多柔比星的毒性研究进展

多柔比星是一种具有细胞周期非特异性杀伤作用的蒽环类抗肿瘤抗生素,因其具有抗肿瘤谱广、活性强等特性,广泛用于治疗各种肿瘤。多柔比星主要用于急性白血病的治疗,对乳腺癌、肺癌、膀胱癌等多种肿瘤也有一定的疗效,但其强烈的细胞毒性作用对机体可产生广泛的生物化学反应,随着药物累积剂量的增加,其不良反应的发生率也相应增高。f     

共振瑞利散射法测定多柔比星脂质体中游离多柔比星的含量

目的:建立共振瑞利散射法(RRS)测定多柔比星(DOX)脂质体中游离 DOX 含量。方法:在 pH 2.6的 Britton-Robinson(BR)缓冲液中,刚果红(CR)与 DOX 通过分子间作用力形成离子缔合物,在λ_(ex)=λ_(em)=380 nm 波长时能使 RRS 信号显著增强。结果:RRS 法在1～12 μg·mL~(-1)范围内呈线性关系,检出限为0.05~(-1)g·mL~(-1)。对6个不同批号的 DOX 脂质体混悬液中游离 DOX 的含量测定,结果与紫外可见分光光度法相符,RSD(n=6)为1.9%～3.2%,平均回收率为94.3%。结论:此方法灵敏度较高,可以用于测定 DOX 脂质体中游离 DOX 的含量。     

自噬在多柔比星所致心脏毒性中的作用研究进展

多柔比星是一种广谱高效的抗肿瘤药物,但其心脏毒性是限制其临床应用的主要原因之一。多柔比星致心脏毒性的机制尚不完全清楚,通常认为与活性氧增加、线粒体损伤和细胞凋亡等机制相关。细胞自噬是一类依赖于溶酶体的蛋白质降解途径。近年来大量研究证实,自噬在多柔比星所致心脏毒性中具有重要作用。本文主要回顾自噬在多柔比星所致心脏毒性中的作用。     

吡柔比星与多柔比星的急性心脏毒性比较

［摘要］目的观察吡柔比星（THP）或多柔比星（ADM）联合化疗方案治疗恶性肿瘤疗效及急性心脏毒性。方法128例癌症患者分为ADM组63例,THP组65例; THP组又分为THP老年组15例,THP普通组50例。THP剂量为60 mg•（m2） 1, ADM为50 mg•（m2） 1,THP老年组将总量分为二次静脉滴注,其余均为总量一次静脉滴注;用药期间静脉滴注维生素C 4 g•d 1,老年THP组 加用参麦注射液100 mL•d 1。结果THP组（老年组+普通组）和ADM组有效率分别为55.4%和55.6%,两组比较差异无显著性（P＞0.05）。两组化疗后出现心电图异常8例,占6.2 %,其中ADM组6例,THP 2例。结论THP或ADM联合化疗方案治疗恶性肿瘤疗效相当,THP急性心脏毒性较低;对老年人,既往有高血压或心绞痛病史,化疗前心电图异常者应选择THP。     

多柔比星心脏毒性的研究

多柔比星(ADM,阿霉素)是1969年从链霉Str.peucetius var．caesius培养基中分离的抗癌药。注射时药物宜用5％葡萄糖或O．9％氯化钠注射液稀释,不宜用蒸馏水,静滴浓度2 mg·ml-1,应于8h内用完。混合后可发生配伍禁忌的药物有:氨茶碱,头孢噻吩,氢化可的松,地塞米松,氟尿嘧啶及肝素。由于ADM的心脏毒性明显,探讨有效的防护心脏的药物十分必要。     

脂质体多柔比星与多柔比星治疗弥漫大B细胞淋巴瘤的比较研究

目的：比较脂质体多柔比星（PLD）和多柔比星治疗弥漫大 B 细胞淋巴瘤（DLBCL）的疗效及心脏毒性。方法本试验采用开放式、随机、对照的方法进行研究。将50例弥漫大B细胞淋巴瘤（ DLBCL）患者随机分成A组（环磷酰胺、脂质体多柔比星、长春新碱、泼尼松治疗组）、B组（环磷酰胺、多柔比星、长春新碱、泼尼松治疗组）;A组PLD剂量为40mg／m2,其余环磷酰胺、长春新碱和泼尼松采用标准 CHOP 方案中的剂量。 B组环磷酰胺、多柔比星、长春新碱、泼尼松采用标准CHOP方案中的剂量。均建议联合应用利妥昔单抗化疗。每2周期化疗后复查评价疗效;每周期评价血液学、肝肾毒性、心脏毒性等。结果入组病例中A组完成25例、B组完成25例。6～8个疗程后A组有效率84．0％,B 组为76．0％,但差异无统计学意义（P＞0．05）。中位随访24．2（0．9～40．5）个月,A组总生存率及无进展生存率分别为80．0％及72．0％,B组总生存率及无进展生存率分别为84．0％及68．0％。差异无统计学意义（ P＞0．05）。化疗最常见的不良反应为脱发、手足综合征、血小板减少、粒细胞减少、贫血、肝功能损害、心脏毒性、胃肠道反应（恶心、呕吐与腹痛、腹泻）、乏力、口腔炎。手足综合征仅见于A组。 B组的脱发、血小板减少、Ⅳ度粒细胞减少、贫血、肝功能损害、心脏毒性、恶心呕吐、口腔炎发生率大于A组,两组发生率的差异有统计学意义（P＜0．05）。 B组乏力、腹痛腹泻的发生率高于A组,但是两组乏力、腹痛腹泻的发生率差异无统计学意义（ P＞0．05）。两组神经毒性的发生率相近,差异无统计学意义（ P＞0．05）。结论与多柔比星相比,含脂质体多柔比星的类似CHOP方案能明显减轻化疗所致的脱发、心脏毒性、肝脏损害、血液学毒性、口腔炎,两组化疗所致的疗效相当。     

维生素E对多柔比星所致生殖毒性雄性大鼠的保护作用

目的探讨维生素E对多柔比星致生殖毒性雄性大鼠的保护作用。方法通过一次性静脉注射多柔比星7.5 mg.kg-1制备多柔比星致雄性大鼠生殖毒性损伤模型。维生素E组(n=8)从造模前1 d起,灌服维生素E 100mg.d-1连续14 d。模型组(n=8)造模后,每日经腹腔注射0.9%氯化钠溶液1 mL。对照组(n=8)不造模,每日经腹腔注射0.9%氯化钠溶液1 mL。通过观察大鼠血清超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量和睾丸病理组织学变化评估多柔比星对雄性大鼠生殖毒性作用。结果与对照组比较,模型组大鼠血清SOD活性降低,MDA含量升高(P<0.05),睾丸重量和睾丸系数降低(P<0.05),精曲小管生精细胞明显减少,精母细胞和精子细胞退变,部分坏死、脱落。维生素E组大鼠血清SOD活性和MDA含量与对照组比较差异无显著性(P>0.05),睾丸重量和睾丸系数的变化和睾丸病理损伤程度轻于模型组。结论维生素E对多柔比星所致生殖毒性雄性大鼠具有保护作用,其药理作用机制与提高机体抗氧化能力、抑制脂质过氧化反应有关。     

多柔比星心脏毒性及其中药防治研究进展

多柔比星是一种高效广谱抗肿瘤药物,由于严重的细胞毒性,其临床应用受到一定影响,且目前缺乏有效的预防和治疗方法。该文探讨了多柔比星的心脏毒性及其作用机制,并从中药、中药活性成分、中药注射液、中成药和煎剂等方面就近年来国内外有关中药预防多柔比星心脏毒性的研究进展进行归纳总结,以期为临床防治多柔比星毒性提供参考。     

多柔比星斑马鱼胚胎心脏发育毒性表现

目的通过观察多柔比星的斑马鱼胚胎心脏毒性表现,为其作为心脏毒性评价模型提供可靠的检测指标。方法选择发育正常的6hpf(hours post fertilization)斑马鱼胚胎暴露于多柔比星2.16～34.48μmol·L-148h。显微镜观察72hpf斑马鱼心血管系统的形态学改变。通过DVC摄像系统测定心率;测量静脉窦-动脉球(SV-BA)间距,HE染色观察斑马鱼心肌结构。结果多柔比星2.16μmol·L-1组斑马鱼心血管系统形态无改变,但心率下降,为(166±5)min-1;SV-BA间距增加,为(237±13)μm。多柔比星4.31μmol·L-1可引起斑马鱼出现心包水肿,心脏畸形,心率减低,为(166±5)min-1;SV-BA间距增加,为(268±13)μm。随着多柔比星浓度增加,斑马鱼出现体长缩短、体节发育异常、脊柱弯曲、卵黄囊水肿、出血、心脏缩小等多种表型改变。心脏组织切片显示,多柔比星8.62μmol·L-1可引起斑马鱼心包腔变大、心脏缩小、心肌层变薄和心肌细胞减少。结论多柔比星对斑马鱼胚胎心脏毒性作用的表现与对哺乳动物的毒性作用表现相同,有望成为评价药物心脏发育毒性的模型。     

盐酸多柔比星制剂研究进展

目的：介绍盐酸多柔比星（Dox）各种剂型的研究进展.方法：根据国内外文献,综述了目前Dox的微球、脂质体、纳米粒、胶束、埋植小棒、凝胶等各种剂型的设计原理、制备方法、体内外研究概况以及部分制剂的临床研究结果和上市产品.结果：由于Dox的不良反应限制了其临床使用,因此其载体制剂得到了广泛研究.这些制剂大部分改变了Dox的生物分布,提高了其在局部肿瘤组织中的含量,其中脂质体制剂已经成功应用于临床,显示了一定的优越性.结论：药物载体在体内的代谢的进一步研究,提高了Dox在体内循环过程中的稳定性及对肿瘤细胞的靶向性.     

灯盏花素促进阿霉素诱导的K562细胞凋亡

目的观察灯盏花素对淋巴细胞增殖和阿霉素致肿瘤细胞死亡的影响。方法MTT法检测细胞增殖,Western blot检测p53/bcl-2表达,Histone/DNA ELISA和流式细胞仪检测细胞凋亡,ELISA检测细胞色素C释放,分光光度法检测caspase-8和caspase-3激活。结果灯盏花素能增强小鼠淋巴细胞对阿霉素的抵抗,但促进阿霉素引起的K562细胞生长抑制、细胞色素C释放、caspase-8和caspase-3激活,上调其p53/bcl-2表达比率,增加细胞凋亡。结论灯盏花素有增强阿霉素抗肿瘤作用,激活肿瘤细胞凋亡通路可能是其化疗增敏的主要机制。     

葛根提取物诱导K562细胞凋亡

目的 探讨葛根提取物(PL)对慢性粒细胞自血病(CML)细胞株K562的凋亡诱导作用及其可能的分子机制.方法 以不同浓度(0、2.5、5、10和20 mg/ml)PL处理K562细胞,24 h后光镜下观察形态学改变,MTT法测定细胞增殖抑制,Hoechest 33258荧光染色及FITC-Annexin V/PI双染法检测凋亡率变化,半定量RT-PCR及Western blot方法检测Bcr/abl、Bcl-2、p53、Fas/FasL蛋白表达.结果 PL对K562细胞有明显的增殖抑制作用,并观察到典型的细胞凋亡形态变化,细胞凋亡率与浓度呈正相关;不同浓度PL干预后Bcr/abl基因在mRNA和蛋白水平呈浓度依赖性下调,而 Bcl-2基因则无明显变化;p53表达呈浓度依赖性上调;Fas/FasL表达无明显变化.结论 不同浓度PL能有效诱导K562细胞凋亡;其机制可能通过在mRNA和蛋白水平下调Bcr/abl基因,上调p53基因表达而实现.     

阿霉素诱导的人白血病细胞系K562/A02多药抗药性(英文)

目的:研究白血病细胞多药耐药发生的机制. 方法:用逐步增加培养基中阿霉素(Dox)浓度的方法,诱导出一株耐Dox的人白血病细胞系K562/A02.用[~3H]TdR参入法测定IC_(50),HPLC法测定细胞内药物浓度,免疫组织化学法检测P-糖蛋白的表达,RT-PCR法测定mdr1基因表达,点杂交测定基因组中mdr1DNA和拓扑异构酶Ⅱ(TopⅡ)基因表达,CDNB法测定谷胱甘肽-S-转移酶(GST)活性. 结果:K562/A02对Dox、HHT、Dau、VCR、m-AMSA、VP-16具有较强的交叉耐药性,而对Ara-C耐药较弱,对5-FU、Cis和MTX不交叉耐药,表现出典型的多药耐药表型.K562/A02细胞内药物浓度明显低于K562细胞,P-170阳性表达.K562/A02细胞中MDR1基因拷贝数与K562的无差异,但mdr1mRNA高表达.TopⅡ的mRNA水平低于K562,GST的活性增高. 结论:白血病细胞多药耐药的机制与mdr1基因表达密切相关,也与TopⅡ和GST有关.     

一叶秋碱诱导K562细胞凋亡

目的研究一叶秋碱（ＳＥＣ）能否诱导Ｋ５６２细胞凋亡。方法细胞增殖抑制采用ＭＴＴ法;形态学研究采用电子和荧光显微镜;流式细胞仪和琼脂糖凝胶电泳检测ＤＮＡ断裂。结果ＳＥＣ１０～１６０ｍｇ·Ｌ－１呈剂量依赖性抑制Ｋ５６２细胞增殖（ｒ＝０９６１３,Ｐ＜００５）;ＳＥＣ作用４８ｈ后,电镜下可见细胞膜完整,核染色质边聚和凋亡小体;荧光显微镜下核染色质凝聚成点状结构;流式细胞仪出现凋亡峰;ＤＮＡ琼脂糖凝胶电泳可见“梯状”图谱。结论ＳＥＣ可诱导Ｋ５６２细胞调亡。     

Liposome-based intracellular kinetics of doxorubicin in K562/DOX cells

Liposomes can improve the intracellular concentration of cytotoxic drugs, and are regarded as a possible pharmacological approach to overcome drug resistance. The kinetic analysis of subcellular drug uptake and efflux helps to elucidate the resistance mechanism which is associated with the ATP-dependent membrane transporter P-glycoprotein (P-gp). However, there are only few reports about the intracellular kinetics of liposomes. In this work, the kinetics of drug uptake and active efflux of doxorubicin (DOX) encapsulated in liposomes in both intact cells and nuclei were studied using P-gp expressing K562/DOX cells. The results show that liposomes enhanced drug accumulation in intact cells and nuclei, and improved DOX retention in nuclei after withdrawal. Furthermore, the nuclei levels of liposomal drug rose slowly and reached a plateau after 2 h incubation, whereas the free drug reached the plateau in 15 min, suggesting that it takes time for the liposomes to get from the cytoplasm to the nuclei. Our results demonstrated that liposomes not only increase DOX levels allocated to nuclei but also extended retention in the nuclei of resistant cells.     

Circumvention of acquired resistance to doxorubicin in K562 human leukemia cells by oxatomide

We studied the effect of oxatomide, an antiallergic drug, on the resistance of K562 cells to doxorubicin. Oxatomide synergistically potentiated the cytotoxicity of doxorubicin in doxorubicin-resistant K562 cells (K562/DXR) at a concentration of 1-10 microM, but had hardly any synergistic effect on the parental cell line (K562) at the same concentration. Oxatomide inhibit P-glycoprotein pump-efflux activity and the binding of [3H]-azidopine to the cell-surface protein P-glycoprotein, in a dose-related manner. These results indicate that oxatomide reverses the multidrug-resistance phenotype through direct interaction with P-glycoprotein.     

Reversal of acquired resistance to doxorubicin in K562 human leukemia cells by astemizole

This study demonstrates that astemizole, a non-sedating anti-histaminergic drug with low toxicity in vivo, greatly potentiates the growth-inhibitory activity of doxorubicin in doxorubicin-resistant human leukemia cells (K562/DXR). Astemizole synergistically potentiated the cytotoxicity of doxorubicin for K562/DXR cells at a concentration of 0.1-3 microM in a dose-dependent manner, whereas they showed hardly any synthergistic effect in the parental cell line (K562) at the same concentration. Since doxorubicin resistance in these cells is associated with the expression of high levels of P-glycoprotein, we evaluated the effect of astemizole on P-glycoprotein activity in cytofluorographic efflux experiments with doxorubicin. Our results indicate that astemizole inhibits the P-glycoprotein pump-efflux activity in a dose-related manner. Moreover, it also inhibits the photolabeling of P-glycoprotein by [3H]azidopine in a dose-dependent manner. These findings provide a biological basis for the potential therapeutic application of astemizole as an anticancer drug either alone or in combination with doxorubicin to multidrug-resistant leukemic cells.     

卡立泊来德对K562细胞诱导的血管生成的影响

目的研究卡立泊来德(cariporide)处理对K562细胞诱导的血管生成能力的影响。方法应用MTT检测K562细胞上清液对脐静脉内皮细胞增殖能力的影响;transwell检测K562细胞上清液对脐静脉内皮细胞迁移能力的影响;基质胶血管形成法检测K562细胞上清液对脐静脉内皮细胞体外血管形成能力的影响;激光扫描共聚焦显微镜测定K562细胞的细胞内的pH;酶联免疫吸附实验(ELISA)检测K562细胞上清中血管内皮生长因子的表达水平。结果cariporide处理可以明显降低K562细胞上清液对脐静脉内皮细胞增殖,迁移和体外成管能力的诱导;cariporide处理后K562细胞的细胞内pH明显下降,分泌VEGF能力也受到抑制。结论 cariporide能抑制K562细胞的血管生成诱导能力,这种抑制是通过细胞内pH下降以及VEGF分泌减少引起的。     

Antitumor effect of arsenic trioxide in human K562 and K562/ADM cells by autophagy

Arsenic trioxide (As(2)O(3)) has been reported to have potent antitumor effects in vitro and in vivo by inducing cell death via cell cycle arrest and apoptosis in leukemia cells, but the mechanisms of As(2)O(3)-mediated cell death are not fully understood. In this study, we provided in vitro evidence that As(2)O(3) was a potent inducer of autophagy in leukemia K562 and its drug-resistant line K562/ADM cells. As(2)O(3) significantly activated autophagic cell death (programmed cell death type II) in leukemia cell lines. Numerous large cytoplasmic inclusions, abundant autophagic vacuoles, phagocytizing cytoplasm and organelles were observed in As(2)O(3)-treated cells using electron microscope. MDC-labeled autophagic vacuoles were observed by fluorescent inverted phase contrast microscopy and the enhanced MDC fluorescent staining was detected by flow cytometry in As(2)O(3)-treated cells. Furthermore, real-time quantitative RT-PCR revealed that the expression levels of Beclin-1 and LC3 genes, which play key roles in autophagy, increased in As(2)O(3) treated samples than in controls, indicating that autophagy can potentially be involved in the antitumor properties of As(2)O(3). The expression level of Bcl-2 gene, an anti-apoptotic molecule, decreased in As(2)O(3) treated samples than in controls, suggesting that Bcl-2 may be involved in accumulating Beclin-1 and triggering autophagic cell death in As(2)O(3)-treated leukemia cells. Western blotting also showed that As(2)O(3) up-regulated Beclin-1. Altogether, our data provide direct evidence that autophagic cell death is critical for the effects of As(2)O(3) on acute myelogenous leukemia cells.     

Antitumor effect of arsenic trioxide in human K562 and K562/ADM cells by autophagy

Arsenic trioxide (As₂O₃) has been reported to have potent antitumor effects in vitro and in vivo by inducing cell death via cell cycle arrest and apoptosis in leukemia cells, but the mechanisms of As₂O₃-mediated cell death are not fully understood. In this study, we provided in vitro evidence that As₂O₃ was a potent inducer of autophagy in leukemia K562 and its drug-resistant line K562/ADM cells. As₂O₃ significantly activated autophagic cell death (programmed cell death type II) in leukemia cell lines. Numerous large cytoplasmic inclusions, abundant autophagic vacuoles, phagocytizing cytoplasm and organelles were observed in As₂O₃-treated cells using electron microscope. MDC-labeled autophagic vacuoles were observed by fluorescent inverted phase contrast microscopy and the enhanced MDC fluorescent staining was detected by flow cytometry in As₂O₃-treated cells. Furthermore, real-time quantitative RT-PCR revealed that the expression levels of Beclin-1 and LC3 genes, which play key roles in autophagy, increased in As₂O₃ treated samples than in controls, indicating that autophagy can potentially be involved in the antitumor properties of As₂O₃. The expression level of Bcl-2 gene, an anti-apoptotic molecule, decreased in As₂O₃ treated samples than in controls, suggesting that Bcl-2 may be involved in accumulating Beclin-1 and triggering autophagic cell death in As₂O₃-treated leukemia cells. Western blotting also showed that As₂O₃ up-regulated Beclin-1. Altogether, our data provide direct evidence that autophagic cell death is critical for the effects of As₂O₃ on acute myelogenous leukemia cells.