携载miRNA-124纳米粒子经鼻吸入对缺血性脑卒中模型大鼠的神经保护作用  

Neuroprotective effect of nasal inhalation of miRNA-124 nanoparticles on ischemic stroke model rats

作　　者：郝如彬 王浩天[2] 杨丽华[3] 马春[4] 李淑玲[4] 李欣欣 李铁术 HAO Rubin;WANG Haotian;YANG Lihua;MA Chun;Li Shuling;LI Xinxin;LI Tieshu(Department of Chinese Internal Medicine,School of Traditional Chinese Medicine,Changchun University of Chinese Medicine,Changchun130117,China;Department of Microbiology and Biochemical Pharmacy,Institute of Biopharmaceutics,School of Pharmacy,Jilin University,Changchun130021,China;Institute for Geriatrics,Affiliated Hospital,Changchun University of Chinese Medicine,Changchun130021,China;Department of Geriatrics,Affiliated Hospital,Changchun University of Chinese Medicine,Changchun130021,China)

机构地区：[1]长春中医药大学中医学院中医内科学教研室,吉林长春130117 [2]吉林大学药学院生物药学系微生物与生化药学教研室,吉林长春130021 [3]长春中医药大学附属医院老年病研究所,吉林长春130021 [4]长春中医药大学附属医院老年病科,吉林长春130021

出　　处：《吉林大学学报（医学版）》2020年第6期1208-1214,I0006,共8页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科学计划支撑项目资助课题(20140204045YY);吉林省财政厅省级产业创新专项资金项目资助课题(2018C049-3)。

摘　　要：目的:探讨携载微小RNA124(miRNA-124)纳米粒子经鼻吸入对缺血性脑卒中模型大鼠脑内药物分布、神经功能评分、脑组织含水量及血清炎性因子的药效学影响,阐明其对缺血性脑卒中大鼠的神经保护作用。方法:将靶向物RVG29偶联在聚乙二醇‐聚乳酸‐羟基乙酸共聚物(PEG-PLGA)纳米粒子上,观察所制备纳米粒子的粒径和包封率,通过向大鼠施用载有近红外染料1,1'-双十八烷基-3,3,3,3-四甲基吲哚三碳花菁碘化物(DiR),并给予RVG29修饰的PEG-PLGA纳米粒子,分析经鼻吸入纳米粒子后不同时间药物的生物分布。采用线栓法建立脑缺血再灌注大鼠模型,90只大鼠随机分为假手术组、模型组、法舒地尔组、miRNA-124经鼻吸入给药组、PEG-PLGA/miRNA-124经鼻吸入给药组和RVG29-PEG-PLGA/miRNA-124经鼻吸入给药组,每组6只。检测各组大鼠神经功能评分和脑组织含水量,ELISA法检测各组大鼠外周血清中白细胞介素4(IL-4)、白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)和白细胞介素1β(IL-1β)水平。结果:可鼻吸入脑的RVG29-PEG-PLGA纳米粒子的平均粒径为204 nm,包封率为28%。假手术组大鼠无神经功能障碍;与假手术组比较,模型组大鼠神经学评分和脑组织含水量,血清中IL-4、IL-6、TNF-α和IL-1β水平升高(P<0.05);与模型组比较,法舒地尔组、miRNA-124经鼻吸入给药组、PEG-PLGA/miRNA-124经鼻吸入给药组和RVG29-PEG-PLGA/miRNA-124经鼻吸入给药组大鼠神经学评分,脑组织含水量,血清中IL-4、IL-6、TNF-α和IL-1β水平降低(P<0.05);与法舒地尔组比较,RVG29-PEG-PLGA/miRNA-124经鼻吸入给药组大鼠神经学评分和脑组织含水量,血清中IL-4、IL-6、TNF-α和IL-1β水平降低(P<0.05)。结论:RVG29-PEG-PLGA纳米粒子具有较好的经鼻吸入脑能力。对缺血性脑卒中大鼠进行纳米粒子携载miRNA-124干预可以改善大鼠神经功能障碍、减轻脑水肿以及改善炎症,起到神经保护作用。Objective:To investigate the pharmacodynamic effects of nasal inhalation of microRNA-124(miRNA-124)nanoparticles on the durg distribution in brain tissue,neurological function score,water content in brain tissue and levels of serum inflammatory factors in the ischemic stroke model rats,and to elucidate its neuroprotective effect on the rats with ischemia stroke.Methods:The target RVG29 was coupled to the poly(ethylene glycol)-poly(lactic-coglycolic acid)(PEG-PLGA)nanoparticles,the particle size and encapsulation efficiency of the prepared nanoparticles were measured.The PEG-PLGA nanoparticles modified with neurotropic virus-derived peptide RVG29 labeled nearinfrared dye 1,1'-dioctadecyl-3,3,3,3-tetramethylindole three-carbon cyanine iodide(DiR)were given to the rats,and the durg biodistribution at different time points after nasal inhalation was measured.The rat model of cerebral ischemiareperfusion was established by the suture method,and 90 rats were randomly divided into sham operation group,model group,Fasudil group,miRNA-124 nasal inhalation group,PEG-PLGA/miRNA-124 nasal inhalation group,and RVG29-PEG-PLGA/miRNA-124 nasal inhalation group,and there were 6 rats in each group.The neurological function scores and water contents in brain tissue of the rats in various groups were evaluated;the levels of interleukin-4(IL-4),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and interleukin-1β(IL-1β)in peripheral serum of the rats in various groups were detected by ELISA method.Results:The average particle size of RVG29-PEG-PLGA nanoparticles was 204 nm,and the encapsulation efficiency was 28%.The rats in sham operation group had no neurological dysfunction.Compared with sham operation group,the neurological function score,water content in brain tissue,and levels of serum IL-4,IL-6,TNF-α,and IL-1βof the rats in model group were increased(P<0.05).Compared with model group,the neurological function scores,water contents in brain tissue,and levels of serum IL-4,IL-6,TNF-α,and IL-1βof the rats in Fasudil group

关 键 词：纳米粒子 脑缺血再灌注 神经保护 经鼻吸入 RVG29 

分 类 号：R743.[医药卫生—神经病学与精神病学]