摘要
目的观察六味安神胶囊对老年睡眠剥夺斑马鱼模型的睡眠改善作用。方法60尾18月龄雌性斑马鱼随机分为空白组、模型组、六味安神组、褪黑素组,空白组斑马鱼正常灯光条件下饲养,其余组斑马鱼均采用持续光照3天构建睡眠剥夺模型,六味安神组斑马鱼在模型组基础上叠加六味安神胶囊水溶液0.00050 mg·mL^(-1)处理3天,褪黑素组斑马鱼在模型组基础上叠加褪黑素水溶液0.2 mg·mL^(-1)处理3天。3天后运用斑马鱼行为学系统检测各组斑马鱼静息时间,qRT-PCR检测各组斑马鱼周期蛋白1a(per1a)、周期蛋白2(per2)、昼夜运动输出周期1a(clock1a)、隐色素1b(cry1b)、5-羟吲哚乙酸(5-htiaa)和卵泡刺激素β(fshβ)基因的表达水平,Western blot方法检测各组斑马鱼雌激素受体α(Esrα)、Fshβ、促黄体生成素(Lh)表达水平,HE染色观察各组斑马鱼卵巢组织形态。结果模型组斑马鱼在观察的24 h中,与空白组斑马鱼相比,静息时间明显减少(P<0.01),给予六味安神胶囊和褪黑素干预后,斑马鱼的静息时间均明显增多(P<0.01)。与空白组斑马鱼对比,睡眠剥夺后的斑马鱼生物钟基因per1a、per2、clock1a、cry1b、5-htiaa、fshβmRNA表达量均呈现出下降趋势(P<0.05或P<0.01),而在给予六味安神胶囊干预后,per1、clock1a、cry1b、5-htiaa、fshβmRNA表达量均上调(P<0.05或P<0.01)。与空白组斑马鱼相比,模型组斑马鱼的Esrα、Lh蛋白表达量均上调(P<0.05),Fshβ蛋白表达量下调(P<0.05),给予六味安神胶囊干预后,上述指标未见明显变化。空白组斑马鱼的卵巢组织细胞形态正常,初级卵母细胞的数量较多,而模型组斑马鱼的卵巢组织细胞形态出现破坏,初级卵母细胞的数量减少,给予六味安神胶囊和褪黑素干预后,斑马鱼卵巢组织的细胞形态虽仍有不同程度的破坏,但整体较完整,初级卵母细胞增多。结论老年斑马鱼的失眠可能是多因素导致的,六味安神胶
Objective Observe the effect of Liuwei Anshen Capsule on sleep improvement of aged sleep deprivation zebrafish model.Methods Sixty 18-month-old female zebrafish were randomly divided into blank group,model group,Liuwei Anshen group,and melatonin group.The zebrafish in the blank group were raised under normal lighting conditions,and the other three groups were constructed with continuous lighting for 3 days.Zebrafish in Liuwei Anshen group were treated with Liuwei Anshen capsule water solution 0.00050 mg·mL^(-1) for 3 days on the basis of model group,and zebrafish in melatonin group were treated with melatonin aqueous solution 0.2 mg·mL^(-1) on the basis of model group for 3 days.After 3 days,the zebrafish behavior system was used to detect the resting time of zebrafish in each group.qRT-PCR method was used to detect cyclin 1a(per1a),cyclin2(per2),circadian motor output cycle 1a(clock1a),cryptochrome 1b(cry1b),and 5-hydroxyindoleacetic acid(5-htiaa)in each group of zebrafish and follicle-stimulating hormone beta(fshβ)gene expression levels.Western blot was used to detect the expression levels of estrogen receptorα(Esrα),Fshβand luteinizing hormone(Lh)in zebrafish in each group.HE staining was used to observe the ovary of zebrafish in each group.Results Compared with the zebrafish in the blank group,the resting time of the zebrafish in the model group decreased significantly(P<0.01)during the 24 hours of observation.After the intervention of Liuwei Anshen Capsule and melatonin,the resting time of the zebrafish was significantly increased.(P<0.01).Compared with the zebrafish in the blank group,the mRNA expressions of zebrafish circadian clock genes per1a,per2,clock1a,cry1b,5-htiaa,and fshβall showed a downward trend after sleep deprivation(P<0.05,P<0.01).After the intervention of Liuwei Anshen Capsules,the mRNA expressions of per1,clock1a,cry1b,5-htiaa and fshβwere all up-regulated(P<0.05,P<0.01).Compared with the zebrafish in the blank group,the protein expressions of Esrαand Lh in the zebrafish of the mod
作者
冉思邈
鲁海菲
叶子靖
王瑛
段彩虹
王平
Ran Simiao;Lu Haifei;Ye Zijing;Wang Ying;Duan Caihong;Wang Ping(Institute of Geriatric Medicine,Hubei University of Chinese Medicine,Wuhan 430000,China;Chongqing Huasen Pharmaceutical Co.LTD,Chongqing 401121,China)
出处
《世界科学技术-中医药现代化》
CSCD
北大核心
2023年第11期3728-3735,共8页
Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology
基金
国家中医药管理局中医药传承与创新“百千万”人才工程(歧黄工程)——国家中医药领军人才支持计划(国中医药人教发[2018]12号),负责人:王平
关键词
失眠
睡眠剥夺
老年斑马鱼
六味安神胶囊
Insomnia
Sleep deprivation
Aged-zebrafish
Liuwei Anshen Capsule
