昼夜节律紊乱对雄性青少年小鼠下丘脑和睾丸的影响  

Effects of circadian rhythm disturbances on the hypothalamus and testis in male adolescent mice

作　　者：刘宇 刘召平 蒲建林 向波 刘可智 蔡端芳 邬建飞 Liu Yu;Liu Zhaoping;Pu Jianlin;Xiang Bo;Liu Kezhi;Cai Duanfang;Wu Jianfei(Zigong Institute of Brain Science,The Zigong Affiliated Hospital of Southwest Medical University,Zigong 643020;Dept of Laboratory,The Zigong Affiliated Hospital of Southwest Medical University,Zigong 643020;Dept of Psychiatry,Affiliated Hospital of Southwest Medical Universit,Luzhou 646000)

机构地区：[1]西南医科大学附属自贡医院自贡市脑科学研究院,自贡643020 [2]西南医科大学附属自贡医院检验科,自贡643020 [3]西南医科大学附属医院精神科,泸州646000

出　　处：《安徽医科大学学报》2024年第10期1802-1808,共7页Acta Universitatis Medicinalis Anhui

基　　金：四川省科技计划项目(编号:2023NSFSC0124);四川省自贡市科技计划项目(编号:2023YKY11、2023-NKY-02-13、2023-NKY-02-14);四川省自贡市卫生健康委员会科研课题(编号:22yb001)。

摘　　要：目的探讨昼夜节律紊乱(CRD)对青少年小鼠下丘脑神经元变化和睾丸功能的影响。方法选取30只SPF级1月龄C57雄性小鼠,随机分为Control组和CRD组,每组15只,Control组保持12 h暗/12 h明的循环光照,CRD组保持24 h光照,造模共计61 d。统计各组小鼠的生长曲线;利用高架十字迷宫和旷场实验检测小鼠行为变化;利用尼氏染色检测神经元形态;利用免疫荧光染色和Western blot技术检测离子钙结合适配器分子1(Iba1)和神经元特异性核蛋白(NeuN)在下丘脑的分布和表达;利用RT-qPCR检测睾酮合成相关基因类固醇激素合成急性调节蛋白(StAR)和3β-羟基类固醇脱氢酶-1(HSD3B1)以及生精相关基因配子生成素结合蛋白2(GGNBP2)和无精症缺失样(DAZL)在睾丸中的表达情况。结果在第61天时,CRD组小鼠体质量显著高于Control组。高架十字迷宫中,CRD组小鼠进入开臂时间、开臂次数和开臂时间百分比都显著少于Control组小鼠;旷场测试中,CRD组小鼠在中心区停留时间和进入中心区频率都显著少于Control组小鼠。尼氏染色结果显示CRD组小鼠下丘脑阳性细胞数量显著少于Control组小鼠;免疫荧光和Western blot结果显示CRD组小鼠下丘脑的Iba1蛋白表达上调,NeuN蛋白表达下调。RT-qPCR结果显示CRD组小鼠睾丸中HSD3B1的表达显著低于Control组小鼠,GNBP2和DAZL的表达显著低于Control组小鼠。结论CRD处理不仅会导致青少年小鼠出现焦虑抑郁样行为,同时还会降低雄性青少年小鼠生精能力。Objective To explore the mechanisms of circadian rhythm disorder(CRD)on behavior and testicular spermatogenic capacity in adolescent mice.Methods Thirty SPF grade C57 mice were selected and randomly divided into the control and CRD groups with 15 mice in each group.The control group kept 12 h dark/12 h bright circulating light,and the CRD group kept 24 h light.The trial lasted for 61 days.The growth curves of mice in each group were counted;the elevated plus maze test and open field test were performed to detect mice behavior;neuronal morphology was visualized by Nissl staining.The distribution of ionized calcium-binding adapter molecule 1(Iba1)and neuron-specific nuclear protein(NeuN)in the hypothalamus were detected by immunofluorescence and Western blot.Expression of testosterone synthesis-related genes steroidogenic acute regulatory protein(StAR)and hydroxy-delta-5-steroid dehydrogenase,3 beta-and steroid delta-isomerase 1(HSD3B1)and spermatogenesis-related genes gametogenetin binding protein 2(GGNBP2)and deleted in azoospermia-like(DAZL)were determined by RT-qPCR.Results The weight of the CRD group was significantly higher than that of control group at 61 days;in the elevated plus maze test,the time,frequency,and percentage of time in the open arm of the CRD group were significantly less than those of the control group;in the open field test,there was no significant difference in movement distance between the two groups;however,the residence time of the central area in the CRD group was significantly less than that in the control group;the frequency of entering the central area in the CRD group was significantly less than that in the control group.Nissl staining results showed that the positive cells in the CRD group were significantly lower than the control group.Immunofluorescence and Western blot results showed that Iba1 protein expression was up-regulated and NeuN protein expression was down-regulated in the hypothalamus of the CRD group.In the RT-qPCR experiment,the expression of HSD3B1 in the CRD group

关 键 词：C57小鼠 昼夜节律紊乱 抑郁症 下丘脑 睾丸 

分 类 号：R742[医药卫生—神经病学与精神病学]