透明质酸修饰的绿原酸脂质体的制备及细胞学研究

摘要

Objective To prepare hyaluronic acid(HA) targeted chlorogenic acid(HA-CA) liposome and to investigate its inhibition effect on HA receptor(CD44) high expressing A549 cells and HA receptor(CD44) low expression HepG2 cells proliferation.Methods HA-DOPE was synthesized;HA-CA liposome was prepared by thin membrane disperse method and the particle size was measured by using the dynamic light scattering particle size analyzer;the HPLC method was adopted to establish the CA in vitro contents measurement method and detect the HA-CA liposome entrapment efficiency;MTT assay was applied to detect the proliferation inhibiting effect of free CA,CA liposome and HA-CA liposome on A549 cells and HepG2 cells;the fluorescence cell uptake assay was adopted to verify the targeting effect of HA liposome.Results The average particle size of HA-CA was 219.20 nm and PDI was 0.16;the entrapment efficiency of HA-CA liposome was(85.36 ± 1.01)％;the proliferation inhibition effect of HA-CA liposome on A549 cells was significantly greater than that of CA liposome,moreover CA liposomewas greater than free CA,the proliferation inhibition effect of CA liposome and HA-CA liposome on HepG2 cells was basically similar,which was greater than that of free CA;the uptake of A549 cells on HA liposome carrying 6-coumarin(HA-C liposome) was higher than that of HepG2.Conclusion HA-CA liposome can specifically combined with the high expression HA receptor cells to achieve the active targeting effect of tumor cells.%目的 制备透明质酸(HA)靶向绿原酸(CA)脂质体,探讨其对HA受体(CD44)高表达A549细胞及低表达HepG2细胞的增殖抑制作用.方法 合成HA-二油酰磷脂酰乙醇胺(DOPE);通过薄膜分散法制备HA-CA脂质体并用动态光散射粒径分析仪测定粒径;采用高效液相色谱法(HPLC)建立CA体外水平测定方法并测定HA-CA脂质体的包封率;采用噻唑蓝(MTT)法检测游离CA、CA脂质体及HA-CA脂质体对A549细胞和HepG2细胞的增殖抑制作用;采用荧光细胞摄取实验验证HA脂质体的靶向性.结果 HA-CA的平均粒径为219.20 nm,多分散系数(PDI) =0.16;包封率为(85.36±1.01)％;HA-CA脂质体对A549细胞的增殖抑制作用明显大于CA脂质体,且CA脂质体大于游离CA;CA脂质体和HA-CA脂质体对HepG2细胞的增殖抑制作用则基本相当,均大于游离CA;A549细胞对载6-香豆素HA脂质体(HA-C脂质体)的摄取高于HepG2细胞.结论 HA-CA脂质体能与高表达的HA受体细胞特异性结合,达到肿瘤细胞主动靶向效果.