负载双氢青蒿素的中空介孔二氧化锰纳米颗粒对胰腺癌BxPC-3细胞的杀伤效应  

Killing effect of hollow mesoporous MnO_(2) nanoparticles loaded with dihydroartemisinin on pancreatic cancer BxPC-3 cell line

作　　者：刘刚 刘明宇 李修靖 余跃 LIU Gang;LIU Mingyu;LI Xiujing;YU Yue(Division of Gastroenterology,the First Affiliated Hospital of Anhui University of Chinese Medicine,Hefei 230031,China;Division of Gastroenterology,the First Affiliated Hospital of USTC,Division of Life Science and Medicine,University of Science and Tech-nology of China,Hefei 230001,China)

机构地区：[1]安徽中医药大学第一附属医院脾胃科,安徽合肥230031 [2]中国科学技术大学附属第一医院(安徽省立医院)消化科,安徽合肥230001

出　　处：《安徽医学》2023年第6期623-628,共6页Anhui Medical Journal

基　　金：国家自然科学基金面上项目(编号:31870993)。

摘　　要：目的探究负载双氢青蒿素(DHA)的中空介孔二氧化锰纳米颗粒对胰腺癌BxPC-3细胞的杀伤效应。方法利用二氧化硅模板法合成了纳米中空介孔二氧化锰,通过DHA及聚乙二醇-2000(PEG-2000)共孵育后合成了PEG@MnO_(2)-DHA和未载药的PEG@MnO_(2),并对其进行表征和催化性能验证。利用激光共聚焦显微镜(CLSM)分别检测0、1、2和4 h内BxPC-3细胞对纳米颗粒的摄取情况。采用酶联免疫吸附试验(ELISA)检测不同氧浓度下经PEG@MnO_(2)-DHA处理后BxPC-3细胞内的缺氧诱导因子-1α(HIF-1α)水平,以探究PEG@MnO_(2)-DHA改善肿瘤微环境乏氧情况。最后用噻唑蓝比色法(MTT)和活死细胞染色法观察纳米颗粒对BxPC-3细胞的生长抑制作用。结果本研究成功制备出粒径约为105 nm的PEG@MnO_(2)-DHA,并且其能够在RPMI-1640培养基(含10%胎牛血清)中稳定保存。CLSM结果显示,PEG@MnO_(2)-DHA在1h后就能够被BxPC-3细胞有效摄取。ELISA结果显示,低氧条件下,使用20μg/mL的PEG@MnO_(2)-DHA处理后的BxPC-3细胞内HIF-1α水平与使用磷酸缓冲盐溶液(PBS)处理后的BxPC-3细胞内HIF-1α水平相比明显下降(P<0.05),略低于常氧条件下PBS处理后的BxPC-3细胞中HIF-1α水平(P=0.846)。MTT结果显示,在DHA浓度为15μg/mL时,PEG@MnO_(2)-DHA处理组细胞存活率仅为21.81%,低于游离DHA处理组的46.03%(P<0.05)。结论负载DHA之后的中空介孔二氧化锰纳米颗粒可以被BxPC-3细胞有效摄取,并与释放出来的DHA反应产生活性氧,同时能改善肿瘤微环境乏氧情况,增强对胰腺癌细胞的杀伤效应。Objective To investigate the killing effect of hollow mesoporous MnO_(2)nanoparticlesloaded withdihydroartemisinin(DHA)on pancreatic cancer BxPC-3 cells.Methods Hollow mesoporous MnO_(2)nanoparticles were synthesized by SiO2template method.PEG@MnO_(2)-DHA and PEG@MnO_(2)were synthesized by co-incubation with DHA and Polyethylene glycol 2000(PEG-2000),then characteristics and catalytic properties were investigated.The uptake of nanoparticles by BxPC-3 cells at different time(0,1,2 and 4 h)was detected by laser confocal microscopy(CLSM).The level of HIF-1αin BxPC-3 cells was detected by ELISA assay to investigate the effect of PEG@MnO_(2)-DHA on hypoxia in tumour microenvironment(TME).Lastly,the growth inhibition of BxPC-3 cells by nanoparticles was observed by MTT assay and cell death staining.Results PEG@MnO_(2)-DHA with a particle size of about 105 nm was successfully prepared and could be stably stored in RPMI-1640 medium containing 10%fetal bovine serum.CLSM results showed that PEG@MnO_(2)-DHA could be efficiently taken up by BxPC-3 cells.ELISA results showed that HIF-1αlevels in BxPC-3 cells cultured under hypoxic conditions were significantly reduced after treatment with PEG@MnO_(2)-DHA at 20μg/mL compared with PBS treatment(P<0.05),and the HIF-1αlevel in BxPC-3 cells was slightly lower than that in the normoxia group after PBS treatment(P=0.846).MTT results showed that the cell survival rate of the PEG@MnO_(2)-DHA treated group was only 21.81%at 15μg/mL DHA concentration,which was lower than46.03%of the free DHA treated group(P<0.05).Con⁃clusions The hollow mesoporous MnO_(2)nanoparticles loaded with DHA can be effectively taken up by BxPC-3 cellsand react with the released DHA to produce reactive oxygen species,which can also improve the lack of oxygen in the tumor microenvironment and enhance the killing effect on pancreatic cancer cells.

关 键 词：双氢青蒿素 中空介孔二氧化锰 活性氧 胰腺癌 

分 类 号：R735.9[医药卫生—肿瘤]