载多西紫杉醇脂质微泡制备及对人肝癌HepG2细胞抑制作用体外实验研究
发布时间：2018-11-23 20:26
【摘要】：目的原发性肝癌是危害人类生命健康最常见的恶性肿瘤之一，由于发病隐匿，恶性程度高，易发生浸润和转移，发现时大多已经是中晚期，失去手术治疗的机会，目前迫切需要寻找到行之有效的治疗方法。超声介导的微泡靶向治疗是近年来研究的热点。本实验通过制备载多西紫杉醇脂质微泡，并通过体外实验考察此微泡联合超声对人肝癌HepG2细胞的增殖抑制及诱导凋亡作用，探索靶向治疗肝癌的可行性和有效性。 方法本实验采用磷脂类化合物为膜材料，包裹气体选择C3F8，药物为多西紫杉醇，采用薄膜水化法制备载药超声微泡。以包封率为指标，通过单因素考察磷脂与多西紫杉醇药物的比例，磷脂与胆固醇的比例、旋转蒸发的温度、水化的温度和时间、机械震荡的时间筛选出最佳制备工艺；扫描电镜和透射电镜观察微泡形态及检测粒径大小，HPLC法检测包封率和载药量，4℃和25℃条件下考察其稳定性，超声观察体内显影效果。体外实验中，采用CCK-8法检测多西紫杉醇对人肝癌HepG2细胞的生存影响；CCK-8法检测各实验组对细胞的增殖抑制作用；倒置显微镜观察各实验组细胞凋亡情况；流式细胞仪检测各实验细胞的周期及凋亡；透射电镜观察各实验组细胞的微结构变化。 结果载多西紫杉醇脂质微泡外形圆整，分散好，无黏连，大小均匀，，粒径范围在200~650nm之间；包封率为(85.72±2.68)%，载药量为(14.29±1.17)%；4℃条件下保存10d性质稳定，其粒径在230~700nm之间；包封率为(80.25±2.26)%，载药量为(13.37±1.13)%；体内造影中超声微泡能够增强显影效果，增大超声仪MI微泡能顺利被击碎。体外实验结果显示：多西紫杉醇能明显影响人肝癌HepG2细胞的生存，且细胞的存活率与药物浓度和作用时间呈量效关系。载多西紫杉醇脂质微泡联合超声对人肝癌HepG2细胞增殖抑制和凋亡诱导作用较其他实验组更显著。单纯使用载多西紫杉醇脂质微泡对人肝癌HepG2细胞没有明显增殖抑制和诱导凋亡作用，且此微泡只有在超声作用下才能导致微泡内药物的释放。 结论载多西紫杉醇脂质微泡是一种安全、有效、稳定性高的药物载体；该微泡联合超声对人肝癌HepG2细胞增殖抑制和诱导凋亡作用比单纯多西紫杉醇效果更显著。载多西紫杉醇脂质微泡有望成为肿瘤靶向治疗的一种新型有效的给药途径。
[Abstract]:Objective Primary liver cancer is one of the most common malignant tumors that endanger human life and health. At present, there is an urgent need to find effective treatment methods. Ultrasound-mediated microbubble targeting therapy is a hot topic in recent years. In this study, the lipids loaded with docetaxel were prepared, and the effects of the microbubbles combined with ultrasound on the proliferation and apoptosis of human hepatocellular carcinoma (HepG2) cells were investigated in vitro to explore the feasibility and effectiveness of targeted treatment of hepatocellular carcinoma (HCC). Methods using phospholipid compounds as membrane materials, encapsulated gas selected C _ 3F _ 8 and docetaxel as drug, the microbubbles were prepared by membrane hydration. With the encapsulation efficiency as the index, the optimum preparation process was selected by single factor investigation of the ratio of phospholipid to docetaxel, the ratio of phospholipid to cholesterol, the temperature of rotation evaporation, the temperature and time of hydration, and the time of mechanical oscillation. The morphology and particle size of microbubbles were observed by scanning electron microscope (SEM) and transmission electron microscope (TEM). The encapsulation efficiency and drug loading were detected by HPLC method. The stability of microbubbles was investigated at 4 鈩
本文编号：2352599
[Abstract]:Objective Primary liver cancer is one of the most common malignant tumors that endanger human life and health. At present, there is an urgent need to find effective treatment methods. Ultrasound-mediated microbubble targeting therapy is a hot topic in recent years. In this study, the lipids loaded with docetaxel were prepared, and the effects of the microbubbles combined with ultrasound on the proliferation and apoptosis of human hepatocellular carcinoma (HepG2) cells were investigated in vitro to explore the feasibility and effectiveness of targeted treatment of hepatocellular carcinoma (HCC). Methods using phospholipid compounds as membrane materials, encapsulated gas selected C _ 3F _ 8 and docetaxel as drug, the microbubbles were prepared by membrane hydration. With the encapsulation efficiency as the index, the optimum preparation process was selected by single factor investigation of the ratio of phospholipid to docetaxel, the ratio of phospholipid to cholesterol, the temperature of rotation evaporation, the temperature and time of hydration, and the time of mechanical oscillation. The morphology and particle size of microbubbles were observed by scanning electron microscope (SEM) and transmission electron microscope (TEM). The encapsulation efficiency and drug loading were detected by HPLC method. The stability of microbubbles was investigated at 4 鈩
本文编号：2352599
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