喜樹鹼及其奈米粒子製劑對人類肝癌細胞毒殺作用之研究

Cytotoxicity Evaluation of Camptothecin and its Solid Lipid Nanoparticles in Human Hepatoma Cell

施家雯(Jia-Wen Shih)

指導教授 ： 蔡東榮

高雄醫學大學/藥學院/藥學研究所/碩士(2004年)

摘要

喜樹鹼（Camptothecin）為一有效抗癌藥物。以固體脂質為材料利用熱均質的方式製備含喜樹鹼之固體脂質奈米粒子（Solid lipid nanoparticles, SLN）。並應用25複因子實驗設計法，探討製備這種水難溶性藥品之奈米質粒影響其粒子大小與分佈等物性的因素，以及含喜樹鹼之奈米粒子之物化性質和藥品溶離釋出情形。並將製成的含喜樹鹼之奈米粒子與溶液劑進一步研究其對人類肝癌細胞（Hep3B 和 HepG2）及DNA合成的影響。
從25複因子實驗設計法的統計結果得知Lipid type及Surfactant type為主要影響粒徑大小的變因；所製成含Camptothecin的奈米粒子其平均粒徑約在79-119 nm之間；藥品在不同處方所製成的奈米粒子其包埋率在5-26 ％；若以掃描式熱差分析儀（DSC）檢測的結果發現，藥物是以非晶型的狀態存在奈米粒子中，在體外溶離釋放方面，製備成奈米粒子的處方其溶離釋出較慢，而有緩釋的效果。
將製成之Camptothecin奈米粒子投於人類肝癌細胞（Hep3B 及 HepG2）測定其細胞毒性，其結果顯示Camptothecin所製備的奈米粒子對於人類肝癌細胞（Hep3B 及 HepG2）比Camptothecin 溶液更具細胞毒殺作用，若添加P-glycoprotein抑制劑Cyclosporin A，則對人類肝癌細胞的毒性會顯著地增加。為了進一步了解其作用機轉，以流體細胞分析儀所測得DNA含量變化的結果可得知Camptothecin所製備之固體脂質奈米粒子對於肝癌細胞（Hep3B）的作用是促使細胞週期由G2/M期直接進入Sub G1，並可能經由細胞程序化死亡（Apoptosis）的機轉而造成細胞死亡。

關鍵字

喜樹鹼 ； 奈米粒子 ； 細胞毒性

並列摘要

In this study, we used Camptothecin (CA) as a model drug for preparation in hot homogenization. camptothecin is a natural, water-insolube alkaloid. 25 factorial designs is a useful tool in order to evaluate the effect of formulation variable on physicochemical properties of solid lipid nanoparticles (SLN). The drug incorporation to SLN were to studied for their physical properties and drug dissolution., To investigate the cytotoxic mechanism and DNA synthesis of various camptothecin formulation in human hepatoma cells（Hep3B and HepG2）.
From 25 factorial designs can be found that the main effects of two factors are lipid type and surfactant type on particle size . The CA-SLN had an average diameter of about average 79-119nm.The physical physicochemical state of CA-SLN were examined by DSC, the results indicate camptothecin might exist in an amorphous state in SLN. In vitro results showed that release rate of drug from SLN would be slower than camptothecin solution. SLN might be controlled release delivery system for camptothecin. The efficiency of encapsulation of camptothecin in various nanoparticles was 5-26％.
The cytotoxicity of camptothecin solution, camptothecin added with cyclosporin A solution, CA-SLN were determined by MTS assay and flow cytometry. The results indicate that the cytotoxicity of CA-SLN was great on human hepatoma cells（Hep3B and HepG2）.To define the cytotoxic mechanism of camptothecin add with cyclosporin A on human hepatoma cells. The sub-G1 feature analyzed by flow cytometry appeared in 48 hour. Camptothecin solution and CA-SLN treatment might drove the cells at G2/M phase to apoptosis.

並列關鍵字

solid lipid nanoparticles ； cytotoxicity ； camptothecin

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