齐墩果酸聚乙二醇修饰自组装及其对白血病细胞K562的增殖抑制作用  

Self-assembly of polyethylene glycol modified oleanolic acid and its inhibitory effect on K562 cell proliferation

作　　者：赵彬男 毛晶 葛堂栋[1] 张鹏霞[1] 王迪迪[1] 朴金花[1] ZHAO Binnan;MAO Jing;GE Tangdong;ZHANG Pengxia;WANG Didi;PIAO Jinhua(Department of Biochemistry and Molecular Biology,Basic Medical College of Jiamusi University,Jiamusi 154007,China;Department of Medical Cosmetology,Yantai Affiliated Hospital of Binzhou Medical University)

机构地区：[1]佳木斯大学基础医学院生物化学与分子生物学教研室,佳木斯154007 [2]滨州医学院烟台附属医院医学美容科

出　　处：《山西医科大学学报》2022年第10期1227-1234,共8页Journal of Shanxi Medical University

基　　金：黑龙江省北药与功能食品优势特色学科建设项目(2018-TSXK-02)。

摘　　要：目的用聚乙二醇(polyethylene glycol,PEG)对齐墩果酸(oleanolic acid,OA)进行修饰,并探讨其衍生物对人慢性粒细胞白血病细胞K562的抑制作用。方法以OA和PEG为原料,酯化法、纳米沉淀法合成齐墩果酸聚乙二醇衍生物,核磁共振、电子显微镜、动态光散射检测衍生物表征。CCK-8法检测齐墩果酸聚乙二醇衍生物对K562细胞增殖抑制的影响,倒置显微镜检测齐墩果酸聚乙二醇衍生物对K562细胞形态的影响,流式细胞术检测齐墩果酸聚乙二醇衍生物对K562细胞凋亡的影响,Western blot检测K562细胞凋亡蛋白(Caspase-3、Caspase-9、PARP-1)及其剪切后蛋白表达。结果成功合成了齐墩果酸聚乙二醇自组装颗粒OAP5,粒径为(584.49±76.2)nm,结构为核-壳结构,电位趋近于0 mV。OAP5对白血病细胞K562具有增殖抑制作用,处理48 h后的IC_(50)为(16.10±0.64)μg/ml。OAP5处理K562细胞后,细胞密度降低,萎缩变形,折光性变差。OAP5能够促进K562细胞凋亡,剪切Caspase-3、剪切Caspase-9、剪切PARP-1表达增多(P<0.05)。结论OAP5能够通过诱导白血病细胞K562凋亡作用而抑制K562细胞增殖,为齐墩果酸抗肿瘤临床应用提供了依据。Objective To modify the oleanolic acid(OA)with polyethylene glycol(PEG),and explore the inhibitory effect of its derivatives on human chronic myeloid leukemia cell K562.Methods Oleanolic acid polyethylene glycol derivatives were synthesized from OA and PEG by esterification and nano-precipitation method.The derivatives were characterized by NMR,electron microscope and dynamic light scattering.The effects of derivatives on K562 cell proliferation were detected by CCK-8 method.The effect of polyethylene glycol derivatives on K562 cell morphology was observed by inverted microscopy.Apoptosis was detected by flow cytometry.The expression of apoptotic proteins(Caspase-3,Caspase-9,and PARP-1)and their cleaved proteins in K562 cells was determined by Western blot.Results Oleanolic acid polyethylene glycol self-assembled particle OAP5 was successfully synthesized,with a particle size of(584.49±76.2)nm,a core-shell structure and a potential of 0 mV.OAP5 had a inhibitory effect on the proliferation of leukemic cells K562,and the IC_(50) was(16.10±0.64)μg/ml after treatment with OAP5 for 24 h.The density of K562 cells was decreased,the morphology was atrophic and deformation,and the refraction became worse after OAP5 treatment.OAP5 promoted the apoptosis of K562 cells,and increased the expression of cleaved Caspase-3,cleaved Caspase-9 and cleaved PARP-1(P<0.05).Conclusion OAP5 can inhibit the proliferation of K562 cells by inducing the apoptosis of K562 cells,which provides a basis in the clinical application of oleanolic acid for anti-tumor.

关 键 词：齐墩果酸 聚乙二醇 自组装 K562 凋亡 

分 类 号：R733.7[医药卫生—肿瘤]