和厚朴酚脂质体的制备及其体外抗肿瘤活性的研究
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摘要
目的制备(2,3-二油酰基-丙基)-三甲胺(1,2-dioleoyl-3-trimethylammonium-propane,DOTAP)/二油酰磷脂酰乙醇胺(dioleoyl phosphoethanolamine, DOPE)包裹和厚朴酚脂质体,并对其理化性质和人肝癌高转移细胞(highlymetastatic human hepatocellular carcinoma cell, HCCLM3)的药效学进行研究。方法利用薄膜分散法制得和厚朴酚脂质体,对不同药脂比的包封率进行考察;通过纳米粒度仪检测和厚朴酚脂质体的粒径电势分布、时间稳定性;通过生物透射电镜(transmission electron microscopy, TEM)对其大小和形态学进行观测;用细胞膜红色荧光探针标记和厚朴酚脂质体,探究HCCLM3细胞对脂质体摄取的时间依赖性;用WST-1和BD凋亡试剂盒检测和厚朴酚脂质体对HCCLM3细胞的药效作用。结果和厚朴酚脂质体粒径分布均匀,平均粒径为(112.30±0.78)nm,多分散系数为(0.24±0.00),电势为(53.23±1.16)m V。药脂比为7%时包封率最高,高达(99.47±0.20)%,并且在4℃环境下可以长期稳定存在。和厚朴酚脂质体的促凋亡水平明显强于原药,差异有统计学意义(P<0.05)。结论和厚朴酚脂质体制备工艺简单易行,具有稳定的理化性质和较高的包封率,同时优于和厚朴酚原药的药效作用。
Objective To prepare the DOTAP/DOPE honokiol liposomes(liposome-HNK) and to explore their physicochemical property and pharmacodynamics to treat human hepatocellular carcinoma cells lines(HCCLM3). Methods Liposome-HNK with different lipid-drug ratio was prepared by thin film dispersed method(TFDM) and determination of encapsulation efficiency(EE) was made; the particle size distribution of liposome-HNK and the time stability were determined by Zetasizer nano ZS and transmission electron microscope(TEM), respectively; Di L fluorescence probe was used to mark the liposome-HNK and to explore the cellular uptake in a time dependent fashion; the pharmacological effects of liposome-HNK on HCCLM3 cells were detected by WST-1 and BD apoptosis kits. Results Liposome-HNK was(112.30±0.78) nm in average particle size with even size distribution, the polydispersity index(PDI) value was(0.24 ±0.00) and zeta potential was(53.23 ±1.16)m V; when the lipid-drug ratio was 7%, the encapsulation rate reached the highest, as high as(99.47±0.20)% and had a long-term stability in 4℃ environment; the apoptosis rate of liposome-HNK was significantly higher than that in free-drug group(P<0.05). Conclusions The preparation process of liposome-HNK is simple and feasible, and it has stable physicochemical property, high encapsulation rate and higher pharmaceutical effect.
Objective To prepare the DOTAP/DOPE honokiol liposomes(liposome-HNK) and to explore their physicochemical property and pharmacodynamics to treat human hepatocellular carcinoma cells lines(HCCLM3). Methods Liposome-HNK with different lipid-drug ratio was prepared by thin film dispersed method(TFDM) and determination of encapsulation efficiency(EE) was made; the particle size distribution of liposome-HNK and the time stability were determined by Zetasizer nano ZS and transmission electron microscope(TEM), respectively; Di L fluorescence probe was used to mark the liposome-HNK and to explore the cellular uptake in a time dependent fashion; the pharmacological effects of liposome-HNK on HCCLM3 cells were detected by WST-1 and BD apoptosis kits. Results Liposome-HNK was(112.30±0.78) nm in average particle size with even size distribution, the polydispersity index(PDI) value was(0.24 ±0.00) and zeta potential was(53.23 ±1.16)m V; when the lipid-drug ratio was 7%, the encapsulation rate reached the highest, as high as(99.47±0.20)% and had a long-term stability in 4℃ environment; the apoptosis rate of liposome-HNK was significantly higher than that in free-drug group(P<0.05). Conclusions The preparation process of liposome-HNK is simple and feasible, and it has stable physicochemical property, high encapsulation rate and higher pharmaceutical effect.
引文
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[6]王冠杰,赵娅娟,董济民.脂质体和厚朴酚联合热疗对腺样囊性癌增殖的协同抑制作用[J].安徽医药,2016,20(7):1237-1240.
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[8]陈一桢,张文娟,唐兰如,等.和厚朴酚长循环脂质体的制备及药动学研究[J].中草药,2017,48(18):3720-3727.
[9]唐兰如,陈一桢,张文娟,等.Box-Behnken Design-响应面优化法优化PEG化和厚朴酚长循环纳米脂质体处方[J].湖北大学学报:自然科学版,2018,40(4):418-423.
[10]国瑞琪,王秋红,王向涛,等.和厚朴酚脂质体的制备及其体内外抗乳腺癌作用研究[J].药物评价研究,2017,40(1):48-53.
[11]吴德强.和厚朴酚脂质体的制备及表征[J].实用中西医结合临床,2014,14(8):86-88.
[12]Song JM,Anandharaj A,Upadhyaya P,et al.Honokiol suppresses lung tumorigenesis by targeting EGFR and its downstream effectors[J].Oncotarget,2016,7(36):57752-57769.
[13]Gao DQ,Qian S,Ju T.Anticancer activity of Honokiol against lymphoid malignant cells via activation of ROS-JNK and attenuation of Nrf2 and NF-κB[J].J BUON,2016,21(3):673-679.
[14]仇鲸翔,陈俐娟.和厚朴酚衍生物对肝癌细胞(HepG2)凋亡的影响[J].四川化工,2014,17(1):1-3.
[15]张玉碧,张辉,吕林华,等.和厚朴酚对人肝癌Hep G2细胞增殖及NF-k B m RNA表达的影响[J].齐齐哈尔医学院学报,2012,33(12):1562-1563.
[16]董兵轮,丛威亮,李晓明,等.和厚朴酚对人肝癌SMMC-7721细胞体外抗肿瘤活性的研究[J].世界最新医学信息文摘,2016,16(65):3-4.
[2]Nagalingam A,Arbiser JL,Bonner MY,et al.Honokiol activates AMP-activated protein kinase in breast cancer cells via an LKB1-dependent pathway and inhibits breast carcinogenesis[J].Breast Cancer Res,2012,14(1):R35.
[3]郑欢,于鑫,李之韬,等.高载药量和厚朴酚纳米粒的制备及其抗肿瘤作用研究[J].药物评价研究,2015,38(3):292-296.
[4]张斌,谷克仁,潘丽,等.脂质体的性质及制备方法的研究进展[J].粮食与油脂,2015,28(10):17-20.
[5]张宇,白海昕,邵永祥,等.和厚朴酚脂质体联合阿霉素抑制4T1细胞增殖的机制研究[J].中国实验诊断学,2012,16(6):983-985.
[6]王冠杰,赵娅娟,董济民.脂质体和厚朴酚联合热疗对腺样囊性癌增殖的协同抑制作用[J].安徽医药,2016,20(7):1237-1240.
[7]赵呈雷,黄一平,田耀洲,等.HPLC法同时测定桂朴微丸中厚朴酚、和厚朴酚的含量[J].中国药房,2016,27(33):4695-4697.
[8]陈一桢,张文娟,唐兰如,等.和厚朴酚长循环脂质体的制备及药动学研究[J].中草药,2017,48(18):3720-3727.
[9]唐兰如,陈一桢,张文娟,等.Box-Behnken Design-响应面优化法优化PEG化和厚朴酚长循环纳米脂质体处方[J].湖北大学学报:自然科学版,2018,40(4):418-423.
[10]国瑞琪,王秋红,王向涛,等.和厚朴酚脂质体的制备及其体内外抗乳腺癌作用研究[J].药物评价研究,2017,40(1):48-53.
[11]吴德强.和厚朴酚脂质体的制备及表征[J].实用中西医结合临床,2014,14(8):86-88.
[12]Song JM,Anandharaj A,Upadhyaya P,et al.Honokiol suppresses lung tumorigenesis by targeting EGFR and its downstream effectors[J].Oncotarget,2016,7(36):57752-57769.
[13]Gao DQ,Qian S,Ju T.Anticancer activity of Honokiol against lymphoid malignant cells via activation of ROS-JNK and attenuation of Nrf2 and NF-κB[J].J BUON,2016,21(3):673-679.
[14]仇鲸翔,陈俐娟.和厚朴酚衍生物对肝癌细胞(HepG2)凋亡的影响[J].四川化工,2014,17(1):1-3.
[15]张玉碧,张辉,吕林华,等.和厚朴酚对人肝癌Hep G2细胞增殖及NF-k B m RNA表达的影响[J].齐齐哈尔医学院学报,2012,33(12):1562-1563.
[16]董兵轮,丛威亮,李晓明,等.和厚朴酚对人肝癌SMMC-7721细胞体外抗肿瘤活性的研究[J].世界最新医学信息文摘,2016,16(65):3-4.