【作者】 杨莉；

【导师】 耿宝琴； 雍定国；

【作者基本信息】 浙江大学 ， 药理学， 2002， 硕士

【摘要】 前言：自从60年代蒽环类抗肿瘤抗生素应用以来，为化疗药物开拓了一个广阔的前景，从生物合成到结构修饰改造，先后发现柔红霉素、阿霉素、表阿霉素、吡喃阿霉素、阿克拉霉素和去甲氧柔红霉素等，这类药物应用于临床，发现对许多实体瘤及血液系统恶性肿瘤均有良好的疗效。其中阿霉素被认为是一支最为有效和广谱抗肿瘤的蒽环类抗癌药物，以其作为蒽环类抗叶瘤抗生素的代表药物对其心脏毒性研究有一定的价值。阿霉素，又称14—羟基柔红霉素，具有抗瘤谱广、治疗指数高的特性，主要应用于血液系统的恶性肿瘤和实体肿瘤的化疗中。1968年，意大利的科学家从突变的streptomyces peucetius的酵解产物中分离出阿霉素(人们最早是从S．peucetius中提取柔红霉素)，并用于乳腺癌的治疗。结果发现，阿霉素有比柔红霉素强的抗鼠肿瘤作用和较好的治疗效果。甚至到现在，阿霉素仍然是一支最广谱的抗癌药物之一。但大量动物和临床实验均证明，阿霉素在治疗的同时给患者带来了许多急性和慢性的副作用，影响了药物的疗程，降低了药物的疗效。急性的副反应有，骨髓抑制，恶心，呕吐和心律失常等，但均为可逆性的改变，且临床的处理也较为容易。慢性的副反应有主要是进展性心肌损伤和最终出现的充血性心力衰竭，该反应一般是非可逆性且临床预后很差。从过去患者的资料可知，阿霉素所引起心脏毒性的改变是与其累积总剂量有关：当阿霉素累积剂量＞500mg／m~2时，就会出现心脏的毒性。所以，若不 浙江大学医学院硕士学位论文 改善阿霉素对心脏毒性的状况，它的使用前景将不容乐观。于是，人们开始致力于各种保 护的方法，尤其是对心脏毒性保护剂的研究。氨磷汀（Amifostine）从4400多种化合物中 脱颖而出，成为一个较为理想的细胞保护剂。大量的动物和临床实验均表明，氨磷汀对放 疗和化疗都有一定的保护作用。 目的：本实验通过使用阿霉素所建立的心脏毒性动物模型基础上，观察氨磷汀 （am i fostine）使用后大鼠体内清除自由基的酶（GSH－Px和SOD）的活性的变化、大鼠心 肌病理切片和大鼠左心室功能的改变，以此来研究氨磷汀是否可以达到保护阿霉素所导致 的心脏毒性作用以及若有保护作用其可能机制。 方法 SD大鼠分 3组，对照组（隔天腹腔注射 NS 3mg／kg，共 6次，n＝6），阿霉素组（隔 天腹腔注射ADM 3ms／ks，共6次，累积剂量为 18ms／ks，n＝8），氨磷汀＋FII霉素组（前3针 方法同阿霉素组，从第 4针起 ADM给予的前 30分钟先腹腔注射氨磷汀 200mg／kg，氨磷汀 累积剂量是600mg／kg，阿霉素是18mg／ks n＝8）。最后一次给药的24h后，用酶学方法测定 SOD和GSH－PX，离体心赃主动脉逆行灌流、借以PCLAB测定心功能，以及用HE染色检测心 肌细胞及心肌纤维的病理变化。 统计处理：各组数据以均数土标准误（X士S了）表示，采用 邪％8刀软件包 OnewayANOVA做分析处理，多组间比较用单因素方差做假设检验。 结果： （1）左心室功能评价：通过离体心脏 Lansendorff e流、Medlab生物信号采集处理系统 记录左室收缩曲线，结果发现，氨磷汀＋阿霉素组可显著减弱阿霉素组对心室收缩与舒张功”i 自的抑制。左心室发展压氨磷汀＋＊霉素组（n”6，89·53士10·04 mmHs）与阿霉素组（n“6， sl．71士17．47 mmH＋）相比，最大和最小收缩／舒张速率氨磷汀＋阿霉素组（n二6，1972．30 士413．78 mmHg／min，－1498．gS士294．90 mHg／min）与阿霉素组（n二6，1126．14士467．79 。mHg／。in，－821．30士292．62 mmHg／min）相比，发展压与心率的乘积氨磷汀＋FhJ霉素组（n＝6， 15266．79士3606．85 mlnHg·times／min）与阿霉素组（n二6，9516．们士4677．15 。mHg·times／min）相比可明显提高（分别是p＜0刀5，P＜0刀1，P＜0刀1和 p＜0刀5）。但氨磷汀＋ 2 浙江大学医学院颀士学位论文 阿霉素织与对照组相比均无明显差异。 （2）谷脱甘肋过氧化物酶阿霉素组（GSH－PX）的活性：阿霉素组的心脏和肝脏组织 GSH－PX （n二6，38石9t5．24活力单位和 99．spel6．］3活力单位）均明显卜降，与对照组相比分别降 低了 79％和 77％；而阿霉素＋氨磷汀纵0脏和肝脏组织的“H－Px（n二6，143．93t59．50活力单 位和 23刀8f77．99活力单位）与对照组相比，均无明显差异，GSH－PX活性分别与对照组相 比降低了30％和引％，但与阿霉素组相比分别提高了49％和26％（p＜005）。 （）超氧化物歧化酶（SOD）的活性：心肌和红细胞提取液中 SOD活性的测定结果表明， 阿霉素组酶活性（n＝6，Zll．48128．58 NU／mgprot和23更多还原

【Abstract】 INTRODUCTION: Since 1960s’,anthracycline opens up a broad prospect for chemotherapy. From biosynthesis to structure modification, scientists had found a series of anthracyclines, such as daunorubicin, doxorubicin (also called adriamycin), epirubicin, THP, pirarubicin, adarubicin, idarubicin etc. It has already been affirmed that they are all have some effects against human malignancies.Doxorubicin (14-hydroxy-daunomycin,DOX) is considered to be one of the most potent, broad-spectrum drugs against a wide range of tumor. Dox is highlyeffective against hematological malignancies and solid tumors. In 1968, dox was originally isolated form a mutant streptomyces peucetius obtained from daunorubicin-producing organism, S.peucetius. Dox showed grearter antitumor activity than daunorubicin against some murine cancers and also had a better therapeutic index. However, its clinical usage is often limited by the undesirable acute and chronic side-effects. The acute side-effects such as myelosuppression , nausea, vomiting and arrhythmias are either reversible or clinically manageable. The chronic side effects represented by the development of cardiomyopathy and ultimately congestive heart failure, which are irreversible and have bad prognosis. In a retrospective study of patient records, these cardiotoxic effects have been found to correlate with the total amount of the drug administered. Consequently, once the cumulative dose exeeds 500mg/m2, the cancer patients often were conflicted with doxombicin-induced cardiotoxicity, so, if we couldn’t protect the heart from doxombicin, it will be excluded from many chemotherapentic regimens. Thus, people would have a good way to protect the heart, but not alter the antitumor activity of doxombicin.AIM: This study aims to determine the effects of amifostine on the doxombicin-induced rat cardiotoxicity, The activity of SOD and GSH-Px, the change rat left ventricular function and cardiomyopathy are observed to explore its possible mechanism.METHODS: SD rats weighing 250?0g were divided into three groups. Control group: NS(n=6,3mg/kg) were injected ip, twice a day, cumulative dose was 18mg/kg. Dox group: DOX(n=8, 3mg/kg). were injected ip, twice a day, cumulative dose was 18mg/kg. Amifostine adding Dox group: injecting amifostine (n=8,200mg/kg,ip) before Dox administered 30 min, twice a day, from d7 to dll,it meaned started injection of amifostine after Dox cumulative dose was given 9mg/kg. After 24h at the last injection, killed rats and detected SOD and GSH-Px, left ventricular function and observed the pathological change of cardiac tissue.RESULT: (l)Hymodynamic effects:We found no significant hemody-namic effect for amifostine adding doxorubicin. LVDP, 眃p/dtmax and LVDPxHR did not differ from control values when the rats ofamifostine+dox group were perfused. However amifostine+dox could significant enhancing hemodynamic effects(n=6, LVDP89.53 + 10.04 mmHg p<0.05, +dp/dtmax 1972.30 ?413.78mmHg/min p<0.01,-dp/dtmax -1498.98 ?294.90mmHg/min p<0.01,LVDP X HR15266.79 ?3606.85mmHg ’times/min P<0.05) as compared with dox group(n=6, 51.71 + 17.47mmHg, 1126.14 + 467.79mmHg/min,-821.30 + 292.62mmHg/min,9516.71+4677.15mmHg ?times/min respectively).(2) Effect on GSH-Px activities: The activities of GSH-Px of heart and hepatic tissue were enhanced significantly in pretreatment of amifostine (n=6, 143.93?9.50U and 213.08?7.99U,about 49% and 26% increment) compared with doxirubicin group (n=6, 38.69?.24U and 99.89 ?6.13U,p<0.05).(3) Effect on SOD activities:The activities of SOD of heart and Rbc extraction were increased significantly (p<0.05 and p<0.01 respectively) in amifostine+doxorubicin group(n=6, 349.35?137.21NU/mg ?prot and 34527.00?6398.50NU/gHb, about 59% and 35% increment) compared with dox (n=6,211.48?8.58 NU/mg ?prot and 23553.31?186.58 NU/g Hb respectively).(4) Pathological observation:The changes of pathologic was observed.Dox group was diffuse cells damage with vacuolization, mergence, frank necrosis. Amifostine+doxorubicin was a更多还原