Comparison and evaluation of three methods for preparing insomnia mouse models

Li Feifan; Zhang Yibo; Wang Jing; Zhu Jinqiang; Zheng Wenke

doi:10.12307/2026.749

Abstract：BACKGROUND:Stable animal insomnia models serve as crucial tools for investigating insomnia mechanisms and developing anti-insomnia drugs.The combination of chemical and physical methods provides an effective approach for establishing animal insomnia models. OBJECTIVE:To compare chemical methods(p-chlorophenylalanine),physical methods(water environment sleep deprivation method),and combined methods(p-chlorophenylalanine combined with water environment sleep deprivation method)for constructing insomnia mouse models,thereby exploring the optimal modeling method. METHODS:C57 mice were randomly divided by sex into the following groups:control(untreated),p-chlorophenylalanine,water platform,and p-chlorophenylalanine+water platform,with 12 mice per group(equal numbers of males and females).Mice in the p-chlorophenylalanine group received intraperitoneal injections of 400 mg/kg p-chlorophenylalanine suspension daily for 3 consecutive days;those in the water platform group were placed in a water environment sleep deprivation platform for 3 consecutive days;and those in the p-chlorophenylalanine+water platform group were placed in the sleep deprivation platform and received intraperitoneal injections of 400 mg/kg p-chlorophenylalanine suspension for 3 consecutive days.After 3 days of modeling,mice were subjected to the open field test,and the number of neurons in the hypothalamus and hippocampus was examined via Nissl staining.Serum levels of 5-hydroxytryptamine,gamma-aminobutyric acid,and dopamine were measured using enzyme-linked immunosorbent assay. RESULTS AND CONCLUSION:(1)Compared with the control group,male mice in the three model groups exhibited an upward trend in total distance,number of entries into the central area,average speed,and number of stand-up episodes.The most pronounced changes were observed in the p-chlorophenylalanine+water platform group(P<0.05 or P<0.01).Female mice in the three model groups also showed an upward trend in total distance,number of entries into the central area,average speed,and number of stand-up episodes.(2)All three modeling methods could reduce the number of neurons in the mouse hypothalamus and hippocampus,with the highest reduction and most severe neuronal damage observed in female mice in the p-chlorophenylalanine+water platform group.(3)Compared with female mice in the control group,changes in three serum indicators were more pronounced in the female mice in the p-chlorophenylalanine+water platform group.5-Hydroxytryptamine and gamma-aminobutyric acid levels were significantly reduced(P<0.05),while dopamine levels were significantly elevated(P<0.01).To conclude,the insomnia model established in female mice using p-chlorophenylalanine combined with the water platform is relatively more reliable and stable,providing an ideal animal model for insomnia research.