睡眠干扰调控5-羟色胺2A受体表达抑制动物骨骼增长的机制及miRNA筛选研究  

Research of Sleep Disturbance Regulates 5-HT2A Receptor Expression to Inhibit Animal Bone Growth Mechanism and miRNA Screening

作　　者：卢承印 吕艳艳 谢艳 LU Chengyin;LYU Yanyan;XIE Yan(Luoyang Orthopedic-Traumatological Hospital of Henan Province,Luoyang 471002,Henan China;Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南省洛阳正骨医院/河南省骨科医院,河南洛阳471002 [2]河南中医药大学

出　　处：《中国中医骨伤科杂志》2022年第3期7-11,共5页Chinese Journal of Traditional Medical Traumatology & Orthopedics

基　　金：河南省中医药科学研究专项课题(2018ZY1023)。

摘　　要：目的:研究5-羟色胺2A受体在睡眠干扰中抑制动物骨骼增长的作用并筛选靶向5-羟色胺2A受体调控其表达的miRNA。方法:通过对小鼠进行睡眠干扰,观察其对小鼠体长增长、5-羟色胺2A受体表达、生长激素分泌的影响;同时通过对大鼠进行睡眠干扰,测量其对大鼠慢波睡眠、异相睡眠等各类型睡眠时长的影响;采用芯片法筛选睡眠干扰后引起小鼠体长差异表达的miRNA,利用RnaDB网站筛选调节5-羟色胺2A受体表达的miRNA,对得到的两组miRNA进行比较,得到重合的miRNA并进行qRT-PCR验证,从而筛选出通过对睡眠干扰引起5-羟色胺2A受体高表达抑制动物骨骼增长的miRNA。结果:睡眠干扰20 d后,明显抑制小鼠体长增长,同时显著增加小鼠脑组织5-羟色胺2A受体表达,降低体内生长激素分泌,与空白对照组比较差异有统计学意义(P<0.05)。同时,睡眠干扰能明显减少大鼠慢波睡眠和总睡眠时长(P<0.05),但在异相睡眠时长上无明显影响(P>0.05)。在miRNA筛选研究方面,芯片法共获得16个符合差异倍数的miRNA,其中表达上调13个,表达下调3个,利用RnaDH网站筛选与芯片法筛选重合的miRNA为miR-200c-3p,对其进行qRT-PCR法验证结果显示其确能引起睡眠干扰组与空白对照组差异表达,因此miR-200c-3p可能为睡眠干扰引起5-羟色胺2A受体高表达进而抑制动物骨骼增长的核心靶点之一。结论:睡眠干扰能够抑制动物骨骼增长,其机制可能与睡眠干扰引起5-羟色胺2A受体高表达,导致慢波睡眠减少,从而降低生长激素分泌有关,通过miRNA筛选及qRT-PCR验证显示,miR-200c-3p可能是其核心靶点之一。Objective:To study the efficacy of 5-hydroxytryptamine(5-HT) 2 A receptors in inhibiting bone growth in animals during sleep disturbance and to screen miRNA that target 5-HT2 A receptors to regulate their expression.Methods:The efficacy on the growth of mouse body length, 5-HT2 A receptor expression and growth hormone secretion was observed after sleep disturbance in mice, meanwhile, through sleep disturbance in rats.Its efficacy on rat slow-wave sleep and the influence of various types of sleep duration, such as heterogeneous sleep was measured were detected, the chip method was used to screen the miRNA that caused differential expression of mouse body length after sleep interference, and the RnaDB website was used to screen the miRNA that was regulated to the expression of 5-HT2 A receptors.For comparison, the overlapping miRNA were obtained and verified by qRT-PCR, so as to screen out miRNA that inhibited the growth of animal bones by causing high expression of 5-HT2 A receptors by disturbing sleep.Results:The body length growth of mice was significantly inhibited after sleep disturbance for 20 d.At the same time, the expression of 5-HT2 A receptor in the brain tissue of mice was significantly increased, and the secretion of growth hormone in the body was reduced.Compared with the blank control group, the difference was statistically significant(P< 0.05).At the same time, the slow-wave sleep and total sleep duration of rats significantly reduced by sleep disturbance(P<0.05), but it has no significant efficacy on the duration of paradoxical sleep(P>0.05).In terms of miRNA screening research, a total of 16 miRNA with different multiples were obtained by the chip method, of which 13 were up-regulated in expression and 3 were down-regulated.The verification results of qRT-PCR method showed that differential expression can be indeed caused between the sleep disturbance group and the blank control group.Therefore, miR-200 c-3 p may be one of the core targets for sleep disturbance to cause high expression of 5-HT2 A

关 键 词：睡眠干扰 慢波睡眠 生长激素 骨骼增长 5-羟色胺 

分 类 号：R-33[医药卫生]