A Randomized Controlled Study On Acupuncture Treatment Of "Siguan" Points For Depression And The AC-cAMP-PKA Postreceptor Signal Transduction Mechanism

Objectives1.From a randomized, single blind and controlled trial, we explore the effectiveness and safety of acupuncture treatment of "Siguan" acupoints on depression of stagnation of liver-Qi type.2. To explore the mechanism of acupuncture for antidepressive effect on AC-cAMP-PKA postreceptor signal transduction pathway.Methods1. Clinical study108cases were randomized into the following groups:Siguan acupoints group, non-acupoint group and Streitberger needle group groug.Siguan acupoints group:Siguan points (bilateral Hegu (LI4) and Taichong(LR3)) were selected to received acupuncture. After sterilizing,0.35×40mm acupuncture needles were put through the needle pipe and vertically inserted below to the skin through the foam tray which was stick to the surface of the points. Manipulated the needle until de-qi was felt by the patient. Retained the needle for30min with needle manipulation once per15min.Non-acupoint group:Two points that correspond to Siguan points were located①on the dorsum of hand and between the middle of the3th and4th metacarpal bone;②on the3th lateral metatarsal bone on dorsum pedis, between the3th and4th metatarsal bone. Needle manipulation and retaining were similar to Siguan points group.Streitberger needle group:The selected points and the needle insertion procedure were similar as Siguan acupoints group, except that the tip of the needle was blunt and could not puncture into the skin. During the needle retaining, the needle was hold by the foam tray and the needle pipe and adjust the stretching pipe to keep the appearance of the placebo needle look like as inserted into the skin as the verum needle in Siguan points group.All the patients in the three groups were asked to deep breath to regulating qi during needle retaining and received3sessions weekly with total6weeks of treatment.The effects were evaluated by the HAMD Scales, the SDS Scales, and the improvement Scales of symptoms of Traditional Chinese Medicine before the treatment and4time points (at the end of2week,4week and6week) during the treatment course. ITT analysis were used for all the data analysis.2. Animal studyAdult Sprague-Dawley male rats (200-250g) were used in the present experiment. Animals were divided randomly into7groups:the control group, the model group, the electroacupuncture(EA) group and drug group, EA plus normal saline (NS)group, EA plus H89group and Fluo plus H89group;8rats in each group. The chronic mild stress model(CMS) of depression was induced by various kinds of stress for42days. In the model group, rats were exposed to chronic mild unpredictable mild stress for6weeks. In the EA group, rats receive CMS procedure and electroacupuncture stimulation at Siguan points (bilateral Hegu (LI4) and Taichong(LR3)) once every other day for3weeks after CMS exposure. In the drug group, rats receive the same treatment as the CMS group and Fluoxetine treatment everyday for3weeks after CMS. The groups plus H89/NS were conditioned with the same as the EA or Fluoxetine group except for the intracerebroventricular injection of H89between CMS and EA. Changes of open-field behavior, sucrose consumption, AC activity, cAMP level and PKA activity in hippocampus of rats were observed in the study.Result1.Clinical study1.1BaselineThe difference in terms of age, height, weight, disease duration, sex, educational level, occupation, living habits, history of western medicine taking or acupuncture treatment receiving among3groups is no significant (P>0.05); And, the difference of scores of HAMD scale, SDS scale and TCM syndrome rating scale among the three groups of depressed patients on baseline was not statistically significant(P>0.05), baseline level can be considered the same among the three groups and comparability. 1.2Total effective rate and excellence rateAfter6weeks of treatment, the total effective rate and the excellence rate of the3groups respectively were84.21%and44.74%in Siguan acupoints group,67.65%and14.71%in non-acupoint group,38.89%and5.56%in the Streitberger needle group. Compare the total effective rate of the3groups, there was a significant difference among the3groups (P<0.0167).1.3HAMD ScaleThe gross effective rate was higher and the HAMD Scale score was better in the Siguan acupoints group than that of the non-acupoint group and sham-acupuncture group (P<.05). The improvement of score of anxiety/somatization factor, body weight factor, cognitive handicap factor, blockage factor and the sleeping disorder factor in Siguan acupoints group was started on the2week during the treatment, and which was lasted to the end of the treatment.1.4SDS scaleThe SDS Score was better in the both acupuncture groups than that in the non-acupoint group and Streitberger needle group (P<0.0167, P<0.0167), however, no significant difference was found between the non-acupoint group and Streitberger needle group.1.5Improvement Scales of symptoms of Traditional Chinese MedicineThe Improvement Scales of symptoms of TCM Score was better in the both acupuncture groups than that in the non-acupoint group and Streitberger needle group (P<0.0167,P<0.0167), again, no significant difference between the non-acupoint group and Streitberger needle group.2. Animal study2.1Changes of openfield behaviorThe behavior (horizontal and vertical motion) of the depression rats decreased significantly following stress, and EA treatment as well as EA+NS can effectively improve it, which is better than that of the drug (P<0.05, P<.0.05, P<0.05). Enhanced effects induced by EA were reversed by H89pretreatment.2.2Changes of sucrose consumptionSucrose intake in all stress groups decreased significantly in the3rd week and was much less than the control group (P<0.05). However, there was no remarkable difference among all stress groups. EA or Fluoxetine treatment leaded to increase in sucrose intake in the sixth week, however, not indicating significant difference with CMS group. H89pretreatment inhibited obviously increase in sucrose intake induced by EA.2.3AC activity, cAMP level and PKA activityAnimals exposed to stressors also showed a significant decrease in the ratio of AC transformation after the last week when the experiment ended. In the EA group and Fluxitine group, the ratio of AC transformation increased significantly after EA treatment for3weeks compared to the CMS group. EA and Fluxitine can improved the level of AC transformation ratio in the hippocampus of rats to a level as of normal rats. Changes in cAMP level and PKA activity were similar to AC (P<0.05, P<0.05, P<0.05)Conclusion1. Acupuncture treatment of Siguan acupoints was effective and safe in treating depression.2. Acupuncture treatment of Siguan acupoints regulating liver showed a more obviously curative effect on all factors of depression after treatment when compared to non-acupoint group and Streitberger needle groug.3. Acupuncture could improve signif ieantly changes of the behavior, sucrose consumption, AC activity, cAMP level and PKA activity in hippocamp of rats, therefore, AC-cAMP-PKA postreceptor signal transduction pathway is an important approach and target of acupuncture in treating depression.