宁神安脏饮对失眠大鼠下丘脑生物钟基因表达的影响OA

Objective:To investigate effects of Ningshen Anzang Yin on expression of hypothalamic circadian clock genes in insomnia model rats.Methods:Eighty Sprague-Dawley(SD)rats were randomly divided into four groups blank control group,model group,Ningshen Anzang Yin group,and eszopiclone group,with 20 rats in each group.Except for the blank control group,the other groups were subjected to chronic unpredictable mild stress(CUMS)combined with intraperitoneal injection of para-chlorophenylalanine(PCPA)to establish an insomnia model.The Ningshen Anzang Yin group received 2.36 g/(kg·d)by gavage,the eszopiclone group received 0.32 mg/(kg·d)by gavage,while the blank and model group received an equal volume of 0.9％saline solution daily for 14 consecutive days.General condition and body weight were observed.Sleep latency and duration were assessed using pentobarbital sodium-induced sleep tests.Neuronal morphology in the hypothalamus was examined via hematoxylin-eosin(HE)staining.mRNA expression levels of CLOCK,BMAL1,Per1,and Cry1 in the hypothalamus were detected by qRT-PCR.Protein expression was analyzed by Western blot and immunohistochemistry.Results:Compared with the blank control group,model rats showed poor general condition,reduced body weight(P＜0.05),shortened sleep duration,and vacuolar degeneration of hypothalamic neurons.Compared with the model group,both the Ningshen Anzang Yin and eszopiclone group exhibited improved general condition,increased body weight(P＜0.05),prolonged sleep dura-tion,and ameliorated neuronal morphology.In the model group,mRNA and protein expression of CLOCK and BMAL1 was elevated,whereas that of Per1 and Cry1 was decreased(P＜0.01).Both treatment groups reversed these abnormalities(P＜0.05).The Ningshen Anzang Yin group significantly reversed the protein expression of CLOCK,Per1,and Cry1(P＜0.01),while the eszopiclone group improved CLOCK and Per1 protein expression(P＜0.01).Immunohistochemical analysis revealed enhanced positive expression of CLOCK,BMAL1,Per1,and Cry1 in both treated groups(P＜0.01).Conclusion:Ningshen Anzang Yin can improve behavioral status and sleep quality in in-somnia model rats,possibly through regulation of hypothalamic circadian clock gene expression.