Deng Li, Gao Xiaoqing, Fan Guangbi, Yang Chaoxian
Department of Neurobiology, Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan, China.
Department of Neurobiology, Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan, China.
J Stroke Cerebrovasc Dis. 2019 Sep;28(9):2555-2562. doi: 10.1016/j.jstrokecerebrovasdis.2019.06.002. Epub 2019 Jun 24.
The present study aimed to investigate the effects of Mesenchymal stem cells/glial cell line derived neurotrophic factor (MSCs/GDNF) transplantation on nerve reconstruction in rats with intracerebral hemorrhage.
GDNF transduction to MSCs was using adenovirus vector pAdEasy-1-pAdTrack-CMV prepared. Intracerebral hemorrhage (ICH) was induced by injection of collagenase and heparin into the caudate putamen. At the third day after a collagenase-induced ICH, adult male SD rats were randomly divided into saline group, MSCs group and MSCs/GDNF group. Immunofluorescence and RT-PCR were performed to detect the differentiation of MSCs or MSCs with an adenovirus vector encoding GDNF gene in vivo and in vitro.
After 6 hours of induction, both MSCs and MSCs/GDNF expressed neuro or glial specific markers and synaptic-associated proteins (SYN, GAP-43, PSD-95); additionally, they secreted bioactive compounds (BDNF, NGF-β). MSCs/GDNF transplantation, compared to MSCs and saline solution injection, significantly improved neurological functions after ICH. The grafted MSCs or MSCs/GDNF survived in the striatum after 2 weeks of transplantation and expressed the neural cell-specific biomarkers NSE, MAP2, and GFAP.
These findings demonstrate that MSCs/GDNF transplantation contributes to improved neurological function in experimental ICH rats. The mechanisms are possibly due to neuronal replacement and enhanced neurotrophic factor secretion.
本研究旨在探讨间充质干细胞/胶质细胞源性神经营养因子(MSCs/GDNF)移植对脑出血大鼠神经重建的影响。
采用腺病毒载体pAdEasy-1-pAdTrack-CMV将GDNF转导至MSCs。通过向尾状核壳核注射胶原酶和肝素诱导脑出血。在胶原酶诱导脑出血后的第3天,将成年雄性SD大鼠随机分为生理盐水组、MSCs组和MSCs/GDNF组。采用免疫荧光和RT-PCR检测MSCs或携带GDNF基因腺病毒载体的MSCs在体内和体外的分化情况。
诱导6小时后,MSCs和MSCs/GDNF均表达神经或胶质特异性标志物以及突触相关蛋白(SYN、GAP-43、PSD-95);此外,它们还分泌生物活性化合物(BDNF、NGF-β)。与MSCs和注射生理盐水相比,MSCs/GDNF移植显著改善了脑出血后的神经功能。移植2周后,移植的MSCs或MSCs/GDNF在纹状体中存活并表达神经细胞特异性生物标志物NSE、MAP2和GFAP。
这些发现表明,MSCs/GDNF移植有助于改善实验性脑出血大鼠的神经功能。其机制可能是神经元替代和神经营养因子分泌增加。