高表达miR-135a的骨髓间充质干细胞来源的外泌体对细菌性脑膜炎新生大鼠神经损伤及FOXO1表达的影响OA
Effect of BMSC-derived exosomes with high miR-135a on neural injury and FOXO1 expression of neonatal rats with bacterial meningitis
目的:探讨高表达 miR-135a 的骨髓间充质干细胞来源外泌体(BMSC Exo-miR-135a)对细菌性脑膜炎(BM)新生大鼠神经损伤及 FOXO1 表达的影响.方法:分离大鼠 BMSC,用脂质体 3000 将 miR-135 mimic 或 miR-NC 转染48 h 后,密度梯度离心法分离外泌体,得 BMSC Exo-miR-135a 和 Exo-miR-NC.60 只新生大鼠随机分为对照、BM、Exo-miR-NC 和 Exo-miR-135a 组,每组15 只.BM、Exo-miR-NC 和 Exo-miR-135a 组小脑延髓池内注射 B 族链球菌(GBS),6 h 后分别向侧脑室内注射生理盐水、Exo-miR-NC 和 Exo-miR-135a.GBS 注射24 h 后,Loeffler 评分法进行神经行为学评估,收集脑脊液进行GBS 菌落、白细胞计数,ELISA 法检测TNF-α、IL-1β 和IL-6 含量(n=15);EB 实验检测血脑屏障通透性(n=5),HE、Nissl 和 TUNEL 染色评估脑组织病理学改变(n=5);RT-qPCR 检测脑组织中 miR-135a 和 FOXO1 mRNA 表达,Western blot 法检测 FOXO1 蛋白的表达(n=5).结果:与对照组比较,BM 组大鼠 Loeffler 评分降低;脑脊液中 GBS 菌落计数、白细胞计数和炎症因子含量,EB 渗漏量,脑含水量增加,脑组织Nissl 小体数量和 miR-135a 表达降低,HE 评分、细胞凋亡率及 FOXO1 表达增加(P＜0.05).与 BM 组相比,Exo-miR-NC 和 Exo-miR-135a 处理可逆转上述指标变化(P＜0.05),Exo-miR-135a 组变化更显著(P＜0.05).结论:BM-SC Exo-miR-135a 可能通过靶向 FOXO1 减轻炎症反应和细胞凋亡,对 BM 新生大鼠发挥神经保护作用Nissl 小体数量和 miR-135a 表达降低,HE 评分、细胞凋亡率及 FOXO1 表达增加(P＜0.05).与 BM 组相比,Exo-miR-NC 和 Exo-miR-135a 处理可逆转上述指标变化(P＜0.05),Exo-miR-135a 组变化更显著(P＜0.05).结论:BM-SC Exo-miR-135a 可能通过靶向 FOXO1 减轻炎症反应和细胞凋亡,对 BM 新生大鼠发挥神经保护作用.
Aim:To investigate the effect of bone marrow mesenchymal stem cell-derived exosomes with high miR-135a expression(BMSC Exo-miR-135a)on neural injury and FOXO1 expression of neonatal rats with bacterial meningitis(BM).Methods:Rat BMSC were isolated and transfected with miR-135 mimic or miR-NC using liposome 3000 for 48 hours,exo-somes were isolated by density gradient centrifugation to obtain BMSC Exo-miR-135a and Exo-miR-NC.Sixty neonatal rats were randomly divided into a control group,BM group,Exo-miR-NC group,and Exo-miR-135a group,with 15 rats in each group.The cerebellomedullary pool of the BM group,Exo-miR-NC group,and Exo-miR-135a group was injected with group B streptococcus(GBS),6 hours later,physiological saline,Exo-miR-NC,and Exo-miR-135a were injected into the lateral ventricles,respectively.24 hours after GBS injection,neurobehavioral assessments were performed using the Loeffler scoring system,cerebrospinal fluid was collected to determine GBS colony count and white blood cell(WBC)count,and the levels of TNF-α,IL-1β,and IL-6 were measured by ELISA(n=15).Blood-brain barrier permeability was evaluated using EB as-say(n=5),and pathological changes in brain tissue were assessed by HE,Nissl,and TUNEL staining(n=5).RT-qPCR was used to detect the expression levels of miR-135a and FOXO1 mRNA in brain tissue,and Western blot was employed to detect FOXO1 protein expression(n=5).Results:Compared with the control group,the BM group rats exhibited reduced Loeffler score;increased GBS colony count,WBC count,inflammatory cytokine levels in cerebrospinal fluid,increased EB leakage volume,brain water content;fewer Nissl bodies and decreased miR-135a expression in brain tissue;increased HE staining score,apoptosis rate,and FOXO1 expression in brain tissue(P＜0.05).Treatment with Exo-miR-NC and Exo-miR-135a reversed these changes compared with the BM group(P＜0.05),with Exo-miR-135a treatment demonstrating more significant effects(P＜0.05).Conclusion:BMSC-Exo-miR-135a may exert neuroprotective effects on neonatal BM rats by targeting FOXO1 to mitigate inflammation and apoptosis.
李月云;王艳蕊;付艳
新乡市中心医院儿科 河南 新乡 453000新乡市中心医院儿科 河南 新乡 453000新乡市中心医院儿科 河南 新乡 453000
医药卫生
骨髓间充质干细胞外泌体miR-135aFOXO1细菌性脑膜炎神经损伤
bone marrow mesenchymal stem cellsexosomemiR-135aFOXO1bacterial meningitisneuronal damage
《郑州大学学报（医学版）》 2026 (3)
48-53,6
河南省医学科技攻关计划联合共建项目(LHGJ20230892)
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