Changes Of Gut Microbiota In Sleep-restricted Mice And Its Relationship With Cognition And Emotion

ObjectivesTo explore the effects of sleep restriction on the ability of learning and memory,and gut microbiota in mice through animal experiments,and clarify the relationship between gut microbiota and emotion,and cognition,for from the mutual relationship between the host and gut microbiota to provide a scientific basis for explaining the psychological and mental problems caused by sleep disorders.MethodsThe subjects were healthy 8-week-old male C57BL/6J mice.The mice were randomly divided into normal control group(NC group),environmental control group(EC group)and sleep restriction group(SD group).The sleep restriction model was established by Modified Multiple Platform Method(MMPM).The experimental mice were subjected to sleep deprivation for 6h/d(8:00 ～14:00)in a sleep deprivation chamber,and the duration of sleep restriction was 8 weeks.Morris water maze test to assess learning and memory ability in mice.The open-field experiment could evaluate the anxiety state of mice.The concentrations of inflammatory cytokines were determined by Enzyme-Linked Immunosorbent Assay.The changes of gut microbiota in mice were analyzed by 16 S r RNA sequencing.Results(1)In the 8-week experiment,the weekly weight of the SD group were higher than that of the control groups,and the difference was statistically significant(all P<0.05).(2)The Morris water maze test showed that the escape latency of the SD group on the 2,3,4,and 5 days were significantly longer than that of the control groups(all P<0.05).The number of times the original platform was traversed and percentage of swimming time and the percentage of swimming distance in the target quadrant in the SD group significantly decreased(all P<0.05).These pictures of the path of the mice in the open field showed that the SD group mice were inclined to move around and in the four corners of the open field(indicating anxiety-like behavior),and the SD group with less distance in the central area of the open field,and the difference was statistically significant(all P<0.05).(3)The detection of pro-inflammatory cytokines showed that sleep restriction caused a 69.28%,61.18%,and 50.76 % increase in serum IL-1β,IL-6 and TNF-α in mice,and the difference was significant(all P<0.001).(4)16S r RNA sequencing analysis: the result of Beta diversity analysis showed that the R value of Anosim test was 0.392,P=0.016.Shannon index and Simpson index were different between the two groups and significantly decreased in the SD group(P=0.021 and P=0.010).LEf Se analysis showed that in the SD group,Tannerellaceae,Rhodospirillales,Alistipes,and Parabacteroides were significantly increased;while in the EC group,Ruminococcaceae(UCG＿013 and UCG＿010),Lachnospiraceae(NK4A136＿group and bacterium＿DW17)and Eubacterium＿xylanophilum＿group were more enriched.(5)Correlation analysis: the level of IL-1β in mouse serum was positively correlated with the abundance of Muribaculaceae(r=0.497,P<0.05),and negatively correlated with the abundance of Lachnospiraceae(r=-0.583,P<0.05).The level of TNF-α was positively correlated with the abundance of Erysipelotrichaceae,Burkholderiaceae,and Tannerellaceae(r=0.492,r=0.646,r=0.726,all P<0.05).The abundance of Akkermansiaceae and Rikenellaceae in mice were positively correlated with the total distance of the open field experiment and the distance to stay in the central area(r=0.699,r=0.735,r=0.637,r=0.836,all P<0.05).The abundance of Burkholderiaceae was negatively correlated with the total distance in the open field experiment(r=-0.679,P<0.05).Conclusions(1)Sleep restriction could induce cognitive impairment and anxiety-like behavior in mice.(2)Sleep restriction could increase the level of pro-inflammatory cytokines in serum of mice,and the inflammatory factors in the serum may be caused by the disorder of the microbiota.(3)Sleep restriction could affect the gut microbiota,significantly promote the increase of harmful bacteria such as Rhodospirillales,Alistipes,and the decrease of probiotics such as Lachnospiraceae,Ruminococcaceae in the intestines,and these microorganisms play an important role in mood regulation and cognitive development.The findings of this study provide new evidence for changes in the composition of gut microbiota in patients with sleep disorders and possible directions for the diagnosis and treatment of neuropsychiatric diseases.