Zeng Wen, Wang Yu, Xi Yufeng, Wei Guoqing, Ju Rong
Department of Neonatology, Chengdu Women's and Children's Central Hospital, School of Medicine, University of Electronic Science and Technology of China, Chengdu, 611731, Sichuan, China.
Transl Neurosci. 2021 Dec 17;12(1):561-572. doi: 10.1515/tnsci-2020-0204. eCollection 2021 Jan 1.
Hypoxic-ischemic brain damage (HIBD) is a major cause of brain injury in neonates. Bone marrow mesenchymal stem cells (BMSCs) show therapeutic potential for HIBD, and genetic modification may enhance their neuroprotective effects. The goal of this study was to investigate the neuroprotective effects of hepatocyte growth factor (HGF)-overexpressing BMSCs (BMSCs-HGF) against HIBD and their underlying mechanisms.
BMSCs were transfected with HGF using adenoviral vectors. HIBD models were established and then BMSCs were transplanted into the brains of HIBD rats via intraventricular injection. 2,3,5-Triphenyltetrazolium chloride (TTC) staining was used to measure cerebral infarction volumes. , primary cultured cortical neurons were co-cultured with BMSCs in a Transwell plate system. Oxygen-glucose deprivation (OGD) was applied to imitate hypoxic-ischemic insult, and PD98059 was added to the culture medium to block the phosphorylation of extracellular signal-regulated kinase (ERK). Cell apoptosis was determined using TUNEL staining. The expression of HGF was measured by immunofluorescence, real-time quantitative PCR (RT-qPCR), and western blots. The expression of phosphorylated ERK (p-ERK) and B-cell lymphoma-2 (Bcl-2) was measured by western blots.
HGF-gene transfection promoted BMSC proliferation. Moreover, BMSCs-HGF decreased HIBD-induced cerebral infarction volumes and enhanced the protective effects of the BMSCs against HIBD. BMSCs-HGF also increased expression of HGF, p-ERK, and Bcl-2 in brain tissues. , BMSC-HGF protected neurons against OGD-induced apoptosis. Inhibition of ERK phosphorylation abolished the neuroprotective effect of BMSCs-HGF against OGD.
BMSCs-HGF is a potential treatment for HIBD and that the ERK/Bcl-2 pathway is involved in the underlying neuroprotective mechanism.
缺氧缺血性脑损伤（HIBD）是新生儿脑损伤的主要原因。骨髓间充质干细胞（BMSCs）对HIBD显示出治疗潜力，基因修饰可能增强其神经保护作用。本研究的目的是探讨过表达肝细胞生长因子（HGF）的BMSCs（BMSCs-HGF）对HIBD的神经保护作用及其潜在机制。
使用腺病毒载体将HGF转染至BMSCs。建立HIBD模型，然后通过脑室内注射将BMSCs移植到HIBD大鼠脑内。采用2,3,5-氯化三苯基四氮唑（TTC）染色测量脑梗死体积。原代培养的皮质神经元在Transwell板系统中与BMSCs共培养。应用氧糖剥夺（OGD）模拟缺氧缺血性损伤，并在培养基中加入PD98059以阻断细胞外信号调节激酶（ERK）的磷酸化。使用TUNEL染色测定细胞凋亡。通过免疫荧光、实时定量PCR（RT-qPCR）和蛋白质印迹法测量HGF的表达。通过蛋白质印迹法测量磷酸化ERK（p-ERK）和B细胞淋巴瘤-2（Bcl-2）的表达。
HGF基因转染促进了BMSCs增殖。此外，BMSCs-HGF减少了HIBD诱导的脑梗死体积，并增强了BMSCs对HIBD的保护作用。BMSCs-HGF还增加了脑组织中HGF、p-ERK和Bcl-2的表达。BMSC-HGF保护神经元免受OGD诱导的凋亡。抑制ERK磷酸化消除了BMSCs-HGF对OGD的神经保护作用。
BMSCs-HGF是HIBD的一种潜在治疗方法，ERK/Bcl-2通路参与了潜在的神经保护机制。
