上海交通大学学报（医学版）

上海交通大学学报（医学版） >

2025 , Vol. 45 >Issue 9: 1161 - 1170

DOI: https://doi.org/10.3969/j.issn.1674-8115.2025.09.008

论著 · 基础研究

负载白细胞介素-4的水凝胶微球调控卒中后免疫微环境对神经功能修复的影响

  • 徐彤彤 ,
  • 阮慧瞳
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  • 1.上海交通大学医学院附属瑞金医院骨科，上海市伤骨科研究所，上海 200025
    2.上海交通大学生物医学工程学院，上海 200030
    3.上海交通大学医学院医学技术学院，上海 200025
阮慧瞳，副研究员，博士；电子信箱：ruanhuitong@163.com/ruanhuitong@sjtu.edu.cn。

收稿日期: 2025-05-05

  录用日期: 2025-08-16

  网络出版日期: 2025-09-30

基金资助

国家自然科学基金(82372120);上海交通大学医学院“双百人”项目(2024-0701)

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Effects of hydrogel microspheres loaded with interleukin-4 on neural functional recovery by modulating the immune microenvironment after stroke

  • XU Tongtong ,
  • RUAN Huitong
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  • 1.Shanghai Institute of Traumatology and Orthopaedics, Department of Orthopaedics, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
    2.School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China
    3.College of Health Science and Technology, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
RUAN Huitong, E-mail: ruanhuitong@163.com/ruanhuitong@sjtu.edu.cn.

Received date: 2025-05-05

  Accepted date: 2025-08-16

  Online published: 2025-09-30

Supported by

National Natural Science Foundation of China(82372120);“Two-hundred Talents” Program of Shanghai Jiao Tong University School of Medicine(2024-0701)

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摘要

目的·探讨负载免疫调控因子白细胞介素-4（interleukin-4，IL-4）的水凝胶微球（microsphere，MP）对卒中损伤后再生与修复的影响。方法·通过乳化法生成负载IL-4的纳米颗粒，并联合气流控法构建的聚乙烯醇（polyvinyl alcohol，PVA）水凝胶微球制得负载IL-4的PVA水凝胶微球（MP@IL-4）。使用光学显微镜对MP@IL-4的形态进行表征。体外培养BV2小胶质细胞，经MP@IL-4处理后，分别采用CCK-8法、活/死细胞染色法对其细胞活力、细胞存活进行检测。经脂多糖继续处理BV2小胶质细胞后，采用免疫荧光染色对其促炎标志物诱导型一氧化氮合酶（inducible nitric oxide synthase，iNOS）和抑炎标志物精氨酸酶1（arginase 1，Arg-1）的表达水平进行分析。同时，采用大脑中动脉栓塞法构建小鼠卒中模型，随机分为假手术组（Sham组）、缺血性卒中（ischemic stroke，IS）组和微球治疗组（IS-MP@IL-4组）。通过神经行为学评分、悬空旋转实验和悬丝实验评估3组小鼠的神经行为学功能，焦油紫染色检测3组小鼠的脑萎缩体积，免疫荧光染色表征3组小鼠小胶质细胞促炎因子CD86和抑炎因子CD206的表达水平。结果·光学显微镜的观察结果显示成功制备了粒径为200 μm的MP@IL-4，且不影响BV2小胶质细胞的细胞活力和存活。体外和体内实验均证实，MP@IL-4可下调小胶质细胞的促炎标志物的表达，上调抑炎标志物的表达。此外，与IS组相比，IS-MP@IL-4组小鼠的神经行为学功能有较大改善（P<0.05），脑萎缩体积也有所减小（P<0.001）。结论·MP@IL-4可通过改善免疫微环境发挥对卒中后神经功能损伤的治疗作用。

关键词： 水凝胶微球; 药物递送; 缺血性卒中; 抑炎作用; 神经修复

本文引用格式

徐彤彤 , 阮慧瞳 . 负载白细胞介素-4的水凝胶微球调控卒中后免疫微环境对神经功能修复的影响[J]. 上海交通大学学报（医学版）, 2025 , 45(9) : 1161 -1170 . DOI: 10.3969/j.issn.1674-8115.2025.09.008

Abstract

Objective ·To explore the effects of hydrogel microsphere (MP) loaded with the immunoregulatory factor interleukin-4 (IL-4) on the regeneration and repair after stroke injury. Methods ·IL-4-loaded nanoparticles were prepared using an emulsification method and subsequently incorporated into polyvinyl alcohol (PVA) hydrogel microspheres via an airflow technique to obtain IL-4-loaded PVA hydrogel microspheres (MP@IL-4). The morphology of MP@IL-4 was characterized by optical microscopy. BV2 microglia were cultured in vitro and treated with MP@IL-4. Cell viability and survival were detected by the CCK-8 assay and live/dead cell staining, respectively. Subsequently, BV2 microglia were further treated with lipopolysaccharide (LPS), and the expression levels of the pro-inflammatory marker inducible nitric oxide synthase (iNOS) and the anti-inflammatory marker arginase 1 (Arg-1) were detected by immunofluorescence staining. Meanwhile, a mouse stroke model was constructed using the middle cerebral artery occlusion method, and the mice were randomly divided into the sham operation group (Sham group), the ischemic stroke (IS) group and the microsphere treatment group (IS-MP@IL-4 group). The neurobehavioral functions of mice in the three groups were detected by the modified Neurological Severity Score, elevated body swing test, and hanging wire test. The brain atrophy volume was detected by cresyl violet staining. Immunofluorescence staining was used to assess the expression levels of the pro-inflammatory factor CD86 and the anti-inflammatory factor CD206 in microglia. Results ·Optical microscopy confirmed the successful fabrication of MP@IL-4 with a particle size of 200 μm, which did not affect the cell viability and survival of BV2 microglia. Both in vitro and in vivo experiments demonstrated that MP@IL-4 downregulated the expression of pro-inflammatory markers and upregulated the expression of anti-inflammatory markers in microglia. Moreover, compared with the IS group, the neurobehavioral function of mice in the IS-MP@IL-4 group was significantly improved (P<0.05), and the brain atrophy volume was reduced (P<0.001). Conclusion ·MP@IL-4 can exert a therapeutic effect on post-stroke neurofunctional injury by improving the immune microenvironment.

Key words： hydrogel microsphere; drug delivery; ischemic stroke (IS); anti-inflammatory effect; neural repair

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