负载姜黄素的工程化细胞膜纳米颗粒的制备及其用于小鼠乳腺癌治疗的研究  

Preparation of curcumin-loaded engineered cell membrane nanoparticles and its impact on breast cancer treatment in mice

作　　者：廖谊芳 饶浪 张云娇 LIAO Yifang;RAO Lang;ZHANG Yunjiao(School of Medicine,South China University of Technology,Guangzhou 510006,China;Institute of Biomedical Health Technology and Engineering,Shenzhen Bay Laboratory,Shenzhen 518132,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]深圳湾实验室,医药健康技术与工程研究所,广东深圳518132

出　　处：《中国病理生理杂志》2024年第7期1173-1181,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金优秀青年基金(No.T2222014)。

摘　　要：目的:制备负载姜黄素(Cur)的工程化细胞膜仿生纳米颗粒(PD1-Cur@PLGA NPs),探讨其对小鼠乳腺癌的治疗效果及潜在的肿瘤免疫调控作用。方法:构建过表达程序性死亡受体1(PD1)的工程化小鼠乳腺癌4T1-PD1细胞,并通过流式细胞术分析PD1表达率。提取4T1-PD1细胞膜,通过冰浴超声将其涂覆在负载Cur的聚乳酸-羟基乙酸共聚物(PLGA)纳米颗粒(Cur@PLGA NPs)表面以制备PD1-Cur@PLGA NPs;紫外可见分光光度计检测各纳米制剂的Cur负载;动态光散射和透射电镜分析各纳米制剂的粒径和形貌。将4T1细胞分为阴性对照(PLGA NPs和PD1-NVs)组、实验组(PD1-Cur@PLGA NPs组)、平行对照(Cur@PLGA NPs)组及阳性对照(Cur)组;CCK-8法检测各组细胞活力;流式细胞术分析各组细胞凋亡水平。将4T1细胞移植瘤小鼠随机分为PBS组、PD1-NVs组及PD1-Cur@PLGA NPs组,进行体内治疗实验;治疗结束后,组织染色观察主要器官的病理变化,检测肿瘤增殖(Ki67)、凋亡(TUNEL)、CD4^(+)和CD8^(+)T细胞的浸润和活性指标。结果:(1)4T1-PD1细胞系PD1表达率高达78%;(2)PD1-Cur@PLGA NPs呈类细胞壳核结构且粒径为100~200 nm;(3)PD1-Cur@PLGA NPs提高了Cur的生物相容性,且能有效诱导4T1细胞凋亡;(4)与对照组相比,PD1-Cur@PLGA NPs显著抑制了4T1乳腺癌的进展(P<0.01),且安全性高;肿瘤组织的Ki67阳性表达降低,细胞凋亡显著,CD4^(+)和CD8^(+)T细胞的浸润和活性增强。结论:PD1-Cur@PLGA NPs提高了Cur的生物相容性,并对小鼠乳腺癌细胞具有杀伤作用。该组合制剂在体内展现出良好的治疗效果、安全性和潜在的抗肿瘤免疫调控作用。AIM:To investigate the therapeutic efficacy of curcumin(Cur)-loaded engineered cell membrane mimetic nanoparticles(PD1-Cur@PLGA NPs)in treating breast cancer in mice,and to explore their tumor immunomodulatory effects.METHODS:Engineered mouse breast cancer 4T1-PD1 cell line expressing programmed death 1(PD1)was established,and PD1 expression level was analyzed by flow cytometry.The 4T1-PD1 cell membranes were extracted and coated on the surface of Cur-loaded poly(lactic-co-glycolic)acid(PLGA)nanoparticles(Cur@PLGA NPs)using ice bath sonication to obtain PD1-Cur@PLGA NPs.The Cur loaded in various NPs was detected using UV-visible spectrophotometry.Particle size and morphology were analyzed by using dynamic light scattering and transmission electron microscopy.The 4T1 cells were divided into negative control(PLGA NPs and PD1-NVs),experimental(PD1-Cur@PLGA NPs),parallel control(Cur@PLGA NPs),and positive control(Cur)groups.In each group,cell viability was assessed by CCK-8 assay,and cell apoptosis was determined through flow cytometry.To perform treatment experiments in vivo,4T1 cellbearing tumor mice were randomly divided into PBS,PD1-NVs,and PD1-Cur@PLGA NPs groups.At the end of treatments,tissues of major organs were stained to detect pathological changes,as well as indicators of tumor proliferation(Ki67),apoptosis(TUNEL),and infiltration and activity of T cells(CD4^(+)and CD8^(+))in tumor tissues.RESULTS:The PD1 expression in 4T1-PD1 cell lines reached 78%.PD1-Cur@PLGA NPs exhibited a core-shell structure with particle sizes ranging from 100 to 200 nm.PD1-Cur@PLGA NPs enhanced the biocompatibility compared to free Cur and exhibited a strong apoptosis-inducing effect on 4T1 cells.Compared with control group,PD1-Cur@PLGA NPs significantly inhibited 4T1 breast tumor growth in vivo(P<0.01),without apparent toxic side effects.Treatment with PD1-Cur@PLGA NPs reduced Ki67 expression,increased cell apoptosis,and enhanced infiltration and activity of CD4^(+)and CD8^(+)T cells in tumor tissues.CONCLUSION:PD1-Cur@PLGA NPs

关 键 词：细胞膜生物材料 药物递送 姜黄素 乳腺癌 

分 类 号：R363[医药卫生—病理学] R737.9[医药卫生—基础医学] R730.23