活性氧清除性Ce6-鲁米诺脂质体的制备及其对小鼠放射性肺炎的保护作用研究  

Preparation of reactive oxygen species scavenging Ce6-Luminol encapsulated liposomes and their protective effect against radiation pneumonitis in mice

作　　者：雷航科 黄定德 LEI Hangke;HUANG Dingde(Department of Nuclear Medicine,First Affiliated Hospital,Army Medical University(Third Military Medical University),Chongqing,China)

机构地区：[1]陆军军医大学(第三军医大学)第一附属医院核医学科,重庆

出　　处：《陆军军医大学学报》2025年第6期489-497,共9页Journal of Army Medical University

基　　金：重庆市自然科学基金面上项目(CSTB2022NSCQ-MSX0150)。

摘　　要：目的制备活性氧清除性Ce6-鲁米诺脂质体(Ce6-Luminol encapsulated liposomes,CLL),探究基于纳米材料的活性氧清除策略对小鼠放射性肺炎是否具有保护作用。方法化学合成活性氧清除性材料Ce6-鲁米诺,通过脂质体技术将其包封得到活性氧清除性CLL;用激光粒度仪对CLL的粒径、多分散指数及Zeta电位进行表征;流式细胞术检测A549细胞、HUVEC细胞及Raw 264.7细胞对CLL的摄取;CCK-8法评估摄取CLL后对细胞活力的影响;活体成像系统考察CLL在放射性肺炎小鼠肺组织的分布情况;将75只雄性C57BL/6J小鼠(6~8周龄,17~19 g)按体质量分层抽样,分为正常对照组(Normal),0、2、10和50 mg/kg CLL干预组,每组15只。CLL干预组小鼠胸部接受15 Gy X射线辐照,于辐照后第0.25、3及5天尾静脉注射相应剂量CLL。辐照后第7天检测小鼠肺组织活性氧H_(2)O_(2)含量,炎症因子TNF-α、IL-1β和炎症细胞趋化因子MCP-1、KC的含量,以及支气管肺泡灌洗液中性粒细胞比例,评价CLL对小鼠放射性肺炎的干预效果。结果成功合成了活性氧清除性材料Ce6-鲁米诺,制备的CLL平均粒径为(127.8±0.5)nm,多分散指数为0.164±0.005,Zeta电位为(-23.6±1.4)mV。CLL可被A549细胞、HUVEC细胞及Raw 264.7细胞以时间及剂量依赖的方式所摄取,细胞活力均未见剧烈降低。活体成像发现,与正常小鼠相比,CLL在放射性肺炎小鼠肺组织的分布显著增多(F=351.741,P<0.05)。CLL干预可显著降低放射性肺炎小鼠肺组织活性氧H_(2)O_(2)(F=15.183,P<0.05)、炎症因子TNF-α(F=5.150,P<0.05)和IL-1β(F=10.036,P<0.05)、炎症细胞趋化因子MCP-1(F=3.777,P<0.05)和KC(F=3.755,P<0.05)的含量,以及支气管肺泡灌洗液中性粒细胞比例(F=17.751,P<0.05)。结论活性氧清除性脂质体CLL可减轻小鼠放射性肺炎,基于纳米材料的活性氧清除策略对小鼠放射性肺炎具有保护作用。Objective To prepare reactive oxygen species(ROS)scavenging Ce6-Luminol encapsulated liposomes(CLL)and to explore the protective effect of CLL against radiation pneumonitis in mice.Methods ROS scavenging material Ce6-Luminol was chemically synthesized,and then was encapsulated using liposome technology to prepare ROS scavenging CLL liposomes.A laser particle size analyzer was used to characterize particle size,polydispersity index,and zeta potential of prepared CLL liposomes.Flow cytometry was employed to detect the uptake of CLL by A549 cells,HUVEC and Raw 264.7 cells,respectively,and CCK-8 assay was utilized to evaluate the cell viability after CLL uptake.The distribution of CLL in the lung tissue of mouse model of radiation lung injury was investigated using In Vivo Imaging System(IVIS).A total of 75 male C57BL/6J mice(6~8 weeks old,17~19 g)were randomly divided into a normal control group(Normal)and CLL treatment groups(0,2,10,and 50 mg/kg),with 15 animal in each group.After the mice in the CLL treatment groups received 15 Gy X-ray irradiation on their chests,they received CLL injection at the corresponding through tail vein on days 0.25,3,and 5 post-irradiation.Then on day 7 post-irradiation,the contents of H2O2,inflammatory cytokines TNF-αand IL-1β,and chemokines monocyte chemoattractant protein(MCP-1)and keratinocyte chemoattractant(KC)in lung tissue,and the proportion of neutrophils in bronchoalveolar lavage fluid(BALF)were measured to evaluate the efficacy of CLL in treatment of radiation pneumonitis in mice.Results The ROS scavenging material Ce6-Luminol was successfully synthesized,and CLL liposomes were prepared,with an average particle size of 127.8±0.5 nm,a polydispersity index of 0.164±0.005,and a Zeta potential of-23.6±1.4 mV.The prepared CLL liposomes were taken up by A549 cells,HUVEC and Raw264.7 cells in a time-and dose-dependent manner,without a sharply decrease in cell viability.IVIS revealed that there were significantly more CLL liposomes distributed in the lung tissues of radiation pn

关 键 词：放射性肺炎 电离辐射 活性氧 炎症 活性氧清除性脂质体 

分 类 号：R563.1[医药卫生—呼吸系统] R818.84[医药卫生—内科学] R945[医药卫生—临床医学]