Study On Mechanism Of Clinical Effects Of "Ni Zhen" Method On Sleep Deprivation And Immune Protein Regulation

Objective: To evaluate the clinical effects of "Ni Zhen" method on healthy male youths after 24-h total sleep deprivation,and to observe its effects on emotional experience,fatigue and drowsiness during 24 h total sleep deprivation,as well as its results on the alteration of immune-related proteins and related pathways in human plasma samples.The mechanism of the biological effect of acupuncture in the prophylactic treatment of sleep deprivation.Methods:Clinical trial studies1.Healthy subjects baseline period assessment.The TPF health sleep intelligence management system was used to screen healthy subjects who met the requirements,and the differences in age,body temperature,height,body mass,BMI,PSQI score,systolic blood pressure,diastolic blood pressure,respiration,resting heart rate,SAS,SDS score,daily sleep time and TPF sleep score between the pre-acupuncture acupuncture point group,non-acupuncture point group and blank group were observed and compared during the baseline period of the experiment.2.Study of the effect of the "Ni Zhen" method on the 24 h brain neurochemical transmitters in TSD.The ET brain function analyzer was used to observe the differences of brain neurochemical transmitters GABA,Glu,5-HT,Ach,NE and DA in the pre-acupuncture point group,non-acupuncture point group and blank group at different time points compared with the normal value range.3.Study on the effect of "Ni Zhen" method on the fatigue and drowsiness of TSD24 h.The ET brain function analyzer was used to observe the differences in brain fatigue,brain ischemia and hypoxia,and brain excitation and inhibition in the pre-acupuncture point group,non-acupuncture point group and blank group at different time points.The differences in the prognosis of the three groups were observed in the FSS scale score,BFI scale score,and SSS scale score of the subjects at several different time points.4.Study of the effect of the "Ni Zhen" method on the mood of TSD 24 h.The ET brain function analyzer was used to observe the difference of subjects' ability to control emotion at different time points in the pre-acupuncture point group,non-acupuncture point group and blank group.The Face Reader facial expression analysis system was used to observe the differences in the basic human emotion values,PANAS scale positive emotion and negative emotion score values among the three groups at several different time points.Effect mechanism studyBased on the "4D Label-free" proteomics technology,we screened plasma samples of human peripheral blood for differential proteins and enriched the KEGG database and Inter Pro database for their molecular functions and protein pathways.We compared the immune-related differential proteins and enrichment pathways in each group,and explored the material basis and molecular mechanism of the immunomodulatory effect of the "Ni Zhen" method.Results:Clinical trial studies1.Baseline period assessment in healthy subjects.There was no statistical difference in the data values of age,height,body mass,body temperature,BMI,respiration,resting heart rate,and blood pressure between the three groups of subjects at baseline(P>0.05).In terms of emotional state,there was no statistical difference between the three groups in terms of SAS score and SDS score in two comparisons(P>0.05);in terms of sleep duration and quality,there was no statistical difference between the three groups in terms of PSQI score,deep,moderate and light sleep duration in two comparisons(P>0.05).2.Study on the effect of "Ni Zhen" method on brain neurochemical transmitters at 24 h of TSD.After 24 h of TSD,the GABA values in the blank group showed an increase compared to the baseline and normal values,with statistical differences(P<0.05).24 h of TSD,the acupoint group showed a decrease compared to the non-acupoint group(P<0.05).The cave group showed a decrease compared with the blank group(P<0.05).For the comparison of the normative values,the values of Glu,5-HT,Ach,NE and DA contents in the three groups were not statistically significant compared with the normal values(P>0.05),and there were no statistical differences between the three groups when compared with each other(P>0.05).3.Study on the effect of "Ni Zhen" method on the fatigue and drowsiness of TSD24 h.After 24 h of TSD,the ischemic and hypoxic values of the brain and fatigue values of the blank group showed an increase compared with the baseline values(P<0.05),and the excitation and inhibition values of the brain showed a decrease(P<0.05).The ischemic-hypoxic and fatigue values of the brain in the acupuncture group showed a decrease(P<0.05)and the excitatory-inhibitory values of the brain in the blank group showed an increase(P<0.05)compared with the baseline values.after 24 h of TSD,the SSS,FSS,and SSS scale scores in the blank group showed an increase(P<0.05)compared with the baseline values.The BFI and FSS scale scores in the acupuncture group showed a decrease compared to the blank group(P<0.05).The FSS score values in the acupuncture point group showed a decrease compared to the non-acupuncture point group(P<0.05).4.Study of the effect of "Ni Zhen" method on the mood of TSD at 24 h.Emotional control ability: After 24 h of TSD,the blank group showed a decrease in emotional control ability compared to the baseline value(P<0.05).The acupuncture point group showed an increase compared to the blank group(P<0.05).The acupuncture point group showed an increase compared to the non-acupuncture point group(P<0.05).As for emotion detection: after 24 h of TSD,there was no statistical difference between the three groups in two comparisons(P>0.05).As for positive and negative emotion values: after 24 h of TSD,the PANAS positive emotion values of the blank group showed a decrease compared to the baseline values(P<0.05),and the negative emotion scores showed an increase compared to the baseline values(P<0.05).For positive emotions: the acupuncture point group showed an increase compared to the blank group(P<0.05).The acupuncture point group showed an increase compared to the non-acupuncture point group(P<0.05).Negative emotions: the acupuncture point group showed a decrease compared to the blank group(P<0.05).The acupuncture point group showed a decrease compared to the non-acupuncture point group(P<0.05).Mechanism of effect studyA total of 1563 quantifiable proteins were identified.before and after TSD24 h,the number of immune-related proteins was compared: 30 in the blank group,36 in the non-acupuncture point group,and 45 in the acupuncture point group.There were 36 proteins that differed in the cave group compared to the blank group,31 proteins that differed in the non-cave group compared to the blank group,and 9 proteins that differed in the non-cave group compared to the cave group.Nine key proteins were enriched.Differential proteins were enriched to 8 pathways before and after TSD24 h in the acupuncture point group,1pathway in the blank group,and 2 pathways in the non-acupuncture point group.All p<0.05 for the above comparisons.Conclusion:1.TSD24 h can have important effects on human fatigue,drowsiness and emotions,mainly manifested as increased human brain fatigue,increased brain ischemia and hypoxia,increased brain inhibition,weakened emotional control,reduced positive emotions and increased negative emotions.2.The "Ni Zhen" method has a positive regulating effect on the value of GABA content in human brain after 24 h of TSD,and can delay the trend of increasing GABA content in human brain after 24 h of TSD,and the effect of acupuncture points is better than non-acupuncture points.3.The "Ni Zhen" method has a moderating effect on the adverse effects of increased brain fatigue,increased brain ischemia and hypoxia,increased brain inhibition,decreased emotional control,decreased positive emotions and increased negative emotions triggered by TSD24 h,and can effectively reduce their occurrence,and the effect of acupuncture points is better than that of non-acupuncture points.4.From the proteomic analysis,it was demonstrated that Fatty acid biosynthesis,Platelet activation,ABC transporters pathway can be changed after 24 h of TSD in healthy human.The "Ni Zhen" method can cause changes in immune-related differential protein content and protein pathways such as AMPK signaling pathway and Toll-like receptor signaling pathway,and these proteins and protein pathways may be responsible for the changes in the immune system.These proteins and protein pathways may be the key proteins and key pathways for the effect of "reverse needle method".