内皮祖细胞衍生的条件培养基通过巨噬细胞移动抑制因子激活的 AKT 通路减轻慢性大脑缺血性损伤。

Endothelial progenitor cell-derived conditioned medium mitigates chronic cerebral ischemic injury through macrophage migration inhibitory factor-activated AKT pathway.

机构信息

Division of Neurosurgery, Department of Surgery, National Taiwan University Hospital, No.7, Chung-Shan South Road, Taipei, 100, Taiwan.

Graduate Institute of Clinical Medicine, National Taiwan University College of Medicine, Taipei, Taiwan.

出版信息

Stem Cell Res Ther. 2024 Nov 14;15(1):428. doi: 10.1186/s13287-024-04015-5.

DOI:10.1186/s13287-024-04015-5

PMID:39543689

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11566597/

Abstract

BACKGROUND

Chronic cerebral ischemia (CCI) is a significant health issue characterized by hypoperfusion due to damage or occlusion of the cerebral or carotid arteries. CCI may lead to progressive cognitive impairment that is considered as a prelude to neurodegenerative diseases, including dementia and Alzheimer's disease (AD). Endothelial progenitor cells (EPCs) have been implicated in vascular repair in ischemic cerebrovascular diseases, primarily by differentiating into endothelial cells (ECs) or through paracrine effects. However, the clinical transplantation of stem cell therapies remains limited. In this study, we investigated the effects of EPC-derived conditioned medium (EPC-CM) on the impaired vasculature and neurological function in a rodent model of CCI and the mechanism involved.

METHODS

EPC-CM was analyzed by cytokine array to identify key factors involved in angiogenesis and cellular senescence. The effects and mechanism of the candidate factors in the EPC-CM were validated in vitro using oxygen-glucose deprivation (OGD)-injured ECs and EPCs. The therapeutic effects of EPC-CM and the identified key factor were further examined in a rat model of CCI, which was induced by bilateral internal carotid artery ligation (BICAL). EPC-CM was administered via intracisternal injection one week post BICAL. The cerebral microvasculature and neurobehavior of the rats were examined three weeks after BICAL.

RESULTS

Macrophage migration inhibitory factor (MIF) was identified as a key factor in the EPC-CM. Recombinant MIF protein promoted angiogenesis and prevented senescence in the injured EPCs and ECs. The effect was similar to that of the EPC-CM. These therapeutic effects were diminished when the EPC-CM was co-treated with MIF-specific antibody (Ab). Additionally, the vascular, motor, and cognitive improvements observed in the BICAL rats treated with EPC-CM were abolished by co-treated with MIF Ab. Furthermore, we found MIF promoted angiogenesis and anti-senescence via activating the AKT pathway. Inhibition of the AKT pathway diminished the protective effects of MIF in the in vitro study.

CONCLUSIONS

We demonstrated that EPC-CM protected the brain from chronic ischemic injury and promoted functional recovery through MIF-mediated AKT pathway. These findings suggest EPC-CM holds potential as a novel cell-free therapeutic approach for treating CCI through the actions of MIF.

摘要

背景

慢性脑缺血（CCI）是一种严重的健康问题，其特征是由于大脑或颈动脉的损伤或闭塞导致灌注不足。CCI 可能导致进行性认知障碍，被认为是神经退行性疾病（包括痴呆和阿尔茨海默病（AD））的前奏。内皮祖细胞（EPCs）已被认为参与缺血性脑血管疾病中的血管修复，主要通过分化为内皮细胞（ECs）或通过旁分泌作用。然而，干细胞治疗的临床移植仍然受到限制。在这项研究中，我们研究了 EPC 衍生的条件培养基（EPC-CM）对 CCI 啮齿动物模型中受损血管和神经功能的影响及其涉及的机制。

方法

通过细胞因子阵列分析 EPC-CM，以鉴定参与血管生成和细胞衰老的关键因素。使用氧葡萄糖剥夺（OGD）损伤的 ECs 和 EPCs 在体外验证 EPC-CM 中候选因子的作用和机制。通过双侧颈内动脉结扎（BICAL）诱导 CCI 大鼠模型进一步研究 EPC-CM 和鉴定的关键因子的治疗效果。EPC-CM 通过颅内注射给药，在 BICAL 后一周进行。BICAL 后三周检查大鼠的脑微血管和神经行为。

结果

巨噬细胞移动抑制因子（MIF）被鉴定为 EPC-CM 中的关键因素。重组 MIF 蛋白促进了受损 EPCs 和 ECs 的血管生成并防止了衰老。这种作用与 EPC-CM 相似。当 EPC-CM 与 MIF 特异性抗体（Ab）共同处理时，这种治疗效果会减弱。此外，用 MIF Ab 共同处理可消除 BICAL 大鼠中用 EPC-CM 观察到的血管、运动和认知改善。此外，我们发现 MIF 通过激活 AKT 途径促进血管生成和抗衰老。在体外研究中，AKT 途径的抑制减弱了 MIF 的保护作用。