血小板膜修饰过氧化氢酶/二氧化硅纳米粒抑制机体辐射感染  

Platelet membrane-modified catalase/silica nanoparticles inhibit radiation infection

作　　者：熊太农 李陈文娅 陈银 韩松伶 王成[1,2] 王军平[1,2] XIONG Tainong;LI Chenwenya;CHEN Yin;HAN Songling;WANG Cheng;WANG Junping(Institute of Combined Injury,Faculty of Military Preventive Medicine,Army Medical University(Third Military Medical University),Chongqing,China;State Key Laboratory of Trauma and Chemical Poisoning,Faculty of Military Preventive Medicine,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)军事预防医学系复合伤研究所,重庆 [2]陆军军医大学(第三军医大学)创伤与化学中毒全国重点实验室,重庆

出　　处：《陆军军医大学学报》2025年第6期602-612,共11页Journal of Army Medical University

基　　金：国家自然科学基金重点项目(81930090);创伤与化学中毒全国重点实验室自定课题(SKLYQ202101)。

摘　　要：目的通过制备能够靶向白细胞的血小板膜修饰过氧化氢酶/二氧化硅纳米粒(platelet membrane modified catalase/silica nanoparticles,PCNP),为辐射感染并发症的防治提供有效策略。方法利用血小板膜、过氧化氢酶(catalase,CAT)以及二氧化硅制备PCNP以及过氧化氢酶/二氧化硅纳米粒(catalase/silica nanoparticles,CNP),并通过细胞存活实验、溶血实验以及小鼠尾静脉给药后急性毒性实验初步评估PCNP生物安全性;培养基、FITC标记的PCNP(FITC^(+)PCNP)及FITC标记的CNP(FITC^(+)CNP)分别与人外周血B淋巴细胞(AHH-1)和小鼠单核巨噬细胞(RAW264.7)共孵育,分为对照组、FITC^(+)PCNP组及FITC^(+)CNP组,利用激光共聚焦观察细胞内荧光强度来评价PCNP的白细胞靶向功能;将AHH-1分为对照组、辐照组、血小板膜组、CNP(100μg/mL)组和PCNP(100μg/mL)组,与培养基、培养基、血小板膜混悬液、CNP或PCNP溶液共孵育后进行6 Gy Co~(60)γ射线辐照,通过检测羟自由基等活性氧自由基(reactive oxygen species,ROS)水平及细胞凋亡情况评估纳米粒体外减轻白细胞放射损伤的作用;20只C57BL/6雄性小鼠(体质量18~20 g)简单随机抽样分为(n=10):辐照组与10mg/kg PCNP组,小鼠尾静脉注射生理盐水或PCNP后2 h时进行5 Gy Co~(60)γ射线全身辐照,辐照后2 h时再腹腔注射多重耐药鲍曼不动杆菌(multidrug-resistant Acinetobacter baumannii MDR-AB),通过检测主要脏器细菌载量评估纳米药物抑制辐照后感染效应。结果PCNP水合粒径为91.3 nm,其在低于400μg/mL的浓度下未表现出明显的细胞毒性及溶血毒性;小鼠尾静脉注射PCNP(20mg/kg)后体质量正常增长,心、肝、脾、肺、肾等主要脏器组织未出现病理改变;在AHH-1和RAW264.7细胞中,与FITC^(+)CNP组相比,PCNP的靶向性都具有显著优势[(15.45±3.48)%vs(9.33±2.03)%,P<0.01;(11.25±2.08)%vs(7.06±0.71)%,P<0.001];细胞防护能力检测实验中,与辐照组相比,PCNP能有效降低AHH-1细胞ROObjective To provide an effective strategy for the prevention and treatment of radiation-induced infections by preparing platelet membrane-modified catalase/silica nanoparticles(PCNP)capable of targeting leukocytes.Methods PCNP and catalase/silica nanoparticles(CNP)were prepared by using platelet membrane,catalase(CAT)and silica,and its biological safety was preliminarily evaluated with cell survival test,hemolysis test and acute toxicity test in mice after tail vein administration;The culture medium,FITC labeled(FITC^(+) )PCNP and FITC labeled(FITC^(+) )CNP were co-incubated with human peripheral blood B lymphocytes(AHH-1)and mouse monocyte macrophages(RAW264.7),respectively.Thus,there were control group,FITC^(+) PCNP group and FITC^(+) CNP group of AHH-1 and RAW264.7 cells.Laser confocal microscopy was used to observe the intracellular fluorescence intensity of PCNP to evaluate the leukocytes targeting function.AHH-1 cells were divided into control,irradiation,platelet membrane,CNP(100µg/mL)and PCNP(100µg/mL)groups.After corresponding co-incubation,the cell media were exposed to6 Gy Co60γirradiation.The generation of reactive oxygen species(ROS)and cell apoptosis were measured to determine the effect of nanoparticles on reducing radiation injury of leukocytes.Twenty C57BL/6 male mice(weighing 18~20 g)were randomly divided into irradiation group(n=10)and 10 mg/kg PCNP group(n=10).In 2 h after corresponding agents were injected into the mice through tail vein,the mice received whole-body irradiation of 5 Gy Co60γray,and then in 2 h later,they were given intraperitoneal injection of multidrug resistant Acinetobacter baumannii(MDR-AB).The infection inhibitory effect of PCNP after irradiation was evaluated by detecting the bacterial load in main organs.Results The hydration particle of PCNP is 91.3 nm in size,and does not exhibit significant cytotoxicity or hemolytic toxicity at concentrations<400µg/mL.Intravenous injection of 20 mg/kg PCNP resulted in normal increase in the body weight but no obvious pathologi

关 键 词：辐射感染 过氧化氢酶 白细胞靶向 血小板膜修饰纳米粒 

分 类 号：R345[医药卫生—基础医学] R818.74R945