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OACSTPCD
Effects of reactive oxygen species-responsive targeted nanoparticles on cellular oxidative stress and inflammatory cytokines
目的 探讨活性氧(ROS)响应靶向纳米粒(OXbCD 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),10 μg/mL OXbCD NP1组(10 μg/mL OXbCD NP孵育后再加入PMA刺激),100 μg/mL OXbCD NP1 组(100 μg/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),10 μg/mL OXbCD NP 2 组(10 μg/mL OXbCD NP孵育后加入IFN-γ和LPS刺激),100 μg/mL OXbCD NP 2 组(100 μg/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0 μg/mL、5 μg/mL、10 μg/mL、20 μg/mL、40 μg/mL、80 μg/mL、160 μg/mL、320 μg/mL、640 μg/mL、1 280 μg/mL OXbCD 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,10 μg/mL和100 μg/mL 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,10 μg/mL和100 μg/mL 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0 μg/mL(不含OXbCD NP)对比,OXbCD NP水平达到640 μg/mL及1 280 μg/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 320 μg/mL、640 μg/mL及1 280 μg/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 ROS-responsive 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),10 μg/mL OXbCD NP 1 group(10 μg/mL OXbCD NP incubation followed by PMA stimulation),100 μg/mL OXbCD NP 1 group(100 μg/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-γ and LPS stimulation),10 μg/mL OXbCD NP 2 group(10 μg/mL OXbCD NP incubation followed by IFN-γ and LPS stimulation),and 100 μg/mL OXbCD NP 2 group(100 μg/mL OXbCD NP incubation followed by IFN-γ and 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 0 μg/mL,5 μg/mL,10 μg/mL,20 μg/mL,40 μg/mL,80 μg/mL,160 μg/mL,320 μg/mL,640 μg/mL and 1 280 μg/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 10 μg/mL and 100 μg/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-1β and TNF-α in the 10 μg/mL and 100 μg/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 0 μg/mL(without OXbCD NP),there were statistically significant differences in survival rates when OXbCD NP level reached 640 μg/mL and 1 280 μg/mL(P<0.05).When the incubation time was 24 h,compared to without OXbCD NP,significant differences in cell viability were observed when OXbCD NP concentration was 320 μg/mL,640 μg/mL,and 1 280 μg/mL(P<0.05).Conclusion ROS-responsive targeted nanoparticles demonstrate good safety at the cellular level and possess anti-cellular oxidative stress and anti-inflammatory cytokines effects.
曹隆檬;金虎
325000 浙江省温州市中医院心血管科
纳米粒活性氧细胞氧化应激
NanoparticlesReactive oxygen speciesCell oxidative stress
《心脑血管病防治》 2024 (005)
19-22 / 4
温州市科技局课题(Y20210895)
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