HepG2.2.15细胞对同时包载丁香苦苷和羟基酪醇纳米粒摄取机制研究.docVIP

HepG2.2.15细胞对同时包载丁香苦苷和羟基酪醇纳米粒摄取机制研究 　　摘要：目的 考察HepG2.2.15细胞对同时包载丁香苦苷和羟基酪醇纳米粒（nanoparticles co-loaded with syringopicroside and hydroxytyrosol，SH-NPs）的摄取机制。方法 采用沉淀法制备SH-NPs，以异硫氰酸荧光素为荧光标记物，采用流式细胞仪研究HepG2.2.15细胞对SH-NPs的摄取机制。结果 秋水仙素为抑制剂，孵育时间在0.5～24 h范围内，阳性细胞百分数由1.9%增加到56.4%；药物浓度为125、250、500 ?g/mL时，阳性细胞百分数分别为4.9%、3.4%、3.9%。氯喹为抑制剂，孵育时间在0.5～24 h范围内，阳性细胞百分数由7.4%增加到55.4%；药物浓度为125、250、500 ?g/mL时，阳性细胞百分数分别为19.5%、22.5%、27.6%。结论 秋水仙素与氯喹对HepG2.2.15细胞摄取有抑制作用，且HepG2.2.15细胞对SH-NPs的摄取与药物浓度、孵育时间呈正相关，推断HepG2.2.15细胞对SH-NPs细胞的摄取机制为非特异性吸附内吞。 　　关键词：丁香苦苷；羟基酪醇；聚乙二醇-聚乳酸乙醇酸共聚物纳米粒；HepG2.2.15细胞；摄取机制 　　DOI：10.3969/j.issn.1005-5304.2018.03.018 　　中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2018）03-0081-05 　　Abstract： Objective To investigate the uptake mechanism of HepG2.2.15 cells to the nanoparticles co-loaded with syringopicroside and hydroxytyrosol （SH-NPs）. Methods The nanoparticles were prepared by using a nanoprecipitation method with mPEG-PLGA as nano-carrier co-loaded with syringopicroside and hydroxytyrosol. The uptake mechanism of HepG2.2.15 cells to SH-NPs was studied by fluorescence microscopy and flow cytometry using fluoresceineisothiocyanate （FITC） as a fluorescent marker. Results With colchicine as the inhibitor， the incubation time ranged from 0.5 to 24 h， the percentage of positive cells increased from 1.9% to 56.4%； When the drug concentration was 125， 250 ?g/mL and 500 ?g/mL， the positive cell percentages were 4.9%， 3.4% and 3.9%. With chloroquine as the inhibitor； the incubation time ranged from 0.5 to 24 h， the percentage of positive cells increased from 7.4% to 55.4%； When the drug concentration was 125， 250 and 500 ?g/mL， the percentage of positive cells was 19.5%， 22.5% and 27.6%. Conclusion Colchicine and chloroquine have an inhibitory effect on HepG2.2.15 cells uptake， and the uptake of SH-NPs in HepG2.2.15 cells was positively correlated with drug concentration and incubation time. It can be concluded that the uptake mechanism of HepG2.2.15 cells to SH-NPs was nonspecific