淫羊藿苷元脂质体的制备及其对大鼠骨髓基质细胞增殖分化影响的研究  

Preparation of Icaritin Liposome and Its Effects on Proliferation and Differentiation of Rat Bone Marrow Stromal Cell in Vitro

作　　者：贾东升[1,2] 贾晓斌[1,2] 施峰[1] 薛璟[1] 孙娥[1] 陈玲玲[1] 张振海[1] 

机构地区：[1]江苏省中医药研究院中药新型给药系统重点实验室,南京210028 [2]江苏大学药学院,江苏镇江212013

出　　处：《中国药学杂志》2010年第5期353-358,共6页Chinese Pharmaceutical Journal

基　　金：江苏省医药高技术计划项目(BG2007614);江苏省中医药领军人才项目(2006);江苏省中医药科技项目(LZ09067)

摘　　要：目的研制淫羊藿苷元(icaritin,IT)脂质体,并对其进行理化性质考察和初步药效学评价。方法采用改进的乙醇注入法制备IT脂质体,考察其形态、粒径及分布、Zeta电位等理化性质;分别采用葡聚糖凝胶柱色谱法、微柱离心法及离心超滤法分离游离药物和脂质体,HPLC测定IT的包封率;以反相透析法考察IT脂质体的体外释放;以包封率为指标,采用正交设计优化制备工艺;采用大鼠骨髓基质细胞模型,初步评价了IT脂质体体外抗骨质疏松作用。结果优化条件下制备的IT脂质体均匀圆整,平均粒径(71.6±6.9)nm,Zeta电位为-(77.3±5.1)mV,采用微柱离心法测得包封率为(91.49±1.15)%,载药量为(9.15±0.22)%,IT脂质体在体外具有一定的缓释性,72h累积释放量大于90%;初步药效学实验表明,IT脂质体能促进骨髓基质细胞向成骨细胞分化,增强成骨细胞活性,抑制骨髓基质细胞向脂肪细胞分化。结论本法制备的IT脂质体粒径小、包封率高,体外细胞实验显示了较好的抗骨质疏松活性,制备方法简单可行,IT脂质体有望开发为抗骨质疏松临床用药剂型。OBJECTIVE To prepare icaritin（IT）liposome,and investigate its physico-chemical properties and effects on proliferation and differentiation of rat bone marrow stromal cells in vitro.METHODS IT liposome was prepared by modified ethanol injection method with encapsulation efficiency as index.L16（45）orthogonal design was used to optimize the formula.Based on the optimized condition,physico-chemical properties of IT liposome was studied,including morphology,mean diameter and Zeta electric potential with a laser particle size analyzer.After the free drug was removed from the liposome solution by Sephadex G-50 column,mini column centrifugation method and super filtration,respectively.The entrapment efficiency（EE,%）of the liposome was determined by HPLC and release degree was detected by reverse dialysis.Osteoblasts induced differentiation and fat accompany phenomenon of IT liposome was inspected by CAKP dyeing and methylthioninium chloride dyeing.RESULTS The average intensity diameter,entrapment efficiency by the mini-column centrifugation method and drug loading of the optimized liposome were（71.6±6.9）nm（n=3）,91.49% and（9.15±0.22）%,respectively.72 h cumulative release of IT liposome in 20% methanol PBS solution was over 90%.IT liposomes significantly enhanced the activity of alkaline phosphatase activity in osteoblasts,and inhibited satellite phenomenon of fat cell in marrow stroma cell in some degree.CONCLUSION It was feasible to prepare IT liposome by modifying ethanol injection method,reproducibility was good,entrapment rate was high,IT liposomes have delayed release ability,initial potency showed that IT liposomes have anti-osteoporosis activity.IT liposomes can be a candidate pharmaceutical dosage form for anti-Osteoporosis.

关 键 词：淫羊藿苷元 脂质体 制备工艺 药效学 

分 类 号：R944[医药卫生—药剂学]