眼镜蛇毒细胞毒素缓释微球瘤内注射对裸鼠人肝癌皮下移植瘤的影响  

Effects of intratumoral injection of microspheres containing cobra venom cvtotoxin on transplanted human hepatoma in nude mice

作　　者：王艳[1] 林礼务[1] 陈志奎[1] 薛恩生[1] 林晓东[1] 俞丽云[1] 林振湖[1] 

机构地区：[1]福建医科大学协和医院超声科福建省超声医学研究所,福建福州350001

出　　处：《中西医结合学报》2009年第9期836-841,共6页Journal of Chinese Integrative Medicine

基　　金：福建省卫生厅青年科研课题资助项目(No.2007-1-5)

摘　　要：目的:评价眼镜蛇毒细胞毒素(cytotoxin,CTX)-聚乳酸-羟基乙酸(polylactic-co-glycolic acid,PLGA)微球经皮瘤内注射治疗裸鼠肝癌的安全性和有效性。方法:采用甲基噻唑基四唑法检测分离纯化得到的眼镜蛇毒CTX体外细胞毒活性,复乳-溶剂挥发法制备眼镜蛇毒CTX-PLGA缓释做球。40只BALB/c裸小鼠皮下接种人肝癌细胞系BEL-7404细胞建立裸鼠皮下移植瘤肝癌模型,分为生理盐水组、空白微球组、游离CTX组和CTX-PLGA微球组。瘤内注射相应药物,治疗后每周用高频超声观察肿瘤内部回声和血流情况;治疗后第26天,剥离肿瘤称取质量并计算抑瘤率;取肿瘤组织和心、肝、肾等重要脏器,观察其病理学改变。结果:眼镜蛇毒CTX对体外培养的人肝癌:BEL-7404细胞具有较强的毒性作用。眼镜蛇毒CTX-PLGA缓释微球粒径约(34.45±9.85)μm,具有较高的包封率和较好的缓释效果。瘤体内一次性注射眼镜蛇毒CTX-PLGA微球后,肿瘤体积增长缓慢,抑瘤率达52.36%;光学显徽镜观察显示肿瘤组织大部分坏死,但心、肝、肾等重要脏器在形态学上无明显改变。结论:眼镜蛇毒CTX-PLGA微球瘤内注射可抑制裸鼠肝癌的生长,且具有较高的安全性。Objective： To evaluate the safety and efficacy of intratumoral injection of polylactic-co-glycolic acid （PLGA） microspheres containing cobra venom cytotoxin in nude mice with transplanted human hepatoma. Methods： Cytotoxic activity of cytotoxin from cobra venom was determined by using methyl thiazolyl tetrazolium method in vitro. Microspheres containing cobra venom cytotoxin were prepared with a double emulsion-solvent evaporation method. Forty BALB/c nude mice were inoculated subcutaneously in right flank with hepatoma BEL-7404 cells. Thirty-two mice whose tumor size reached about 1.0 cm in diameter, were randomly assigned into normal saline group, blank microsphers group, cytotoxin group and cytotoxin-PLGA group. Nude mice were intratumorally injected with normal saline, blank microspheres, cytotoxin or cytotoxin-PLGA microspheres respectively. Internal echo characteristics and blood flow of tumors were observed by high-frequency ultrasound every week after treatment. Twenty-six days after treatment, the tumors were removed to calculate the inhibition rate of tumor growth. The tumor, heart, liver and kidney tissues were obtained for histopathological examination. Results： The cytotoxin separated and purified from crude cobra venom caused intense cytotoxic effects to the BEL-7404 cells in vitro. The diameter of PLGA microspheres containing cobra venom cytotoxin was about （34.45±9.85） μm. Encapsulation rate was up to （78.13±8.92）%, and cumulative amount of cobra venom cytotoxin released from the PLGA microspheres in vitro during 30 days was up to 84.3%. After intratumoral injection, tumor volumes and weights in the cytotoxin-PLGA group were lower than those in the normal saline group, with a tumor growth inhibition rate of 52.36%. Observed under a light microscope, most tumor tissues were necrotic. No obvious morphological change could be seen on the liver, kidney and heart tissues. Conclusion： The above findings indicate that intratumoral injection of cytotoxin-PLGA microspheres has str

关 键 词：眼镜蛇毒细胞毒素 肝肿瘤 药物疗法 缓释制剂 小鼠 裸 

分 类 号：R735.7[医药卫生—肿瘤]