated with mixture of 2.5×10~4 IUrecombinant mutant TNF and 1ml saline.MSCT scan was performed to measuretumor volume on the 7th day after intervention.The tumor growth rates and necrosisrates were measured.Tumor apoptotic indexs examined by TUNEL were comparedbetween group A and C.VEGF expression and MVD of all groups were detected byimmunohistochemistry.Results:One week after treatment,in the recombinant mutant TNF group (group C)the tumor was significantly inhibited,tumor growth rate was lower than the group A(P＜0.05),and was similar to group B (P＞0.05).Tumor necrosis of group A wassignificantly different from group B and C,and the necrosis rate of group B and Cwas higher than group A (P＜0.05).Tumor apoptotic indexs (AI) were 1.69±0.18 ingroup A and 7.26±0.45 in group C,and there was singnificant difference betweengroup A and group C (P＜0.05).Vascular endothelial growth factor (VEGF) expression and microvessel density (MVD) of group C were singnificantly decreased(P＜0.05).Conclusion:Recombinant mutant human TNF chemotherapy through artery to livercan inhibit tumor growth,reduce vescular growth and induce apoptosis and/or tumornecrosis.Recombinant mutant human TNF is an effective chemotherapy agent forrabbits with VX2 hepatoma.更多还原