活性氧响应靶向纳米粒对细胞氧化应激和细胞炎症因子的影响  

Effects of reactive oxygen species-responsive targeted nanoparticles on cellular oxidative stress and inflammatory cytokines

作　　者：曹隆檬 金虎[1] Cao Longmeng;Jin Hu(Department of Cardiovascular,Wenzhou TCM Hospital,Wenzhou 325000,China)

机构地区：[1]浙江省温州市中医院心血管科,325000

出　　处：《心脑血管病防治》2024年第5期19-22,共4页CARDIO-CEREBROVASCULAR DISEASE PREVENTION AND TREATMENT

基　　金：温州市科技局课题(Y20210895)。

摘　　要：目的探讨活性氧(ROS)响应靶向纳米粒(OXb CD NP)对细胞氧化应激和细胞炎症因子的影响。方法采用化学键合方式自制ROS响应靶向纳米粒。(1)选用RAW264.7细胞,随机分为四组,空白1组(无任何处理),对照1组(PMA刺激),10μg/m L OXbCD NP1组(10μg/m L OXbCD NP孵育后再加入PMA刺激),100μg/mL OXbCD NP1组(100μg/mL OXbCD NP孵育后再加入PMA刺激)。用DCFH-DA探针染色,流式细胞术检测细胞内的ROS水平,并荧光成像;(2)选用RAW264.7细胞,随机分为四组,空白2组(无任何处理),对照2组(IFN-γ和LPS刺激),10μg/mL OXbCD NP 2组(10μg/mL OXbCD NP孵育后加入IFN-γ和LPS刺激),100μg/mL OXbCD NP 2组(100μg/mL OXbCD NP孵育后加入IFN-γ和LPS刺激),用ELISA法检测白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)含量;(3)将RAW264.7细胞用0μg/m L、5μg/m L、10μg/mL、20μg/mL、40μg/mL、80μg/mL、160μg/mL、320μg/mL、640μg/mL、1280μg/mL OXb CD NP培养液孵育12 h或24 h后,用MTT法检测细胞存活率。结果(1)相比于对照1组,10μg/m L和100μg/m L OXbCD NP 1组ROS水平下降,荧光强度下降,差异有统计学意义(P<0.01);(2)与对照2组比较,10μg/m L和100μg/m L OXbCD NP 2组IL-1β、TNF-α水平下降,差异有统计学意义(P<0.01)。(3)纳米粒与细胞共孵育12 h,与OXbCD NP水平为0μg/mL(不含OXbCD NP)对比,OXbCD NP水平达到640μg/mL及1280μg/mL时,存活率差异有统计学意义(P<0.05);当孵育时间为24 h,与不含OXbCD NP对比,OXbCD NP水平为320μg/mL、640μg/mL及1280μg/mL时,存活率差异有统计学意义(P<0.05)。结论ROS响应靶向纳米粒在细胞水平具有良好的安全性,并且有抗细胞氧化应激和抗炎症因子反应的作用。Objective To investigate the effects of reactive oxygen species(ROS)-responsive targeted nanoparticles(OXbCD NP)on cellular oxidative stress and inflammatory cytokines.Methods ROSresponsive targeted nanoparticles were synthesized using chemical bonding.(1)RAW264.7 cells were randomly divided into 4 groups:blank 1 group(no treatment),control 1 group(PMA stimulation),10μg/mL OXbCD NP 1 group(10μg/mL OXbCD NP incubation followed by PMA stimulation),100μg/mL OXbCD NP 1 group(100μg/mL OXbCD NP incubation followed by PMA stimulation).The intracellular levels of ROS were detected by DCFH-DA staining and flow cytometry,and fluorescence imaging was performed.(2)RAW264.7 cells were randomly divided into 4 groups:blank 2 group(no treatment),control 2 group(IFN-γand LPS stimulation),10μg/mL OXbCD NP 2 group(10μg/mL OXbCD NP incubation followed by IFN-γand LPS stimulation),and 100μg/mL OXbCD NP 2 group(100μg/mL OXbCD NP incubation followed by IFN-γand LPS stimulation).ELISA was used to detect interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)levels.(3)RAW264.7 cells were incubated in OXbCD NP solution at concentrations of 0μg/mL,5μg/mL,10μg/mL,20μg/mL,40μg/mL,80μg/mL,160μg/mL,320μg/mL,640μg/mL and 1280μg/mL for 12 h or 24 h,and cell viability was detected by MTT assay.Results(1)Compared to the control 1 group,ROS levels and fluorescence intensity in the 10μg/mL and 100μg/mL OXbCD NP 1 groups decreased,the difference were statistically significant(P<0.01);(2)Compared to the control 2 group,the levels of IL-1βand TNF-αin the 10μg/mL and 100μg/mL OXbCD NP 2 groups decreased,the difference were statistically significant(P<0.01).(3)The nanoparticles were incubated with the cells for 12 h,compared to OXbCD NP concentration of 0μg/mL(without OXbCD NP),there were statistically significant differences in survival rates when OXbCD NP level reached 640μg/mL and 1280μg/mL(P<0.05).When the incubation time was 24 h,compared to without OXbCD NP,significant differences in cell viability were observed wh

关 键 词：纳米粒 活性氧 细胞氧化应激 

分 类 号：R54[医药卫生—心血管疾病]