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

LEI Hangke; HUANG Dingde

doi:10.16016/j.2097-0927.202411116

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

Military Medicine | 更新时间：2025-03-28

- Preparation of reactive oxygen species scavenging Ce6-Luminol encapsulated liposomes and their protective effect against radiation pneumonitis in mice
- Journal of Army Medical University Vol. 47, Issue 6, Pages: 489-497(2025)
- 作者机构：
  
  陆军军医大学（第三军医大学）第一附属医院核医学科，重庆
- 作者简介：
  
  HUANG Dingde， E-mail： huangdingde@126.com
- 基金信息：
  
  the General Project of the Natural Science Foundation of Chongqing(CSTB2022NSCQ-MSX0150)
- DOI：10.16016/j.2097-0927.202411116
  CLC： R563.1;R818.84;R945
- Received：26 November 2024，
  
  Revised：14 December 2024，
  
  Published：30 March 2025
- 稿件说明：
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雷航科，黄定德.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［J］.陆军军医大学学报，2025，47（06）：489-497.DOI：,10.16016/j.2097-0927.202411116.

LEI H K，HUANG D D.Preparation of reactive oxygen species scavenging Ce6-Luminol encapsulated liposomes and their protective effect on radiation pneumonitis in mice［J］.J Army Med Univ，2025，47（06）：489-497.DOI：,10.16016/j.2097-0927.202411116.
雷航科，黄定德.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［J］.陆军军医大学学报，2025，47（06）：489-497.DOI：,10.16016/j.2097-0927.202411116. DOI：

LEI H K，HUANG D D.Preparation of reactive oxygen species scavenging Ce6-Luminol encapsulated liposomes and their protective effect on radiation pneumonitis in mice［J］.J Army Med Univ，2025，47（06）：489-497.DOI：,10.16016/j.2097-0927.202411116. DOI：

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C

e6-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

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（

=351.741，

0.05）。CLL干预可显著降低放射性肺炎小鼠肺组织活性氧H

（

=15.183，

0.05）、炎症因子TNF-α （

=5.150，

0.05）和IL-1β（

=10.036，

0.05）、炎症细胞趋化因子MCP-1（

=3.777，

0.05）和KC（

=3.755，

0.05）的含量，以及支气管肺泡灌洗液中性粒细胞比例（

=17.751，

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。

Abstract

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

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

and 5 post-irradiation. Then on day 7 post-irradiation

the contents of H

inflammatory cytokines TNF-α and 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 pneumonitis mice than in normal mice (

=351.741

0.05). CLL intervention significantly reduced the lung contents of H

(

=15.183

0.05)

TNF-α (

=5.150

0.05)

IL-1β (

=10.036

0.05)

MCP-1 (

=3.777

0.05) and KC (

=3.755

0.05)

and the proportion of neutrophils in BALF (

=17.751

0.05) from the radiation pneumonitis mice. Conclusion‍　‍ROS scavenging CLL liposomes can alleviate radiation pneumoni

tis in mice

which may be due to the protective role against radiation pneumonitis.

关键词

Keywords

references

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