目的: 制备纳米级的载紫杉醇促黄体激素释放激素(luteinising-hormone releasing hormone,LHRH)靶向脂质微泡超声造影剂并评价其理化特性。方法: 通过旋转蒸发法、机械震荡法及生物素-亲和素法制备LHRH靶向脂质微泡。Malvern检测仪检测其粒径及表面电位,超声下观察其体外显影情况,高效液相色谱法检测紫杉醇的载药量及包封率,流式细胞仪检测二抗分别与靶向微泡、非靶向微泡的结合率。结果: 靶向脂质微泡外观圆整,分布均匀,在室温下保存14 d后,微泡的一般特性与制备初比较,差异无统计学意义(P>0.05)。靶向微泡及非靶向微泡与二抗的结合率分别为(97.76±2.18)%、(6.48±0.71)%,二者比较差异具有统计学意义(P<0.05)。结论: 成功制备纳米级载紫杉醇靶向脂质微泡,此微泡粒径较小,稳定性较高,且LHRH的靶连接率较高。
Objective: To prepare the targeted lipid-microbubble ultrasonographic contrastagent loaded with paclitaxel LHRH, and to evaluate its physical and chemical properties,Methods:LHRH targeted lipid-microbubble was prepared by Co-evaporating method, mechanical vibration method and biotin-avidin method. The size and surface electric potential were detected by Malvern detector and the imaging was observed by ultrasonography. The drug-loading capacity and encapsulation rate were determined by high performance liquid chromatography (HPLC). The connection rates of secondary antibody with targeted and non-targeted lipid-microbubble were detected by flow cytometry (FCM).Results: The targeted lipid- microbubble had complete appearance and even distribution. After being kept at room temperature for 14 days, there was no significant difference of general properties ( P>0.05). The binding rates of secondary antibody with targeted and non-targeted lipid-microbubble were 97.76±2.18 % and 6.48±0.71 % respectively, which had significant difference (P<0.05).Conclusion: The targeted lipid-microbubble loaded with paclitaxel is successfully prepared, with smaller size and higher stability. It has a high connection rate of LHRH.
[1] Jelovac D,Armstrong DK.Recent progress in the diagnosis and treatment of ovarian cancer[J].CA Cancer J Clin, 2011,61(3):183-203.
[2] Saad M,Garbuzenko OB,Ber E,et a1.Receptor targeted polymers,dendrimers,liposomes:which nanocarrier is the most efficient for tumor-specific treatment and imaging?[J].Control Release,2008,130(2):107-114.
[3] 郝权,田菁,王耕辛.卵巢癌靶向治疗研究进展[J].中国肿瘤临床,2005,32(22):1315-1318.
[4] Ward M, Wu J, Chiu JF.Experimental study of the effects of Optison concentration on sonoporation in vitro[J].Ultrasound Med Biol.2000,26(7):1169-1175.
[5] Bast RC,Skates S,Lokshin A,et al.Differential diagnosis of apelvic mass:Imp Roved algorithms and novel biomarkers.Int J Gynecol Cancer,2012,22Suppl 1(1):S5-S8.
[6] 陈智毅,谢明星.超声靶向破坏微泡技术的应用进展[J].中国医学影像技术,2010,26(9):1790-1792.
[7] 王冬,谭开彬,杨珂,等.纳米级超声微泡造影剂的制备及其性状的初步研究[J].临床超声医学杂志,2007,(12):709-711.
[8] Willmann JK,Paulmurugan R,Chen K,et al.US imaging of tumor angiogenesis with microbubbles targeted to vascular endothelial growth factor receptor type 2 in mice[J].Radiology,2008,246(2):508-518.
[9] Engel JB,Schally AV,Buchholz S,et al.Targeted chemotherapy of endometrial,ovarian and breast cancers with cytotoxic analogs of luteinizing hormone-releasing hormone (LHRH)[J].Arch Gynecol Obstet,2012,286(2):437-442.
[10] 贺娟,孙江川,常淑芳,等.人卵巢癌靶向超声造影剂的制备及其体外寻靶能力观察[J].中国医学影像技术,2009,25(6):929-931.
[11] Reches M,Gazit E.Biological and chemical decoration of peptide nanostructures via biotin-avidin interactions[J].J Nanosci Nanotechnol,2007,7(7):2239-2245.