【摘要】：目的原發(fā)性肝癌是危害人類生命健康最常見的惡性腫瘤之一，由于發(fā)病隱匿，惡性程度高，易發(fā)生浸潤和轉(zhuǎn)移，發(fā)現(xiàn)時大多已經(jīng)是中晚期，失去手術(shù)治療的機會，目前迫切需要尋找到行之有效的治療方法。超聲介導(dǎo)的微泡靶向治療是近年來研究的熱點。本實驗通過制備載多西紫杉醇脂質(zhì)微泡，并通過體外實驗考察此微泡聯(lián)合超聲對人肝癌HepG2細(xì)胞的增殖抑制及誘導(dǎo)凋亡作用，探索靶向治療肝癌的可行性和有效性。 方法本實驗采用磷脂類化合物為膜材料，包裹氣體選擇C3F8，藥物為多西紫杉醇，采用薄膜水化法制備載藥超聲微泡。以包封率為指標(biāo)，通過單因素考察磷脂與多西紫杉醇藥物的比例，磷脂與膽固醇的比例、旋轉(zhuǎn)蒸發(fā)的溫度、水化的溫度和時間、機械震蕩的時間篩選出最佳制備工藝；掃描電鏡和透射電鏡觀察微泡形態(tài)及檢測粒徑大小，HPLC法檢測包封率和載藥量，4℃和25℃條件下考察其穩(wěn)定性，超聲觀察體內(nèi)顯影效果。體外實驗中，采用CCK-8法檢測多西紫杉醇對人肝癌HepG2細(xì)胞的生存影響；CCK-8法檢測各實驗組對細(xì)胞的增殖抑制作用；倒置顯微鏡觀察各實驗組細(xì)胞凋亡情況；流式細(xì)胞儀檢測各實驗細(xì)胞的周期及凋亡；透射電鏡觀察各實驗組細(xì)胞的微結(jié)構(gòu)變化。 結(jié)果載多西紫杉醇脂質(zhì)微泡外形圓整，分散好，無黏連，大小均勻，，粒徑范圍在200~650nm之間；包封率為(85.72±2.68)%，載藥量為(14.29±1.17)%；4℃條件下保存10d性質(zhì)穩(wěn)定，其粒徑在230~700nm之間；包封率為(80.25±2.26)%，載藥量為(13.37±1.13)%；體內(nèi)造影中超聲微泡能夠增強顯影效果，增大超聲儀MI微泡能順利被擊碎。體外實驗結(jié)果顯示：多西紫杉醇能明顯影響人肝癌HepG2細(xì)胞的生存，且細(xì)胞的存活率與藥物濃度和作用時間呈量效關(guān)系。載多西紫杉醇脂質(zhì)微泡聯(lián)合超聲對人肝癌HepG2細(xì)胞增殖抑制和凋亡誘導(dǎo)作用較其他實驗組更顯著。單純使用載多西紫杉醇脂質(zhì)微泡對人肝癌HepG2細(xì)胞沒有明顯增殖抑制和誘導(dǎo)凋亡作用，且此微泡只有在超聲作用下才能導(dǎo)致微泡內(nèi)藥物的釋放。 結(jié)論載多西紫杉醇脂質(zhì)微泡是一種安全、有效、穩(wěn)定性高的藥物載體；該微泡聯(lián)合超聲對人肝癌HepG2細(xì)胞增殖抑制和誘導(dǎo)凋亡作用比單純多西紫杉醇效果更顯著。載多西紫杉醇脂質(zhì)微泡有望成為腫瘤靶向治療的一種新型有效的給藥途徑。
[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

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