| Background:With the development of sleep medicine,researchers pay more and more attention to the relationship between sleep and environment,which leads to the establishment and development of sleep medicine in cold areas.Studies have shown that people in cold regions have serious sleep quality problems,which may be related to the geographical conditions,climatic factors and special living habits of people in cold areas.Sleep medicine in cold areas in China is in its infancy,so it is urgent to carry out a large sample epidemiological investigation and study on the mechanism of sleep disorders in cold areas.Objective:1.Through the investigation of sleep quality,CCMQ and living habits of middle-aged people in cold areas,the factors affecting sleep quality of middle-aged people in cold areas were analyzed.2.Through the intervention of cold factors and PCPA drugs in different groups of rats,the effects of cold factors on sleep quality and gene expression were analyzed.3.To explore the mechanism of acupuncture on sleep quality of insomnia rats in cold condition by comparing the effect of acupuncture on the content of serum IL-1 β in insomnia rats with cold factors and PCPA insomnia rats.Method:1.The Pittsburgh Sleep quality Index(PSQI),Constitution in Chinese Medicine Questionnaires and living habits were designed as questionnaires to investigate the middle-aged people in cold areas.A total of 605 valid questionnaires were collected,the information was collected and sorted out,and the statistical software SPSS 25.0 was used for data analysis.The measured values of PSQI were used as dependent variables,gender,exercise habits,daily nap time,work type,daily use of electronic equipment,BMI classification,seasonal factors and CCMQ were assigned as independent variables.The correlation between the total score of PSQI and gender,type of work,exercise habit,nap habit,duration of use of electronic equipment,BMI classification and seasonal influencing factors was analyzed by Person correlation analysis.The correlation equation between each factor and PSQI total score was constructed by multiple linear regression analysis.2.72 adult rats were randomly divided into 4 groups: room temperature group(n = 12),insomnia model group(n = 24),cold group(n = 12)and cold insomnia group(n = 24).The feeding conditions of the room temperature group and insomnia model group were as follows: normal ventilation,constant temperature(21 ±1 ℃),environmental humidity 40%-60%,automatic light and shade control(LRHD is 12:12,light period 8:00-20:00,dark period20:00-8:00),animals drink freely,in order to prevent the rat body weight from growing too fast,the daily diet is 6g feed / 100 g rat body weight.The feeding conditions of cold group and cold insomnia group were the same as those of room temperature group and insomnia model group except that they were placed in a constant temperature refrigerator of-10℃ from 18:00 to 19:30every day.After feeding for 15 days,the general behavior of rats was observed and their body mass was measured every day.On the 16 th day,insomnia model group and cold insomnia group were intraperitoneally injected with PCPA(350mg/kg)to establish sleep deprivation model,and the other groups were intraperitoneally injected with the same amount of normal saline for 3 days.The general behavior and the changes of body mass of rats in each group before and after modeling were observed and recorded.The differences of anxiety and depression in each group were compared by behavioral experiment(Open field test).The content of 5-HT in serum was determined by enzyme-linked immunosorbent assay(ELISA).3.Three rats were randomly selected from room temperature group and cold group,and the brain tissues were taken for transcriptome analysis.The samples were sequenced,screened and quality controlled.Significant differentially expressed genes were found.GO enrichment analysis and KEGG enrichment analysis of differentially expressed genes were carried out.4.Half of the rats were randomly selected from the insomnia model group to form the insomnia acupuncture group,and half of the rats from the cold insomnia model group were selected to form the cold insomnia acupuncture group.The insomnia acupuncture group and the cold insomnia acupuncture group were treated with the same acupuncture method: bilateral "Shenmai points" and "Zhaohai acupoints" were selected.0.18*13mm acupuncture needle was injected into the bilateral "Shenmai" and "Zhaohai" points of the rats at a fixed time every day,and the needle was retained for 20 min each time for 7 days.Operation requirements: first replenish "Zhaohai acupoint" and then purge "Shenmai acupoint",twist 1min,retain needle 20 min.The rats in the room temperature group,the cold group,the cold insomnia model group and the insomnia model group were given equal amount of grasping stimulation at the same time every day.The changes of general behavior and body mass of rats in room temperature group,insomnia model group,insomnia acupuncture group,cold group,cold insomnia model group and cold insomnia acupuncture group were observed before and after treatment,and the content of serum IL-1 β was determined by enzyme-linked immunosorbent assay(ELISA).Results:1.Investigation on sleep status of middle-aged people in cold cities: 605 valid questionnaires were analyzed and sorted out,of which 207(46.1%)participants had sleep quality problems and 16(3.6%)had serious sleep quality problems.Person correlation analysis showed that exercise habits,seasonal factors,daily use of electronic equipment and CCMQ were correlated with the total score of PSQI(P<0.05),while gender,type of work,length of daily nap and BMI grade were not correlated with the total score of PSQI(P >0.05).Multiple linear regression analysis simulates the correlation equation of various factors: PSQI measured value = 3.152-0.732 * daily exercise habits +0.228 * seasonal factors + 0.389 * daily use of electronic equipment + 0.423 *CCMQ.2.The effect of cold factors on sleep quality of rats and the difference of sleep quality between rats and PCPA: there was no significant difference in body mass among groups.After 15 days of feeding,the body weight of rats in cold group and cold insomnia group decreased significantly(P < 0.05),and the hair color was gray,messy,irritable and sleep time decreased,while the hair color of rats in room temperature group and insomnia model group was white,smooth,emotional stable and body mass increased steadily.24 hours after the establishment of PCPA model,the content of serum 5-HT in cold group was significantly lower than that in room temperature group(P < 0.001).The content of serum 5-HT in insomnia model group was significantly lower than that in room temperature group(P < 0.001).The content of serum 5-HT in cold insomnia model group was significantly lower than that in normal temperature blank group,cold blank group and normal temperature model group.3.Transcriptome analysis of the differences in gene expression between cold and normal rat brain tissues: screening and comparison showed that the significant differences were Cartpt,Trh,Rxrg and B9d1,in which the expression of Cartpt,Trh and Rxrg genes was down-regulated and B9d1 gene expression was up-regulated.4.Effect of acupuncture on sleep quality of insomnia rats with cold factors: after 7 days of acupuncture treatment,the body mass of rats in insomnia acupuncture group and cold insomnia acupuncture group increased significantly,which was significantly different from that in room temperature group,cold group,insomnia model group and cold insomnia model group(P <0.05).The content of serum IL-1β in insomnia acupuncture group and cold insomnia acupuncture group was significantly lower than that in other groups,but there was no significant difference in serum IL-1β content between the two groups.Conclusion:1.46.1% of the middle-aged people in cold areas have sleep quality problems,and 3.6% have serious sleep quality problems.The sleep quality of middle-aged people in cold areas is affected by exercise habits,time of use of electronic equipment,seasonal factors,CCMQ and other factors.There was a significant positive correlation between exercise and sleep quality of middle-aged people in cold areas,and a negative correlation between the use of electronic equipment and sleep quality of middle-aged people in cold areas.In terms of seasonal factors,winter has a great influence on the sleep quality of middle-aged people in cold areas.In terms of CCMQ,the sleep quality of middle-aged people with qi deficiency in cold areas is the worst.2.Cold condition can significantly reduce the sleep quality of rats.The effect of cold condition on sleep quality is related to the down-regulation of Cartpt,Trh and Rxrg gene expression and the up-regulation of B9d1 gene expression.3.Acupuncture can effectively improve the sleep quality of PCPA insomnia model rats and PCPA insomnia rats under cold conditions,and the improvement mechanism may be related to IL-1β cytokines. |
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