新生儿缺氧缺血性脑病中的生物节律紊乱及其机制研究

发布时间：2018-04-27 04:30

本文选题：睡眠 + 缺氧缺血脑病　；参考：《苏州大学》2016年博士论文

【摘要】：新生儿缺氧缺血性脑病(HIE)是新生儿窒息常见的严重并发症。但是,不同程度HIE患儿存在的昼夜节律问题仍未受到重视,对其机制更知之甚少。因此,进一步研究不同程度HIE患儿的睡眠障碍问题,以及可能的参与机制,对于完善治疗方案和提高患儿的生存质量,减轻家庭负担,具有重要的临床价值和意义。miRNAs作为重要的基因表达调节子与多种疾病的发生、发展有着密切的联系,但miRNAs在HIE中的作用及其机制却尚未见报道。本研究通过对不同程度的HIE病人的睡眠状况的随访研究及对动物模型的松果体组织的系统研究,探讨筛选出的miR-182在缺氧缺血后松果体功能损伤导致的昼夜节律异常中的作用机制,以期为HIE患儿的睡眠障碍的生物治疗提供新的靶点。第一部分轻中度HIE对儿童睡眠的影响目的：本部分研究旨在阐明非重度围产期缺氧缺血损伤后患儿的睡眠紊乱的特点。方法：随访2010年3月至2013年8月间转院至我苏州大学附属儿童医院住院,并符合入选标准的有缺氧缺血脑病史的新生儿128名,同时,以同期年龄和性别相匹配的88名正常足月新生儿作为对照,收集患者的临床资料,并进行问卷调查评估睡眠情况。用Wilcoxon符号等级检验进行比较,探索围产期不同程度HIE患儿的睡眠紊乱问题。结果：通过对不同程度HIE患儿睡眠治疗的调查,发现：轻中度HIE患儿不同程度地表现出明显的睡眠的发动和维持、睡眠相关的呼吸问题以及昼夜节律问题。1、与轻度HIE患儿相比,中度HIE患儿将来发生睡眠呼吸问题的风险明显增高；2、轻度HIE患儿比中度HIE患儿发生昼夜节律紊乱的几率较高：结论：本研究结果提示轻中度HIE患儿不同程度地表现出明显的睡眠发动和维持、睡眠相关的呼吸问题以及昼夜节律问题,由此为HIE患儿通过睡眠障碍的靶向治疗改善远期预后的设想提供了的临床依据。第二部分新生大鼠HIBD后miR-182在调节松果体CLOCK表达中的作用目的：通过建立体内和体外新生大鼠缺氧缺血脑损伤(HIBD)模型,分析miR-182在松果体功能损伤导致的昼夜节律紊乱中的作用机制。方法：通过高通量阵列,利用miRNAs芯片筛选差异性miRNAs,予生物信息学方法预测3’-非编码区(3’-UTR)区域含有CLOCK结合位点的差异性miRNA。荧光素酶报告基因系统等技术验证松果体中miR-182表达在CLOCK转录调控中的分子机制。结果：我们证明了miR-182调控了CLOCK基因(生物钟调控过程中的核心钟基因之一)的3’-UTR；并且在通过氧葡糖剥夺环境的原代培养松果体细胞上进一步验证了miR-182对CLOCK表达的调控作用。结论：miR-182在松果体组织中高表达,并且与CLOCK的表达相关。CLOCK是miR-182的目标靶基因,为缺氧缺血导致的松果体功能损害相关的节律紊乱的治疗提供了新的靶点
[Abstract]:Neonatal hypoxic ischemic encephalopathy (HIEs) is a common severe complication of neonatal asphyxia. However, the circadian rhythms in children with HIE of different degrees have not been paid attention to, and their mechanisms are less understood. Therefore, further study on sleep disorders in children with HIE of different degrees, and possible mechanisms of participation, can improve the treatment plan, improve the quality of life of the children, and alleviate the burden on the family. MiRNAs have important clinical value and significance. MiRNAs, as important gene expression regulators, are closely related to the occurrence and development of many diseases. However, the role of miRNAs in HIE and its mechanism have not been reported. In this study, a follow-up study of sleep status in patients with different degrees of HIE and a systematic study of pineal tissue in animal models were conducted. To explore the mechanism of the selected miR-182 in circadian rhythm abnormalities caused by pineal dysfunction after hypoxic-ischemic injury in order to provide a new target for the biotherapy of sleep disorders in children with HIE. Part I effects of mild and moderate HIE on sleep in children objective: this part aims to elucidate the characteristics of sleep disorder in children with non-severe perinatal hypoxic-ischemic injury. Methods: from March 2010 to August 2013, 128 newborns with a history of hypoxic-ischemic encephalopathy were admitted to the affiliated Children's Hospital of Suzhou University. The clinical data of 88 normal full-term newborns matched with age and sex in the same period were collected and the sleep status was evaluated by questionnaire. To explore the problem of sleep disorder in children with perinatal HIE of different degree by Wilcoxon symbol grade test. Results: through the investigation of sleep therapy in children with HIE of different degrees, it was found that the mild to moderate HIE children showed obvious sleep initiation and maintenance, sleep related breathing problems and circadian rhythm problems, compared with those of mild HIE children, compared with mild HIE children, the patients with mild and moderate HIE showed obvious sleep initiation and maintenance, sleep related breathing problems and circadian rhythm problems. The risk of sleep apnea in moderate HIE children was significantly higher than that in moderate HIE children. The risk of circadian rhythm disorder in mild HIE children was higher than that in moderate HIE children. Conclusion: the results of this study suggest that mild and moderate HIE children have different degrees of surface. Showing obvious sleep initiation and maintenance, The sleep related breathing problems and circadian rhythm problems provide a clinical basis for improving the long-term prognosis of children with HIE through the targeted treatment of sleep disorders. Part two the role of miR-182 in regulating the expression of CLOCK in pineal gland of neonatal rats after HIBD objective: to establish a model of hypoxic-ischemic brain injury in neonatal rats in vivo and in vitro. To analyze the mechanism of miR-182 in circadian rhythm disorder caused by pineal dysfunction. Methods: miRNAs microarray was used to screen the differential miRNAs.The bioinformatics method was used to predict the difference of miRNAs with CLOCK binding sites in the 3G-non-coding region. Luciferase reporter gene system and other techniques were used to verify the molecular mechanism of miR-182 expression in the CLOCK transcriptional regulation in the pineal gland. Results: we demonstrated that miR-182 regulates the expression of CLOCK gene (one of the core clock genes in the biological clock), and further demonstrated the role of miR-182 in the regulation of CLOCK expression in primary cultured pineal cells exposed to oxygen glucose-deprived environment. Conclusion the high expression of miR-182 in pineal tissue and its correlation with the expression of CLOCK. CLOCK is the target gene of miR-182, which provides a new target for the treatment of circadian disorder related to the damage of pineal function induced by hypoxia and ischemia.
【学位授予单位】：苏州大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R742

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