Evaluation study on zebrafish insomnia model by illumination method and combined illumination and caffeine method
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Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, affiliated to Shanghai University of Traditional Chinese Medicine

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    Abstract:

    【】 Objective: to establish different sleep deprivation models using zebrafish to provide more reproducible and practical modeling reference solutions for basic research on insomnia. Methods: Zebrafish insomnia model was induced by two interventions: continuous light (150 lux) and light plus caffeine, and the zebrafish were randomly divided into control, light, combined and caffeine (100 μmol/l) groups. The difference in locomotor ability of zebrafish in each group was observed by using open field experiment and circadian rhythm behavioral experiment; the expression and secretion of related sleep genes and 5-HT neurotransmitter were detected by using Qpcr technology and Elisa technology. Results: 1. The sleep time of zebrafish in the light group was significantly reduced compared with that of the blank control group (P=0.01), and there was no statistically significant difference between the sleep time of zebrafish in the combined group and the caffeine group (see Table 1); the resting time of zebrafish during the daytime in the light group was significantly reduced compared with that of the blank control group (P<0.01), and the resting time during the daytime of zebrafish in the combined group and the caffeine group was increased (P<0.01) (Table 3). There was no statistically significant difference in the black light resting time of zebrafish in all groups (Table 3); the daytime moving distance of zebrafish in the light group was significantly increased compared with that of blank control group (P < 0.01), and the 10-h activity distance of zebrafish in the light group in the dark day was significantly increased compared with that of blank control group (P < 0.01), and there was no statistically significant difference in the dark day activity distance of zebrafish in the combined and caffeine groups (Table 4); and the sleep rounds of zebrafish in the light group was blank control group was significantly reduced (P < 0.01), and the sleep rounds of zebrafish in the combined and caffeine groups were significantly increased (P < 0.01) compared to the blank control group (Table 2).2. The percentage of swimming distance in the central region of the light group was reduced (P < 0.05) compared to the blank group, while the percentage of swimming distance in the central region of the caffeine group was significantly reduced (P < 0.01) compared to the light group (see Figure 6); the light group The percentage of swimming time in the central region was lower than that in the blank group (P < 0.05), while the percentage of swimming time in the central region was significantly lower in the caffeine group than that in the light group (P < 0.01) (Fig. 7).3. The expression of HTR1aa mRNA level in zebrafish in the light group was higher than that in the blank group at 6:00 a.m. and 12:00 p.m. (P < 0.05) (Figs. 10 and 11), and the expression of There was no statistically significant difference in the expression of HTR1ab mRNA levels between the light group and the combined group (P > 0.05) (Figs. 12 and 13); compared with the blank group, the amount of 5-HT secretion in the light group was reduced in both groups at 6:00 a.m. (P < 0.01), and at 12:00 p.m. the amount of 5-HT was reduced in both the light and combined groups (P < 0.01), and the amount in both groups remained lower than that in the blank group at 6:00 p.m. (P<0.01).

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  • Received:June 17,2024
  • Revised:August 19,2024
  • Adopted:November 29,2024
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