Effects of Gengxin Decoction(更欣汤)on Oestrogen Receptors and Orexin A in the Nucleus Accumbens of the Hypothalamus Nodularis Papillaris in Perimenopausal Insomnia Model Mice
Objective To explore the possible mechanism of Gengxin Decoction(更欣汤)in the treatment of perimenopausal insomnia from the perspective of the expression of oestrogen receptor and orexin A(OXA)in hypothalamic tuberomammillary nucleus(TMN).Methods Female ICR mice with regular estrous cycle were selected to estab-lish perimenopausal insomnia mice models by injecting 4-vinylcyclohexene diepoxide(VCD)combined with para-chlorophenylalanine(PCPA)suspension intraperitoneally.After successful modeling,75 mice models were randomly divided into model group,zopiclone group and low-,middle-,and high-dose Gengxin Decoction,with 15 mice in each group,and 15 normal mice were set as blank group.On the next day after successful modelling,mice in the low-,middle-,and high-dose Gengxin Decoction were given Gengxin Decoction granules 1.15,2.3 and 4.6 g·kg-1·d-1 by gavage at 9:00 AM every day,mice in the zopiclone group were given zopiclone suspension 1.13 mg·kg-1·d-1 by gavage at the same time point,and mice in the blank and model groups were given distilled water 1 ml·kg-1·d-1 by gavage at the same time point,for 14 days in all groups.During the experimental period,general conditions of the mice in each group,such as daytime activities,mental state,diet and coat colour,were observed;twenty-four hours after the final intervention,the sleep latency and sleep duration of mice were detected by sodium pentobarbital synergy test,and the levels of serum estradiol(E2)、progesterone(P)、follicle stimulating hormone(FSH)and OXA protein in TMN of hypothalamus were detected by ELISA,and the protein optical density values of estrogen receptor α(ERα),OXA and ERβ in TMN was detected by immunofluorescence staining.Results The circadian rhythm of mice was regular in the blank group;the circadian rhythm of mice was disordered in the model group,and they remained active during the day and night;the mental state of mice in each group of Gengxin Decoction was improved compared with that before,which showed that the daytime activity was less than before,and the coat color was slightly yellow but shiny,and the mental state of mice was significantly improved in the middle-dose Gengxin Decoction;the zopiclone group was less active during the day than before,had rough,less shiny hair,and was more alert.Compared with the blank group,the sleep latency of mice in the model group prolonged,the sleep duration shortened,serum E2 and P decreased,serum FSH and the expression of OXA protein in TMN region increased,the optical density of ERα and ERβ in TMN region decreased,and the optical density of OXA increased(P＜0.05 or P＜0.01).Compared with the model group,the sleep latency of the mice in the low-,middle-,and high-dose Gengxin Decoction groups and the zopi-clone group shortened,the sleep duration prolonged(P＜0.01);the serum E2 and P in the middle-,and high-dose Gengxin Decoction groups increased,FSH and the expression of OXA protein in TMN area decreased(P＜0.05);the optical density values of ERα in TMN area increased in the medium-dose group,and the optical density value of OXA decreased in low-and middle-dose Gengxin Decoction groups,and the optical density values of ERβ in TMN area increased in low-,middle-,and high-dose Gengxin Decoction groups and the zopiclone group(P＜0.05).Compared with the zopiclone group,mice in the low-dose Gengxin Decoction group had longer sleep latency and shorter sleep duration,and mice in the middle-and high-dose Gengxin Decoction groups had higher serum E2 level and ERβ optical density value in the TMN area,and lower serum FSH and OXA level in the TMN area(P＜0.05).Compared with the low-dose Gengxin Decoction group,the sleep latency of mice in the middle-and high-dose Gengxin Decoction groups shortened,and the serum FSH level reduced(P＜0.01).Conclusion Gengxin Decoction may inhibit arousal and improve perimenopausal insomnia by modulating the distribution of estrogen receptors and OXA in the TMN region,and inhibiting the overactivation of the orexin system in the TMN region.
NETL
NSTL
万方数据
