Department of Pediatrics, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Pediatrics, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, China.
Neurol Res. 2023 Dec;45(12):1079-1090. doi: 10.1080/01616412.2023.2257416. Epub 2023 Nov 1.
Hypoxic-ischemic brain damage (HIBD) is among the leading causes of neonatal death worldwide. miR-124-3p can be utilized as a potential diagnostic and prognostic biomarker for perinatal asphyxia and HI encephalopathy in newborns. This study investigated the protective effect and mechanism of miR-124-3p in astrocyte-derived extracellular vesicles (ADEVs) in HIBD.
The neonatal mouse model of HIBD was established. Astrocytes were transfected with the miR-124-3p inhibitor, followed by isolation and identification of ADEVs (ADEVs + inhi miR). HIBD mice were injected with ADEVs or ADEVs + inhi miR through the lateral ventricle, and neurological function was evaluated based on the modified neurological severity score (mNSS). The infarct volume of mice and the morphological modifications of neurons were observed by TTC staining and hematoxylin-eosin staining. The contents of SOD, GSH-Px, CAT, and MDA in the hippocampus were measured. The neuronal apoptosis, the activation of MPO+ neutrophils, NK cells, and CD3+ cells in CA1 region of the hippocampus was determined by means of TUNEL staining and immunofluorescence.
ADEVs alleviated HIBD in neonatal mice. ADEVs could intrinsically protect mice from HIBD by reducing oxidative stress and apoptosis in hippocampal tissue. ADEVs inhibited the positive expression of MPO+ neutrophils, NK cells, and CD3+ cells in HIBD neonatal mice. ADEVs inhibited the hippocampal immune cells by delivering miR-124-3p in neonatal HIBD mice.
ADEVs can inhibit the abnormal activation of immune function in HIBD by delivering miR-124-3p, thereby eliciting a protective effect on brain damage in neonatal mice.
缺氧缺血性脑损伤（HIBD）是全球新生儿死亡的主要原因之一。miR-124-3p 可用作围产期窒息和新生儿 HI 脑病的潜在诊断和预后生物标志物。本研究探讨了 miR-124-3p 在星形胶质细胞衍生的细胞外囊泡（ADEVs）中的保护作用及其在 HIBD 中的作用机制。
建立新生鼠 HIBD 模型。用 miR-124-3p 抑制剂转染星形胶质细胞，分离并鉴定 ADEVs（ADEVs+inhi miR）。通过侧脑室向 HIBD 小鼠注射 ADEVs 或 ADEVs+inhi miR，根据改良神经严重程度评分（mNSS）评估神经功能。通过 TTC 染色和苏木精-伊红染色观察小鼠脑梗死体积和神经元形态学改变。测定海马组织中超氧化物歧化酶（SOD）、谷胱甘肽过氧化物酶（GSH-Px）、过氧化氢酶（CAT）和丙二醛（MDA）的含量。通过 TUNEL 染色和免疫荧光法测定海马 CA1 区神经元凋亡、髓过氧化物酶（MPO）+中性粒细胞、NK 细胞和 CD3+细胞的激活情况。
ADEVs 减轻了新生鼠 HIBD。ADEVs 通过减少海马组织中的氧化应激和细胞凋亡，内在地保护小鼠免受 HIBD 的影响。ADEVs 抑制了 HIBD 新生鼠中 MPO+中性粒细胞、NK 细胞和 CD3+细胞的阳性表达。ADEVs 通过向新生 HIBD 小鼠递送 miR-124-3p 抑制了海马中的免疫细胞。
ADEVs 通过递送 miR-124-3p 抑制 HIBD 中免疫功能的异常激活，从而对新生鼠脑损伤产生保护作用。
