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Abstract：
This study aimed to decipher the effect of glycoprotein nonmetastatic melanoma protein B (GPNMB) on neonatal hypoxic-ischemic encephalopathy (NHIE) and its potential molecular mechanism. The hypoxic-ischemic (HI) model was established in 7-day-old rats, and then, Zea-Longa scores and Nissl staining were performed to measure brain damage post-HI. In addition, gene sequencing was used to detect the differential expression genes (DEGs), and then, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes databases were used to determine the function of DEGs. Furthermore, an oxygen-glucose deprivation (OGD) model was developed in SY5Y cells and human fetal neurons, and then, the level of GPNMB was verified by quantitative real-time polymerase chain reaction. In addition, methyl thiazolyl tetrazolium and cell counting kit-8 assays were applied after GPNMB interference. Finally, the alternative splicing of GPNMB expression was analyzed using Splice Grapher software. The results indicated that HI induced marked neurological impairment and neuron injury in rats. Also, GPNMB was the most obviously upregulated gene in DEGs. Additionally, GPNMB was upregulated significantly in SY5Y and fetal neurons after OGD, and GPNMB-si promoted an increase in cell viability and number. Moreover, we found that the GPNMB alternative splicing type was the Alternative 3\' splice site, with the alternative splicing site in 143382985:143404102. Herein, GPNMB promotes a crucial regulatory mechanism with alternative splicing for neuronal survival after NHIE.
摘要：
本研究旨在破译糖蛋白非转移性黑色素瘤蛋白B(GPNMB)对新生儿缺氧缺血性脑病(NHIE)的影响及其可能的分子机制。建立7日龄大鼠缺氧缺血(HI)模型,然后,进行Zea-Longa评分和Nissl染色以测量HI后的脑损伤。此外,基因测序用于检测差异表达基因(DEGs)，然后,基因本体论和京都百科全书的基因和基因组数据库用于确定DEG的功能。此外，在SY5Y细胞和人胎儿神经元中建立了氧葡萄糖剥夺（OGD）模型，然后,通过定量实时聚合酶链反应验证GPNMB水平.此外,GPNMB干扰后应用甲基噻唑基四唑和细胞计数试剂盒-8测定。最后,使用SpliceGrapher软件分析GPNMB表达的选择性剪接。结果表明，HI可引起大鼠明显的神经功能缺损和神经元损伤。此外，GPNMB是DEGs中最明显的上调基因。此外,OGD后，SY5Y和胎儿神经元中GPNMB显著上调，和GPNMB-si促进细胞活力和数量的增加。此外，我们发现GPNMB选择性剪接类型是选择性3\'剪接位点，在143382985:143404102中具有选择性剪接位点。在这里，GPNMB通过选择性剪接促进NHIE后神经元存活的关键调节机制。
