中国神经再生研究(英文版)

中国神经再生研究(英文版) ›› 2021, Vol. 16 ›› Issue (10): 2071-2077.doi: 10.4103/1673-5374.308101

• 原著:脑损伤修复保护与再生 • 上一篇    下一篇

新生儿缺氧缺血性脑损伤后松果体miR-325在昼夜节律调节中的作用

  

  • 出版日期:2021-10-15 发布日期:2021-03-19
  • 基金资助:

    中国国家自然科学基金项目（81871193，81671532，81771625，81701490，81801505）；江苏省青年医学人才项目（QNRC2016763，QNRC2016758，QNRC2016762）；苏州市科技计划项目（SS201709）；江苏省自然科学基金面上项目（BK20180205）；姑苏卫生人才培训计划基金（GSWS2019049）；江苏省重点医学学科（ZDXKA2016013）； 江苏省妇幼健康研究项目（F201750）；苏州市儿科临床中心（Szzx201504）；苏州工业技术创新项目（SYS201765）；苏州科教卫生技项目（KJXW2018018）

The role of pineal microRNA-325 in regulating circadian rhythms after neonatal hypoxic-ischemic brain damage

Ning Sha1, 2, #, Hua-Wei Wang1, #, Bin Sun1, Min Gong1, Po Miao1, Xiao-Lu Jiang3, Xiao-Feng Yang1, Mei Li3, Li-Xiao Xu3, Chen-Xi Feng3, Yuan-Yuan Yang4, Jie Zhang5, Wen-Jing Zhu5, Yuan-Yuan Gao6, Xing Feng1, *, Xin Ding1, *#br#   

  1. 1Soochow Key Laboratory of Prevention and Treatment of Child Brain injury, Children’s Hospital of Soochow University, Suzhou, Jiangsu Province, China; 2Department of Pediatrics, Affiliated Huai’an Hospital of Xuzhou Medical University, Huai’an, Jiangsu Province, China; 3Pediatrics Research Institute, Children’s Hospital of Soochow University, Suzhou, Jiangsu Province, China; 4Department of Pediatrics, The First Affiliated Hospital of Soochow University (Dushuhu Branch), Suzhou, Jiangsu Province, China; 5Cambridge-SU Genomic Resource Center, Soochow University, Suzhou, Jiangsu Province, China; 6Clinical Laboratory, Children’s Hospital of Soochow University, Suzhou, Jiangsu Province, China.
  • Online:2021-10-15 Published:2021-03-19
  • Contact: Xin Ding, MD, PhD, dingxin@suda.edu.cn; Xing Feng, MD, xing_feng66@suda.edu.cn.
  • Supported by:
    This study was supported by the National Natural Science Foundation of China, Nos. 81871193 (to XD), 81671532 (to BS), 81771625 & 81701490 (to XF), 81801505 (to MG); Jiangsu Provincial Medical Youth Talent of China, Nos. QNRC2016763 (to XD), QNRC2016758 (to LXX), QNRC2016762 (to ML); the Science and Technology Project of Suzhou City of China, No. SS201709 (to XD); the Natural Science Foundation of Jiangsu Province of China, No. BK20180205 (to XD); the Training Program Foundation for Health Talents of Gusu of China, No. GSWS2019049 (to XD); the Jiangsu Provincial Key Medical Discipline of China, No. ZDXKA2016013 (to XF); the Jiangsu Province Women and Children Health Research Project of China, No. F201750 (to LXX); the Pediatric Clinical Center of Suzhou City of China, No. Szzx201504 (to XF); Suzhou Industrial Technology Innovation Project of China, No. SYS201765 (to LZ), and the Project of Suzhou Science, Education and Health and Technology, China, No. KJXW2018018 (to ML).

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摘要：

昼夜节律紊乱是新生儿缺氧缺血性脑损伤常见且易被忽视的症状。但是其潜在的分子机制仍然未知。作者既往研究通过缺氧缺血性脑损伤新生大鼠模型，揭示了松果体中miR-325上调可明显抑制褪黑素合成和昼夜节律调节的关键酶Aanat的产生。为了更好的揭示新生儿缺氧缺血性脑损伤与松果体miR-325及昼夜节律障碍发生的机制，①此次试验于2019年在苏州大学第一附属医院收集了缺氧缺血性脑损伤新生患儿及健康新生儿进行比较，发现循环miR-325与新生儿缺氧缺血性脑损伤患儿睡眠和昼夜节律问题的严重程度呈正相关；②以荧光素酶检测出LHX3是miR-325的直接下游靶标；③利用miR-325基因敲除小鼠证明，lhx3作为一种转录因子，通过依赖松果体miR-325的表达昼夜节律。进一步通过左颈总动脉双层结扎并低氧饲养2h建立新生儿缺氧缺血性脑损伤小鼠模型，显示其松果体LHX3mRNA含量下降，但敲除基因miR-325的新生小鼠则出现逆转迹象，新生儿缺氧缺血性脑损伤动物昼夜节律的行为改善依赖于miR-325和LHX3。④该实验阐明了miR-325-LHX3轴调节新生儿缺氧缺血性脑损伤昼夜节律紊乱的机制，并为治疗靶点提供了新的见解。临床试验于2015年7月20日经苏州大学儿童医院机构审查委员会批准（批准号2015028），动物实验于2016年1月15日经苏州大学医学院动物伦理委员会批准（批准号XD-2016-1）。

https://orcid.org/0000-0002-0387-3240 (Xin Ding)

关键词:

新生儿, 缺氧缺血性脑损伤, 昼夜节律, 松果体, 脑缺血, miRNA, 睡眠, 转录因子

Abstract: Circadian rhythm disorder is a common, but often neglected, consequence of neonatal hypoxic-ischemic brain damage (HIBD). However, the underlying molecular mechanisms remain largely unknown. We previously showed that, in a rat model of HIBD, up-regulation of microRNA-325 (miR-325) in the pineal gland is responsible for the suppression of Aanat, a key enzyme involved in melatonin synthesis and circadian rhythm regulation. To better understand the mechanism by which miR-325 affects circadian rhythms in neonates with HIBD, we compared clinical samples from neonates with HIBD and samples from healthy neonates recruited from the First Affiliated Hospital of Soochow University (Dushuhu Branch) in 2019. We found that circulating miR-325 levels correlated positively with the severity of sleep and circadian rhythm disorders in neonates with HIBD. Furthermore, a luciferase reporter gene assay revealed that LIM homeobox 3 (LHX3) is a novel downstream target of miR-325. In addition, in miR-325 knock-down mice, the transcription factor LHX3 exhibited an miR-325-dependent circadian pattern of expression in the pineal gland. We established a neonatal mouse model of HIBD by performing double-layer ligation of the left common carotid artery and exposing the pups to a low-oxygen environment for 2 hours. Lhx3 mRNA expression was significantly down-regulated in these mice and partially rescued in miR-325 knockout mice subjected to the same conditions. Finally, we showed that improvement in circadian rhythm-related behaviors in animals with HIBD was dependent on both miR-325 and LHX3. Taken together, our findings suggest that the miR-325-LHX3 axis is responsible for regulating circadian rhythms and provide novel insights into the identification of potential therapeutic targets for circadian rhythm disorders in patients with neonatal HIBD. The clinical trial was approved by Institutional Review Board of Children’s Hospital of Soochow University (approval No. 2015028) on July 20, 2015. Animal experiments were approved by Animal Care and Use Committee, School of Medicine, Soochow University, China (approval No. XD-2016-1) on January 15, 2016.

Key words: brain injury, circadian rhythm, hypoxic-ischemic brain damage, miRNA, neonate, pineal gland, sleep, transcription factor 

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