脂膜微囊承载紫杉醇靶向性治疗C6胶质瘤的体内外实验研究

The targeted therapeutic effect of a lipid-coated microbubble carrying Taxol on C6 glioma in vivo and vitro

目的:在体内外研究脂膜微囊(LCM)承载紫杉醇(Taxol)治疗大鼠颅内C6胶质瘤的机制。方法:体外实验应用紫杉醇-脂膜微囊(Taxol-LCM)或单用LCM处理C6细胞系,采用免疫荧光显微镜或激光共聚焦显微镜下观察LCM在肿瘤细胞内的分布特点,用药前后肿瘤细胞形态的改变和细胞内超微结构的变化,动态观察LCM进入细胞的全过程;体内实验建立大鼠C6胶质瘤动物模型12只,成瘤大鼠分为4组:注射LCM-Taxol组、注射LCM组、注射Taxol组及载瘤动物组,每组取一只在最后一次尾静脉注射相应药物后处死,获取大鼠脑组织标本,进行油红O染色,观察LCM在肿瘤位点分布的特点;同时观察不同处理组大鼠的生存期。结果:体外实验表明,Taxol可结合于LCM并被胶质瘤细胞内吞,起到很强的的杀伤肿瘤细胞作用。其内吞过程为LCM先结合于细胞膜表面,后在细胞内重新分布,最终被细胞浆内的酸性成分所降解。体内油红O染色显示,注射LCM-Taxol组肿瘤区域有LCM聚集,胶质瘤明显坏死;而注射LCM组,肿瘤区域有LCM出现,但肿瘤区域无明显坏死;注射Taxol组及载瘤动物组肿瘤区域无明显坏死;生存期结果示,Taxol-LCM组生存期明显长于其他3组,而与其他3组生存期比较差异不明显。结论:LCM可承载Taxol在体内外起到杀伤胶质瘤细胞的作用。

Objective： brain C6 glioma in vitro and vivo. LCM. The distribution of LCM and To investigate the therapeutic effect of lipid-coated microbubbles carrying Taxol on rat Methods： For the in vitro study, C6 glioma cell lines were treated with Taxol-LCM or changes of cell morphology and ultra-structure were observed by immunoflurescent microscopy and confocal microscopy. The process of LCMs entering the glioma cells was dynamically observed. For the in vivo study, 12 rats with intra-cranial C6 glioma were divided into 4 groups： groups with LCM-Taxol injection, LCM injection, Taxol injection and the control group. For the in vivo study, each rat was sacrificed to harvest the brain. Oil red 0 staining was used to determine the therapeutic effect and distributional characters in intlra-cranial C6 glioma. Life spans were also compared. Results： For the in vitro study, Taxol was connected with lipid-coated micro-bubbles and the com- pound was endocyted by C6 glioma and had a killing effect. The whole endocyte process was that lipid- coated microbubbles first connected with the surface of glioma cells, second were redistributed in the cells, and finally degraded in the cells associated with the acidic component of cytoplasm. Oil red O staining revealed that LCM had aggregated and there was obvious necrosis in the tumor region in the LCM-Taxol injection group. In the LCM injection groups, there was LCM in tumor regions without obvious necrosis. In the Taxol injection group and the control group, there was no nectrosis or LCM. Life span comparison revealed that survival of the LCM-Taxol injection group was much longer than that of the other three groups. Conclusion： Lipid-coated microbubbles may carry Taxol to C6 glioma cells, and the compound may have a killing effect in vitro and in vivo.