Department of Pediatrics, University of Florida, Gainesville, Florida, USA.
Department of Biomolecular Sciences, University of Urbino Carlo Bo, Urbino, Italy.
J Pineal Res. 2022 Sep;73(2):e12818. doi: 10.1111/jpi.12818. Epub 2022 Jul 22.
Neonatal encephalopathy (NE) is a pathological condition affecting long-term neurodevelopmental outcomes. Hypothermia is the only therapeutic option, but does not always improve outcomes; hence, researchers continue to hunt for pharmaceutical compounds. Melatonin treatment has benefitted neonates with hypoxic-ischemic (HI) brain injury. However, unlike animal models that enable the study of the brain and the pathophysiologic cascade, only blood is available from human subjects. Therefore, due to the unavailability of neonatal brain tissue, assumptions about the pathophysiology in pathways and cascades are made in human subjects with NE. We analyzed animal and human specimens to improve our understanding of the pathophysiology in human neonates. A neonate with NE who underwent hypothermia and enrolled in a melatonin pharmacokinetic study was compared to HI rats treated/untreated with melatonin. MicroRNA (miRNA) analyses provided profiles of the neonate's plasma, rat plasma, and rat brain cortexes. We compared these profiles through a bioinformatics tool, identifying Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways common to HI brain injury and melatonin treatment. After evaluating the resulting pathways and the literature, to validate the method, the key proteins expressed in HI brain injury were investigated using cerebral cortexes. The upregulated miRNAs in human neonate and rat plasma helped identify two KEGG pathways, glioma and long-term potentiation, common to HI injury and melatonin treatment. A unified neonatal cerebral melatonin-sensitive HI pathway was designed and validated by assessing the expression of protein kinase Cα (PKCα), phospho (p)-Akt, and p-ERK proteins in rat brain cortexes. PKCα increased in HI-injured rats and further increased with melatonin. p-Akt and p-ERK returned phosphorylated to their basal level with melatonin treatment after HI injury. The bioinformatics analyses validated by key protein expression identified pathways common to HI brain injury and melatonin treatment. This approach helped complete pathways in neonates with NE by integrating information from animal models of HI brain injury.
新生儿脑病 (NE) 是一种影响长期神经发育结局的病理状况。 低温是唯一的治疗选择，但并不总是能改善结局；因此，研究人员继续寻找药物化合物。 褪黑素治疗已使患有缺氧缺血性 (HI) 脑损伤的新生儿受益。 然而，与允许研究大脑和病理生理级联的动物模型不同，人类受试者只有血液可用。 因此，由于无法获得新生儿脑组织，因此在患有 NE 的人类受试者中对途径和级联中的病理生理学做出了假设。 我们分析了动物和人类标本，以增进对人类新生儿病理生理学的理解。 将接受低温治疗并参加褪黑素药代动力学研究的 NE 新生儿与接受/未接受褪黑素治疗的 HI 大鼠进行比较。 微 RNA (miRNA) 分析提供了新生儿血浆、大鼠血浆和大鼠大脑皮质的图谱。 我们通过生物信息学工具比较了这些图谱，确定了与 HI 脑损伤和褪黑素治疗相关的京都基因与基因组百科全书 (KEGG) 途径。 在评估了产生的途径和文献后，为了验证该方法，使用大脑皮质研究了 HI 脑损伤中表达的关键蛋白。 人类新生儿和大鼠血浆中上调的 miRNA 有助于确定两个与 HI 损伤和褪黑素治疗相关的 KEGG 途径，即神经胶质瘤和长时程增强。 通过评估大鼠大脑皮质中蛋白激酶 Cα (PKCα)、磷酸化 (p)-Akt 和 p-ERK 蛋白的表达，设计并验证了针对 HI 损伤的统一新生儿大脑褪黑素敏感 HI 途径。 在 HI 损伤的大鼠中 PKCα 增加，并且在用褪黑素进一步增加。 HI 损伤后，用褪黑素处理可使 p-Akt 和 p-ERK 返回到其基础水平。 通过关键蛋白表达验证的生物信息学分析确定了与 HI 脑损伤和褪黑素治疗相关的途径。 通过整合 HI 脑损伤动物模型的信息，这种方法有助于通过整合 HI 脑损伤动物模型的信息来完成 NE 新生儿的途径。
