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通过强制表达miR-124和生长因子处理上调神经祖细胞和神经标志物

Upregulation of Neuroprogenitor and Neural Markers via Enforced miR-124 and Growth Factor Treatment.

作者信息

Mehri Ghahfarrokhi Ameneh, Jami Mohammad Saeid, Hashemzadeh Chaleshtori Morteza

机构信息

Department of Molecular Medicine, School of Advanced Technologies, Shahrekord University of Medical Sciences, Shahrekord, Iran.

Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.

出版信息

Int J Mol Cell Med. 2020 Winter;9(1):62-70. doi: 10.22088/IJMCM.BUMS.9.1.62.

Abstract

Previous studies have shown that miR-124 plays an important role in the development of auditory neurons, which are degenerated in the sensorineural hearing loss. However, whether the combined use of miR-124 and growth factors can increase the expression of neural related markers in human dental pulp stem cells has been remained unknown so far. In this study, human dental pulp stem cells were transfected with miR-124 following treatment with brain-derived neurotrophic factor or epidermal growth factor/basic fibroblast growth factor. The expression of some neural related markers (nestin, , β-tubulin III, , and peripherin) was analyzed in two groups by qRT-PCR or immunofluorescence. Cellular treatment resulted in morphological changes including neurosphere-like colonies formation. Nestin and were up-regulated, and and peripherin were down-regulated in dental pulp stem cells transfected by miR-124 following treatment with brain-derived neurotrophic factor. Replacement of brain-derived neurotrophic factor with epidermal growth factor/ basic fibroblast growth factor resulted in the up-regulation of , , peripherin, and β-tubulin III and down-regulation of . The expression of SOX2 and nestin was also confirmed by immunofluorescence. The combination of miR-124 and growth factors would provide a promising starting point for upregulating the neural progenitor markers in human dental pulp stem cells.

摘要

先前的研究表明,miR-124在听觉神经元的发育中起重要作用,而听觉神经元在感音神经性听力损失中会退化。然而,miR-124与生长因子联合使用是否能增加人牙髓干细胞中神经相关标志物的表达,迄今为止仍不清楚。在本研究中,在用脑源性神经营养因子或表皮生长因子/碱性成纤维细胞生长因子处理后,将miR-124转染到人牙髓干细胞中。通过qRT-PCR或免疫荧光分析两组中一些神经相关标志物(巢蛋白、β-微管蛋白III和外周蛋白)的表达。细胞处理导致形态学改变,包括形成神经球样集落。在用脑源性神经营养因子处理后,转染miR-124的牙髓干细胞中巢蛋白和 上调,而 和外周蛋白下调。用表皮生长因子/碱性成纤维细胞生长因子替代脑源性神经营养因子导致 、 、外周蛋白和β-微管蛋白III上调,而 下调。免疫荧光也证实了SOX2和巢蛋白的表达。miR-124与生长因子的联合使用将为上调人牙髓干细胞中的神经祖细胞标志物提供一个有前景的起点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bd26/7422846/8559ea18281e/ijmcm-9-62-g001.jpg

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