摘要
用薄膜超声法制备T-2毒素脂质体(LP-T2),用高效液相色谱(HPLC)法测定包封率,并考察测定方法的线性、加样回收率、精密度和重复性;用单因素法考察有机溶剂、磷脂种类、m(药)∶m(脂)、水合体积、m(磷脂)∶m(胆固醇)对包封率的影响及响应面法优化处方;用高压均质法对脂质体整粒,并测定粒径分布;用噻唑蓝(MTT)法检测T-2毒素和LPT2对5种肿瘤细胞和正常细胞的生长抑制作用.结果表明:LP-T2最佳制备工艺为以蛋黄卵磷脂(EPC)为膜材,薄膜超声法制备,m(药)∶m(脂)=3.5,m(磷脂)∶m(胆固醇)=3.74,4-羟乙基哌嗪乙磺酸(Hepes,20mmol/L,pH=7.4)水合,水合体积11.8mL,超声10min;高压均质整粒后平均粒径为267nm;LP-T2可显著抑制肿瘤细胞的增殖,活性与丝裂霉素相当,对正常细胞的IC_(50)值(1 881.54ng/mL)高于T-2毒素(1 427.83ng/mL),即毒副作用减小;LP-T2的包封率和粒径均符合药剂标准,具有体外抗肿瘤活性.
T-2 toxin liposome(LP-T2)was prepared by thin film ultrasonic method.Entrapment efficiency was determined by high performance liquid chromatography(HPLC)method,and the linearity,additive recovery,precision and repeatability of the method were investigated.Effects of organic solvent,phospholipid species,m(drug)∶ m(lipid),hydration volume,m(phospholipids)∶m(cholesterol)on encapsulation efficiency were investigated by single factor method,and the prescription of T-2 toxin liposome was optimized by response surface method.The liposome was granulated by high pressure homogenization and particle size distribution was determined.The inhibitory effects of T-2 toxin and LP-T2 on growth of 5 tumor cells and one normal cell were detected by thiazolyl blue(MTT)method.The results show that the best preparation process of LP-T2 are egg phosphatidylcholine(EPC)as membrance material,thin film ultrasonic method,m(drug)∶m(lipid)=3.5,m(phospholipid)∶m(cholesterol)=3.74,4-hydroxyethyl piperazine ethylsulfonic acid(Hepes,20 mmol/L,pH=7.4)hydration,hydration volume 11.8 mL,ultrasound for 10 min.The average particle size after high pressure homogenization is 267 nm.LP-T2 can significantly inhibit the proliferation of tumor cells,and its activity is similar to that of mitomycin.IC50 value of normal cell(1881.54 ng/mL)is higher than that of T-2 toxin(1427.83 ng/mL).The entrapment efficiency and particle size of LP-T2 conform to the standard of drug,and it has anti-tumor activity in vitro.
作者
董媛
张馨月
李玉杰
李雅雅
王虹霞
潘娅楠
梁兰馨
王会岩
DONG Yuan;ZHANG Xinyue;LI Yujie;LI Yaya;WANG Hongxia;PAN Yanan;LIANG Lanxin;WANG Huiyan(School of Medical Laboratory,Jilin Medical University,Jilin 132013,Jilin Province,China)
出处
《吉林大学学报（理学版）》
CAS
北大核心
2019年第1期171-176,共6页
Journal of Jilin University:Science Edition
基金
吉林省教育厅科学技术研究项目(批准号:JJKH20170420KJ)
吉林省大学生创新创业训练计划(批准号:201613706025)
吉林省大学生创新训练项目(批准号:201731)
关键词
T-2毒素
脂质体
薄膜超声法
响应面法
抗肿瘤活性
T-2 toxin
liposome
thin film ultrasonic method
response surface method
anti-tumor activity
