miR-93通过激活PI3K/AKT通路调节免疫介导创伤性脑损伤的机制研究

doi:10.19586/j.2095-2341.2025.0027

生物技术进展 ›› 2025, Vol. 15 ›› Issue (4): 711-719.DOI: 10.19586/j.2095-2341.2025.0027

• 研究论文 • 上一篇

miR-93通过激活PI3K/AKT通路调节免疫介导创伤性脑损伤的机制研究

姜晓宇¹(), 陈异², 倪晶晶¹()

^1.苏州大学附属张家港医院急诊科，江苏苏州 215600
^2.新疆医科大学肿瘤医院乳甲外科，乌鲁木齐 830000

收稿日期:2025-03-03 接受日期:2025-04-30 出版日期:2025-07-25 发布日期:2025-09-08
通讯作者: 倪晶晶
作者简介:姜晓宇E-mail: yesterday189@163.com；
基金资助:
新疆维吾尔自治区自然科学基金面上项目(2019D01C269)

Mechanism of miR-93 Regulating Immune-mediated Traumatic Brain Injury Through Activation of PI3K/AKT Pathway

Xiaoyu JIANG¹(), Yi CHEN², Jingjing NI¹()

^1.Department of Emergency，Zhangjiagang Hospital Affiliated to Soochow University，Jiangsu Suzhou 215600，China
^2.Breast and Thyroid Surgery，Cancer Hospital of Xinjiang Medical University，Urumqi 830000，Xinjiang

Received:2025-03-03 Accepted:2025-04-30 Online:2025-07-25 Published:2025-09-08
Contact: Jingjing NI

摘要/Abstract

摘要：

研究旨在探讨miR-93通过PI3K/AKT通路调控巨噬细胞M2极化及神经血管组织修复，以促进创伤性脑损伤（traumatic brain injury，TBI）修复的机制。实验使用雌性C57BL/6小鼠，分为假手术组（Sham组）、模型组（TBI组）、TBI+miR-93抑制剂组和TBI+miR-93激动剂组。qPCR检测M1、M2型巨噬细胞CD80、CD206 mRNA的相对表达水平；酶联免疫反应（enzyme-linked immunosorbent assay, ELISA）检测免疫炎症因子TNF-α、IL-12、TGF-β和IL-13浓度；血管内皮生长因子（vascular endothelial growth factor, VEGF）免疫荧光染色观察TBI后血管组织再生；Western blot检测p-PI3K和p-AKT蛋白的相对表达水平。结果发现，TBI+miR-93激动剂组能促进巨噬细胞M2型极化，抑制TNF-α、IL-12表达，促进TGF-β、IL-13表达，mNSS评分较低且VEGF表达活跃，p-PI3K和p-AKT蛋白表达升高。结果表明，TBI后，miR-93代偿升高，调节巨噬细胞M2型极化，有助于损伤后神经和血管组织修复，这一过程可能是通过激活PI3K/AKT信号通路实现的。

关键词: miR-93, traumatic brain injury, 免疫, 巨噬细胞, PI3K/AKT信号通路

Abstract:

This study aimed to investigate the mechanism by which miR-93 promotes traumatic brain injury （TBI） repair through regulating macrophage M2 polarization and neurovascular tissue regeneration via the PI3K/AKT signaling pathway.Female C57BL/6 mice were randomly assigned to four groups： sham-operated （Sham）， TBI model （TBI）， TBI + miR-93 inhibitor （TBI+inhibitor）， and TBI + miR-93 agonist （TBI+agonist）. Relative mRNA expression levels of M1 macrophage marker CD80 and M2 macrophage marker CD206 were assessed by quantitative real-time PCR （qPCR）. Concentrations of inflammatory cytokines TNF-α， IL-12， TGF-β， and IL-13 were measured using enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence staining for vascular endothelial growth factor （VEGF） was performed to evaluate angiogenesis post-TBI. Western blotting was used to determine the relative protein expression levels of phosphorylated PI3K （p-PI3K） and phosphorylated AKT （p-AKT）.Compared to the TBI group， the TBI+agonist group exhibited enhanced M2 macrophage polarization， reduced expression of pro-inflammatory cytokines TNF-α and IL-12， and increased expression of anti-inflammatory cytokines TGF-β and IL-13. Furthermore， this group showed a lower modified Neurological Severity Score （mNSS）， indicative of improved neurological function， enhanced VEGF expression reflecting active angiogenesis， and elevated levels of p-PI3K and p-AKT proteins.These results suggested that miR-93 expression is compensatorily upregulated following TBI. miR-93 promotes neurovascular repair after injury by modulating macrophage polarization towards the M2 phenotype， likely via activation of the PI3K/AKT signaling pathway.

Key words: miR-93, traumatic brain injury, immunity, macrophages, PI3K/AKT signaling pathway

中图分类号:

姜晓宇, 陈异, 倪晶晶. miR-93通过激活PI3K/AKT通路调节免疫介导创伤性脑损伤的机制研究[J]. 生物技术进展, 2025, 15(4): 711-719.

Xiaoyu JIANG, Yi CHEN, Jingjing NI. Mechanism of miR-93 Regulating Immune-mediated Traumatic Brain Injury Through Activation of PI3K/AKT Pathway[J]. Current Biotechnology, 2025, 15(4): 711-719.

图/表 6

图1 小鼠miR-93和TNF-α的相对表达水平A: RT-qPCR检测TBI后各组miR-93相对表达量；B: TBI后，小鼠miR-93和TNF-α相对表达水平随时间的变化。与Sham组比较，*、***表示差异在P<0.05、P<0.001水平上有统计学意义

Fig. 1 Relative expression levels of miR-93 and TNF-α in mice

图2 巨噬细胞极化表征A：Sham组巨噬细胞向M1型极化；B：TBI组巨噬细胞向M2型极化；C：RT-qPCR检测各组CD80 mRNA（M1型表达）相对表达水平；D：RT-qPCR检测各组CD206 mRNA（M2型表达）相对表达水平。与Sham组比较，*、**、***表示差异在P<0.05、P<0.01、P<0.001水平上有统计学意义。

Fig. 2 Characterization of macrophage polarization

图3 ELISA定量分析结果A：促炎因子TNF-α的相对表达水平；B：促炎因子IL-12的相对表达水平；C：抗炎因子TGF-β的相对表达水平；D；抗炎因子IL-13的相对表达水平。与Sham组比较，*、**、***表示差异在P<0.05、P<0.01、P<0.001水平上有统计学意义。

Fig. 3 Quantitative analysis of ELISA results

图4 术后各组mNSS评分走势图

Fig. 4 Postoperative mNSS score trends in each group

图5 荧光显微镜观察各组血管内皮细胞VEGF的表达情况（×40）注：绿色荧光标记VEGF，蓝色荧光标记细胞核。

Fig. 5 The expression of VEGF in vascular endothelial cells in each group was observed by fluorescence microscopy (×40)

图6 miR-93 对TBI术后PI3K/AKT信号通路的影响A：miR-93处理后各组p-PI3K和p-AKT蛋白表达水平；B~C：p-PI3K和p-AKT的蛋白相对表达量；D~E：p-PI3K和p-AKT的mRNA相对表达量。*、**、***表示与Sham组比较差异在P<0.05、P<0.01、P<0.001水平上有统计学意义

Fig. 6 The effect of miR-93 on the PI3K/AKT signaling pathway after TBI

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miR-93通过激活PI3K/AKT通路调节免疫介导创伤性脑损伤的机制研究

Mechanism of miR-93 Regulating Immune-mediated Traumatic Brain Injury Through Activation of PI3K/AKT Pathway

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