慢性不完全睡眠剥夺对小鼠卵巢储备功能的影响  

Impact of chronic incomplete sleep deprivation on ovarian reserve function in mice

作　　者：潘烨栋 周晟 王严慧 张杨 常柳燕 杨素荣 Pan Yedong;Zhou Sheng;Wang Yanhui;Zhang Yang;Chang Liuyan;Yang Surong(Department of Pharmacology,School of Basic Medical Sciences,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学基础医学院药理学系,上海200032

出　　处：《中华生殖与避孕杂志》2024年第6期574-578,共5页Chinese Journal of Reproduction and Contraception

基　　金：国家自然科学基金面上项目(32371033,32170983);上海市自然科学基金面上项目(21ZR1408000);复旦大学上海医学院卿枫学者本科生创新创业项目(QF2134)。

摘　　要：目的探讨慢性不完全睡眠剥夺(sleep deprivation,SD)对小鼠卵巢储备功能的影响及其可能作用机制。方法16只SPF级7~8周龄雌性C57BL/6小鼠,适应性饲养1周后,简单随机化分为对照组和SD组,每组8只,其中SD组小鼠每天7∶00~10∶00采用剥夺杆进行慢性不完全SD,共计40 d。在实验开始前与结束前的9 d内每天进行阴道脱落细胞涂片检查,记录动情周期。于第40天通过酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)试剂盒检测血清雌二醇、孕酮、卵泡刺激素(follicle-stimulating hormone,FSH)、促黄体生成素(luteinizing hormone,LH)和褪黑素(melatonin,MT)水平。取卵巢,计算卵巢指数,并通过切片苏木精-伊红染色计数原始卵泡数量。免疫组织化学方法检测灌注取脑后小鼠视交叉上核(suprachiasmatic nucleus,SCN)脑区神经元c-Fos表达情况。结果相比于对照组,SD组小鼠动情周期出现紊乱趋势。SD组小鼠雌二醇[(20.19±3.67)ng/L]、孕酮[(2.88±0.53)μg/L]、FSH[(13.42±2.36)U/L]水平明显低于对照组小鼠[(24.66±2.15)ng/L,P=0.010;(3.43±0.49)μg/L,P=0.049;(17.01±1.49)U/L,P=0.003]。SD组小鼠LH和MT水平低于对照组小鼠,但差异均无统计学意义(均P>0.05)。SD组小鼠卵巢原始卵泡数量与对照组小鼠相比,差异无统计学意义(P>0.05),但是SD组卵母细胞形态不佳,颗粒细胞排列紊乱,闭锁卵泡数量减少,卵巢纤维化明显。免疫组织化学染色观察到SD组小鼠SCN区神经元c-Fos蛋白表达上调。结论连续40 d每天进行3 h SD后小鼠卵巢储备功能受损,可能与SCN过度激活有关。Objective To explore the effect of chronic incomplete sleep deprivation(SD)on ovarian reserve function in mice and its potential mechanisms.Methods Sixteen 7-8 week old female C57BL/6 SPF mice were randomly divided into control and SD groups(n=8 per group)after one week of acclimatization.The mice in the SD group were treated with SD from 7:00 to 10:00 using a deprivation rod for a total of 40 d.Before the first day and the last day of the experiment,vaginal smears were collected daily for 9 d to record and evaluate the estrous cycle.On the last day of the experiment,serum levels of estradiol,progesterone,follicle-stimulating hormone(FSH),luteinizing hormone(LH),and melatonin(MT)were measured by enzyme-linked immunosorbent assay(ELISA).The ovaries were dissected to calculate the ovarian index and count the number of primordial follicles by hematoxylin-eosin(HE)staining.Cardiac perfusion was performed to get the brains and the expression of c-Fos protein was observed in the suprachiasmatic nucleus(SCN)by immunohistochemistry.Results Compared with control group,SD group had a tendency of estrous cycle disorders.Hormone levels of estradiol[(20.19±3.67)ng/L],progesterone[(2.88±0.53)μg/L],and FSH[(13.42±2.36)U/L]in the SD group were significantly lower than those in control group[(24.66±2.15)ng/L,P=0.010;(3.43±0.49)μg/L,P=0.049;(17.01±1.49)U/L,P=0.003].Hormone levels of LH and MT in the SD group were lower than those in control group,but without statistical significances(all P>0.05).There was no significant change in the number of primordial follicles between the two groups(P>0.05).However,the oocyte morphology was poor,the granulosa cells were disorderedly arranged,the number of atretic follicles was decreased,and ovarian fibrosis was obvious in the SD group.Immunohistochemical staining showed an upregulation of c-Fos protein expression in the SCN of the SD group.Conclusion Continuous 3 h SD for 40 d impairs ovarian reserve function in mice,possibly related to excessive activation of the SCN.

关 键 词：睡眠剥夺 卵巢储备功能 动情周期 

分 类 号：R-332[医药卫生]