| Objective:This study employed bibliometrics to comprehensively assess the current state of sleep deprivation research,and integrated data mining techniques to optimize the selection of acupoints for electroacupuncture intervention targeting "Adjust Zang and Arouse Spirit" in sleep-deprived individuals.By analyzing the effects of this intervention on sleep quality,daytime sleepiness,memory function,as well as anxiety and depression levels before and after treatment,we aimed to provide clinical evidence regarding the efficacy of "Adjust Zang and Arouse Spirit" electroacupuncture for managing sleep deprivation.Furthermore,by investigating changes in behavior,intrabrain clock genes expression patterns and Glu/GABA-Gln metabolic loop alterations through administering "Adjust Zang and Arouse Spirit" electroacupuncture on sleep-deprived rats,we sought to elucidate its underlying mechanism.Methods:1 Utilizing VOSviewer,Citespace,and similar software,we conducted a visual analysis of 16,814 publications on sleep deprivation in the WOS database,examining factors such as the literature count,country,author,and keywords.This horizontal analysis aimed to evaluate the publications,assess literature utilization rates,and gain a scientific understanding of the current research landscape surrounding sleep deprivation.Additionally,a vertical analysis was conducted to delve into the keywords within the published literature,revealing clusters that highlight the primary research directions and hotspots in the field of sleep deprivation.2 A computer-based search was conducted on databases such as Web of Science,Pub Med,China National Knowledge Internet(CNKI),Wanfang Database,and VIP Journal Database,retrieving clinical research literature published between the establishment of these databases and 2023 specifically on the acupuncture treatment of insomnia,both in English and Chinese.An Excel spreadsheet was used to record acupuncture prescriptions,and a pivot table was created for descriptive statistical analysis of the frequency,meridian attribution,and specific acupoints used.For further analysis,the R4.3.1 software was employed.Association rule analysis was conducted using the arules package and the apriori algorithm,with a support level of ≥0.05 and a confidence level of ≥0.9.The arules Viz package was utilized for visualization of the analysis results.Additionally,the psych package within Rstudio software was used to calculate Spearman’s rank correlation coefficients,while the corrplot package visualized these coefficients.Hierarchical clustering was conducted to comprehensively analyze the patterns and core combinations of acupoint selection in acupuncture treatments for insomnia.3 A total of 69 subjects were recruited for a study on sleep deprivation,with 36 subjects in the control group and 33 subjects in the treatment group.The subjects were intervened with "Adjust Zang and Arouse Spirit" electroacupuncture,which lasted for 30 minutes per day.Each course of treatment consisted of 6 days of intervention followed by 1 day of rest,and the treatment was continuously applied for 4 courses.The subjects were evaluated and statistically analyzed using the Pittsburgh Sleep Quality Index(PSQI),Epworth Sleepiness Scale(ESS),Self-Rating Anxiety Scale(SAS),Self-Rating Depression Scale(SDS),Hamilton Anxiety Scale(HAMA),and Hamilton Depression Scale(HAMD)to assess the clinical efficacy of "Adjust Zang and Arouse Spirit" electroacupuncture intervention for sleep deprivation.4 By observing the general condition and behavioral changes(Morris water maze,open field test)of rats in each group,we aimed to determine the effects of "Adjust Zang and Arouse Spirit" electroacupuncture on weight,learning and memory ability,as well as anxiety and depressive mood in rats with sleep deprivation.Techniques such as rt-PCR,WB,ELISA,IHC,and IF were utilized to detect the expression levels of Clock,Bmal1,Cry1,and Per2 m RNA and protein in the SCN of rats in each group.Additionally,the contents of Glu,GABA,GABAARα1,GAD,and GS in the hypothalamus were measured,and the Glu/GABA ratio was calculated.These data were analyzed to assess the impact of "Adjust Zang and Arouse Spirit" electroacupuncture on the Clock,Bmal1,Cry1,and Per2 circadian clock genes and the Glu/GABA-Gln metabolic pathway in the SCN of sleep-deprived rats.Results:1 A total of 20,055 documents were retrieved,and 16,814 documents were obtained after excluding non-original and non-review literature.These documents included 135countries/regions,6,843 research institutions,and 3,348 academic journals.Among them,the country with the most publications was the United States(7,617 articles),the institution was the University of California System(863 articles),the author was Tufik,Sergio(199articles),and the journal was "SLEEP"(1,750 articles).The top 10 high-frequency keywords were: "sleep deprivation"(6,351),"deprivation"(4,944),"performance"(2,200),"duration"(1,139),"brain"(1,126),"rem sleep"(1,100),"stress"(820),"fatigue"(786),"risk"(783),"impact"(733).The manifestations,hazards,impacts,and brain-related mechanisms of sleep deprivation were studied.2 A total of 732 acupoint prescriptions were recorded,including 219 acupoints and a total frequency of 5,271 times.Shenmen acupoint had the highest frequency of 441 times(8.37%),while the highest frequency meridian was the foot solar bladder meridian with 867times(16.44%).There were 52 specific acupoints involved,with a total frequency of 1,753 times,and the original acupoints(12 acupoints)had the highest frequency of 1,001 times(19.19%).The core prescriptions of acupoint combinations obtained from association rules were Xinshu-Pishu and Shenmai-Zhaohai.In the cluster analysis,the Five Zang Organs Shu acupoints(Pishu,Xinshu,Ganshu,Shenshu,Feishu)in Group 2 were the main core clustering group.3 Clinical Effect of "Adjust Zang and Arouse Spirit" electroacupuncture on Sleep Deprivation Subjects.A statistical analysis was conducted on the PSQI,ESS,and WMS scores of sleep deprivation subjects before and after treatment.After treatment,the PSQI and ESS scores of the sleep deprivation subjects decreased,while the WMS score increased.There were statistically significant differences in PSQI,ESS(p<0.01),and WMS scores(p<0.05),indicating significant differences in sleep quality,degree of drowsiness,and memory scores before and after treatment.A statistical analysis was also conducted on the SAS,HAMA,SDS,and HAMD scores of sleep deprivation subjects before and after treatment.After treatment,the SAS,HAMA,SDS,and HAMD scores of the sleep deprivation subjects decreased.There were statistically significant differences in SAS(p<0.01),HAMA(p<0.05),and SDS,HAMD scores(p<0.01),indicating significant differences in anxiety and depression scores before and after treatment.4 The related results of "Adjust Zang and Arouse Spirit" electroacupuncture intervention on sleep deprivation model rats:(1)Body weight: Week 0(before sleep deprivation): There was no statistically significant difference in body weight among rats in each group(p>0.05).Week 1(1 week of sleep deprivation): Compared with the Con group,the body weight of rats in the other groups decreased,with statistically significant differences(p<0.01).Compared with the Mod group,there was no statistically significant difference in the decrease of body weight in the West and Acup groups(p>0.05).Week 2(2 weeks of sleep deprivation): Compared with the Con group,the decrease in body weight of rats in the Mod group was statistically significant(p<0.01).Compared with the Mod group,there was no statistically significant difference in the decrease of body weight in the West and Acup groups(p>0.05).Week 3(1week of intervention treatment): Compared with the Con group,the decrease in body weight of rats in the Mod group was statistically significant(p<0.01).Compared with the Mod group,there was no statistically significant difference in the decrease of body weight in the West group(p>0.05),while the body weight of rats in the Acup group increased with a statistically significant difference(p<0.05).Week 4(2 weeks of intervention treatment):Compared with the Con group,the decrease in body weight of rats in the Mod group was statistically significant(p<0.01).Compared with the Mod group,the body weight of rats in both the West and Acup groups increased with statistically significant differences(p<0.01).(2)Morris Water Maze: Comparison of Escape Latency: Compared with the Con group,the latency time of rats in the Mod group on Days 1-5 was significantly increased with a statistically significant difference(p<0.01).When comparing the West and Acup groups with the Mod group,there was no significant shortening of latency in the Day 1 West group(p>0.05),while there was a significant shortening of latency in the Acup group with a statistically significant difference(p<0.05).On Days 2-4,rats in the West group showed a shortened latency with a statistically significant difference(p<0.05),while rats in the Acup group showed a significant shortening of latency with a statistically significant difference(p<0.01).On Day 5,both the West and Acup groups showed a significant shortening of latency with a statistically significant difference(p<0.01).Comparison of Frequency of Crossing the Platform and Entering the Target Quadrant: Compared with the Con group,the frequency of rats in the Mod group crossing the platform and entering the target quadrant was significantly reduced(p<0.01),indicating a statistically significant difference.When comparing the West and Acup groups with the Mod group,the frequency of rats crossing the platform(both p<0.01)and entering the target quadrant(p<0.05 and p<0.01)increased significantly,indicating statistically significant differences.Comparison of Swimming Distance and Time: Compared with the Con group,the ratio of distance and time in the target quadrant decreased in the Mod group(p<0.01),indicating a statistically significant difference,while the ratio of distance and time in the sleep quadrant increased(p<0.01),also indicating a statistically significant difference.When comparing the West and Acup groups with the Mod group,there were no significant changes in the ratio of distance and time in the target and entry quadrants in the West group(p>0.05),indicating no statistically significant difference.However,in the Acup group,the ratio of distance and time in the target quadrant increased significantly(p<0.01),indicating a statistically significant difference,while the ratio of distance and time in the entry quadrant decreased significantly(p<0.01),also indicating a statistically significant difference.(3)Open Field Test: Compared to the Con group,the Mod group rats showed significant reductions in total path length,activity time,and center entries(p<0.01,p<0.01,p<0.05),with a significant increase in immobility time(p<0.01),indicating statistically significant differences.Although the time spent in the center by the Mod group rats did not reach statistical significance,there was a clear trend of decrease.When compared to the Mod group,the West group rats showed significantly increased total path length and activity time(p<0.05,p<0.01),with a significant decrease in immobility time(p<0.05).The Acup group rats exhibited significant increases in total path length,activity time,and center entries(p<0.01,p<0.05,p<0.05),with a significant decrease in immobility time(p<0.05),indicating statistically significant differences.(4)Circadian Clock Genes:Compared to the Con group,the expression levels of Clock,Bmal1,Cry1,and Per2 m RNA in the Mod group rats were significantly decreased,with statistically significant differences(p<0.01).When compared to the Mod group,the expression values of Clock,Bmal1,Cry1,and Per2 m RNA in the West group rats were significantly higher than those in the Mod group,with statistically significant differences(p<0.01,p<0.05,p<0.01,p<0.01).In the Acup group,the expression levels of Clock,Cry1,and Per2 m RNA were significantly higher than those in the Mod group,with statistically significant differences(p<0.01,p<0.05,p<0.01).However,there was no significant difference in the expression level of Bmal1 m RNA(p>0.05).Compared to the Con group,the expression levels of Clock,Bmal1,Cry1,and Per2 proteins in the Mod group rats were significantly decreased,with statistically significant differences(p<0.01).When compared to the Mod group,the expression values of Clock,Bmal1,Cry1,and Per2 proteins in the West group rats were significantly higher than those in the Mod group,with statistically significant differences(p<0.01,p<0.05,p<0.01,p<0.05).In the Acup group,the expression levels of Clock,Cry1,and Per2 proteins were significantly higher than those in the Mod group,with statistically significant differences(p<0.01,p<0.05,p<0.05).However,there was no significant difference in the expression level of Bmal1 protein(p>0.05).(5)Glu/GABA-Gln metabolic pathway: Compared to the Con group,the Glu level in the hypothalamus of rats in the Mod group was significantly increased(p<0.01),while the GABA level was significantly decreased(p<0.01),resulting in a significant increase in the Glu/GABA ratio(p<0.01).When compared to the Mod group,the Glu level in the hypothalamus of rats in the West group and Acup group was significantly reduced(p<0.01). The GABA level in the West group was significantly increased(p<0.05),and that in the Acup group was significantly increased(p<0.01).Consequently,the Glu/GABA ratio in both groups was significantly decreased(p<0.01).Compared to the Con group,the expression of GABA A receptor protein in rats in the Mod group was significantly increased,and the difference was statistically significant(p<0.05).When compared to the Mod group,the expression of GABA A receptor protein in rats in the West and Acup groups was significantly lower than that in the Mod group,and the difference was statistically significant(p<0.05).Compared to the Con group,the expression of GABAARα1 in rats in the Mod group was significantly increased,and the difference was statistically significant(p<0.01).When compared to the Mod group,the expression of GABAARα1 in rats in the West and Acup groups was significantly lower than that in the Mod group,and the difference was statistically significant(p<0.01).Conclusion:1 Rooted in Professor Wang Hongfeng’s clinical expertise in acupoints for sleep deprivation intervention,this study aims to refine the "Adjust Zang and Arouse Spirit" acupuncture method by harnessing cutting-edge data mining techniques.This endeavor aims to provide a robust scientific foundation for enhancing the clinical efficacy of the treatment.2 The application of "Adjust Zang and Arouse Spirit" electroacupuncture significantly enhances sleep quality among individuals experiencing sleep deprivation.It effectively alleviates symptoms such as daytime sleepiness,memory decline,anxiety,and depressive moods,thereby improving their overall well-being.3 Not only does the "Adjust Zang and Arouse Spirit" electroacupuncture effectively mitigate weight loss trends in rats with sleep deprivation,but it also helps alleviate their anxious and depressive behaviors,leading to a more balanced physiological state.4 By modulating key circadian clock genes such as Clock,Cry1,and Per2,as well as the Glu/GABA-Gln metabolic pathway,the "Adjust Zang and Arouse Spirit" electroacupuncture aims to rectify circadian rhythm disturbances induced by sleep deprivation.This approach helps to improve cognitive abilities such as learning and memory,while also mitigating symptoms of anxiety and depression. |
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