Clinical Observation And Related Mechanism Research Of Herb-Partitioned Moxibustion On The Navel In The Treatment Of Insomnia With Deficiency Of Both Heart And Spleen

Objective:1.To observe the clinical efficacy of herb-partitioned moxibustion on the navel in the treatment of insommia with deficiency of both heart and spleen.2.To observe the effects of herb-partitioned moxibustion on the navel on levels of hippocampal glutamate（Glu）,γ-aminobutyric acid（GABA）,serotonin（5-HT）and serotonin-1A receptor(5-HT_1AR)in experimental insomnia rats,and to explore the effect mechanism of herb-partitioned moxibustion on the navel on insomnia from perspective of Glu/GABA-Gln metabolic loop,and regulation of 5-HT and 5-HT_1A receptor system.Methods:Clinical research:In this study,a prospective,layered,randomized,single-blind,parallel controlled clinical trial design method was adapted.84 insomnia patients with deficiency of heart and spleen syndrome who met the inclusion criteria were selected and divided into treatment group（42 cases）and control group（42 cases）by stratified randomization.The treatment group was treated with herb-partitioned moxibustion on the navel,while the control group was treated with starch-partitioned moxibustion on the navel.The treatment was performed every 5 days for a total of 6 treatments.The curative effect comparison was analyzed by Insomnia Severity Index（ISI）,Pittsburgh Sleep Quality Index（PSQI）,TCM Syndrome Scale and Fatigue Scale-14（FS-14）before and after treatment.The scores of PSQI and ISI were regarded as primary indicators for therapeutic effect,and the scores of FS-14 and TCM Syndrome Scale were considered as secondary indicators for therapeutic effect.Experimental study:48 SPF male SD rats were randomly divided into normal control group,model control group,herb-partitioned moxibustion group and starch-partitioned moxibustion group,with12 cases in each group.The model of insomnia was established by PCPA intraperitoneal injection.After model establishment,rats in the herb-partitoned moxibustion group and the starch-partitioned moxibustion group were treated with herb-partitioned moxibustion and starch-partitioned moxibustion on the navel.Rats in the normal control group and the model control group were subjected to the same grasping and fixation stimulation,but no treatment was given.The treatment was performed once daily for 8 consecutive days.On the 12^th day of modeling,the open field test（OFT）was performed,and the hippocampal tissue was collected.Enzyme linked immunosorbent assay（ELISA）and ultraviolet-visible spectrophotometry（UV-Vis）were applied to detect the contents of Glu,GABA and 5-HT in hippocampal tissue,and the positive expression of 5-HT_1AR mRNA in hippocampus was detected by Real-time PCR（q PCR）.Results:Clinical research:1.In this clinical trial,a total of 84 insomnia patients with deficiency of heart and spleen syndrome who met the inclusion criteria were observed,including 3 cases of detachment,and finally 40 cases in the herb-partitioned moxibustion group and 41 cases in the starch-partitioned moxibustion group.There were no statistically significant differences between the treatment group and the control group in terms of PSQI,ISI,TCM syndrome scale and FS-14 scores at baseline（P>0.05）,and the general data of the two groups were balanced and comparable（P>0.05）.2.The influence of herb-partitioned moxibustion on the navel on PSQI in insomnia patients with deficiency of both heart and spleen.Compared with pre-treatment,the total scores of PSQI scale in the treatment group and control group were significantly decreased（P<0.01）.The total score of PSQI scale in the treatment group was significantly lower than that of the control group after treatment（P<0.01）.3.The influence of herb-partitioned moxibustion on the navel on ISI in insomnia patients with deficiency of both heart and spleen.After treatment,the total scores of ISI in the treatment group and the control group were decreased significantly compared with those before treatment（P<0.01）.Compared with the control group,the total score of ISI in the treatment group was lower with statistical significance after treatment（P<0.05）.4.The influence of herb-partitioned moxibustion on the navel on TCM syndromes in insomnia patients with deficiency of both heart and spleen.After treatment,the total scores of TCM syndromes in the treatment group and the control group were significantly lower than before treatment（P<0.01）.After treatment,the total score of TCM syndromes in the treatment group was lower than that in the control group with statistical significance（P<0.05）.5.The effect of herb-partitioned moxibustion on the navel on FS-14 in insomnia patients with deficiency of both heart and spleen.After treatment,the total scores of FS-14 in the treatment group and the control group were decreased significantly compared with those before treatment（P<0.01 or P<0.05）.Compared with the control group,the total score of FS-14 in the treatment group was lower with statistical significance after treatment（P<0.05）.Animal experiment:1.The effect of PCPA modeling on the sleep latency and sleep duration in the pentobarbital sodium-induced sleep tests.The results of PCPA modeling in the pentobarbital sodium-induced sleep tests showed that the sleep latency of the three groups of model rats was significantly longer than that of the normal control group（P<0.01）,and the sleep duration of the three groups of model rats was significantly shorter than that of the normal control group（P<0.01）.There was no significant difference in sleep latency and sleep duration among the three groups of modeled rats（P>0.05）.2.The effect of herb-partitioned moxibustion on the navel on the standing times,resting time,total distance of exercise and number of fecal pellets in experimental rats in the open field test.Compared with the blank control group,the resting time in the model control group was significantly increased（P<0.01）,and the standing times,total distance of exercise and number of fecal pellets in the model control group were significantly decreased（P<0.01）.The resting time in the herb-partitioned moxibustion group was significantly increased compared with the model control group（P<0.01）,while the standing times,number of fecal pellets and total distance of exercise in the herb-partitioned moxibustion group were significantly decreased compared with the model control group（P<0.01）.The number of fecal pellets in the starch-partitioned moxibustion group was significantly decreased compared with the model control group（P<0.05）,and there was no significant difference in the standing times,total distance of exercise and resting time compared with the model control group（P>0.05）.The resting time of the herb-partitioned moxibustion group was significantly longer than that of the starch-partitioned moxibustion group（P<0.01）,while the total distance of exercise and standing times of the herb-partitioned moxibustion group were significantly less than those of the starch-partitioned moxibustion group（P<0.01）.There was no statistically significant difference in the number of fecal pellets between the herb-partitioned moxibustion group and the starch-partitioned moxibustion group（P>0.05）.3.The effect of herb-partitioned moxibustion on the navel on the content of Glu and GABA in the hippocampus of insomnia rats.The hippocampal Glu content of the model control group was significantly higher than that of the normal control group,while the hippocampal GABA content of the model control group was significantly lower than that of the normal control group（P<0.01）.The content of Glu in the hippocampus of the herb-partitioned moxibustion group was significantly lower than that of the model control group,while the content of GABA in the hippocampus of the herb-partitioned moxibustion group was significantly higher than that of the model control group（P<0.01）.The hippocampal GABA content of the herb-partitioned moxibustion group was significantly higher than that of the starch-partitioned moxibustion group（P<0.05）,and there was no statistically significant difference in the content of Glu in the hippocampus between two groups（P>0.05）.4.The effect of herb-partitioned moxibustion on the navel on the 5-HT content and the 5-HT_1AR mRNA expression levels in the hippocampus of insomnia rats.Compared with the normal control group,the content of 5-HT and the relative expression of 5-HT_1AR mRNA in the hippocampus of the model control group were lower with statistical significance（P<0.01）.The content of 5-HT and the relative expression of5-HT_1AR mRNA in the hippocampus of the herb-partitioned moxibustion group and the starch-partitioned moxibustion group were significantly higher than those of the model control group（P<0.01 or P<0.05）.The content of 5-HT and the relative expression of5-HT_1AR mRNA in the hippocampus of the herb-partitioned moxibustion group were significantly higher than those of the starch-partitioned moxibustion group（P<0.05）.Conclusions:1.The herb-partitioned moxibustion on the navel can effectively improve sleep condition,fatigue state and TCM symptoms in mild and moderate insomnia patients with deficiency of both heart and spleen.Herb-partitioned moxibustion on the navel is more effective than starch-partitioned moxibustion on the navel in improving sleep condition,fatigue state and TCM symptoms for mild and moderate insomnia patients with deficiency of both heart and spleen.2.The treatment of herb-partitioned moxibustion on the navel can significantly reduce the influence of insomnia on the spontaneous activities of PCPA-treated rats,effectively regulate the standing times,resting time,total distance of exercise and number of fecal pellets in the open field experiment of insomnia rats during the daytime,and relieve the anxiety,tension and excitability.Compared with starch-partitioned moxibustion on the navel,herb-partitioned moxibustion on the navel is more effective in improving spontaneous activities and exploratory behaviors of PCPA-treated rats in new surroundings in the daytime.3.The treatment of herb-partitioned moxibustion on the navel can improve the levels of GABA,5-HT and 5-HT_1AR mRNA in the hippocampus of PCPA-treated rats,and reduce the level of hippocampal Glu in insomnia rats.The treatment of herb-partitioned moxibustion on the navel has certain curative effect on the regulation of sleep-wake cycle in insomnia rats,and its mechanism is possibly related with regulating the unbalanced Glu/GABA-Gln metabolic loop and controlling the levels of 5-HT and 5-HT_1A receptor in the hippocampus of insomnia rats.Compared with starch-partitioned moxibustion on the navel,herb-partitioned moxibustion on the navel is more effective in the regulation of Glu/GABA-Gln metabolic loop and the expression of 5-HT and 5-HT_1A receptor in the hippocampus of insomnia rats.