Acupuncture Regulates The Influence Of DEC1 And DEC2 Gene Expression In The Liver Of Peripheral Tissues

Objective: To be observed the effects of proteins expression,which acupuncture BL18 on the expression of DEC1 mRNA,DEC2 mRNA,DEC1,and DEC2 in mice with disturbed sleep-wake cycle,research the acupuncture BL18 regularion function on liver-sleep arousal cycle.Methods: 72 C57BL/6J mice were randomly divided into blank group,model group,acupuncture BL18 group,non-acupoint group with a total of four groups of 18 in each group.In the Light: 08:00-20:00,Dark: 20:00-08:00 in the condition of the switch lamp for 7 days.The 8th day of the experiment that mice finished the acclimation,,using the short-term deprivation protocol 2h/2h light stimulation modeling the Sleep-acoustic Cycle Disorder Model was obtained by the method,and the model was established for 5 days.After confirming the success of the modeling,the intervention was performed on the 13 th day of the experiment: the blank group was not treated,and the model group was bundled.The acupuncture group and non-acupoints were given the electro-acupuncture intervention 5 minutes at 15:00 for 5 consecutive days.The BL18 group was acupuntured by filiform needles inner and downward obliquely 0.4 cm on the both BL18 with the electroacupuncture which is the 6805-AII elecroacupunture instrument.The electroacupuncture stimulation parameters were: density wave was 4/20Hz;density cycle was 6 seconds,and the current was adjusted to the tail or the lower extremity slightly twitches.Non-acupoint group selected mice double and fixed under stress,and the acupuncture method was the same as acupuncture in the BL18 group.At 2 a.m.on the 18 th day of the experiment(12 hours after the end of the acupuncture),3 mice were sacrificed every 4 hours for liver tissue.The whole processes were monitored with a 360 smart camera and were recorded datas of spontaneous standing and activities.Combining with the Real-Time PCR,Western Blot method be observed in the liver DEC1 mRNA,DEC2mRNA rhythmic expression and affects the quantity of DEC1,DEC2 expression,and then rearched the regulation function of the effect of Acupuncture in BL18 on Sleep-Wakefulness Cycle.Result: 1.All groups of C57BL/6J mice showed irritability,high excitability,strong offensive behavior and other obvious behavioral changes after building model.The number of standing and spontaneous activities was significantly increased,and the weight gain was inhibited.The results were compared with those before modeling.There was statistical significance(P<0.05);after 5 days of electroacupuncture,the behavioral improvement of C57BL/6J mice in the BL18 group significantly decreased the number of standing and spontaneous activities,and the weight recovery increased,compared with the model group and without The non-acupoint group had statistical significance(P<0.05),and there was no statistical significance compared with the blank group(P>0.05).2.In addition to the blank group,the DEC1 mRNA and DEC2 mRNA in the liver of each group showed obvious rhythm disorder,P<0.3.However,the expression levels of DEC1 and DEC2 proteins were significantly decreased,compared with pre-modeling,P<0.05,which was statistically significant.3.After electroacupuncture at BL18 group for 5 days,compared with the model group,liver RT-PCR in the acupuncture BL18 group showed that DEC1 mRNA and DEC 2 mRNA recovered their autonomous rhythm,P < 0.3;while non-acupoint group still showed rhythm Disorder,P>0.3.4.After electroacupuncture BL18,the expression of DEC1 and DEC2 protein in the BL18 group increased.Compared with the model group and non-acupoint group,there was statistical significance(P<0.05);There was no statistical difference between the group and model group(P>0.05).CONCLUSION: 1 Electroacupuncture at BL18 has a clear regulation effect on C57BL/6J mice with disturbed sleep-wake cycle,and can restore C57BL/6J mice whose sleep-wake cycle that is disordered to their autonomous rhythm.2 Electroacupuncture at BL18 can restore the rhythm of the negative feedback regulatory genes DEC1 and DEC2,and upregulate the expression of DEC1 and DEC2 proteins.The mechanism may be to regulate the sleep-wake cycle by reducing the awakening time and increasing the sleep time.