Study On Acupuncture Regulating The Function Of TRN Neurons In Rats With Sleep Wake Cycle Disorder

Objective:To explore the mechanism of acupuncture regulating sleep-wake cycle disorder in PCPA insomnia rats by taking the thalamic regulating nucleus(TRN)as the core,the discharge of neurons and the number of PV(Parvalbumin)positive neurons in TRN area as the breakthrough point.In order to provide experimental basis for clinical treatment of sleep disorder caused by sleep-wake cycle disorder.Method:SPF male SD rats were domesticated according to the normal light and dark cycle(L:D=12:12)for at least one week,and 30 rats synchronized with the normal light and dark cycle were randomly divided into control group(n=10)and model group(n=20).The insomnia model was induced by intraperitoneal injection of Chlorophenylalanine(PCPA)suspension for 3 days.According to the activity during the light period,the rats were randomly divided into model group(n=10)and acupuncture group(n=10).After successful modeling,the acupuncture group selected Neiguan(PC6)and Zusanli(ST36)at the time point of ZT4(ZT:Zeitgebertime,11:00)for continuous acupuncture for 5 days,once a day,15 minutes each time.Clocklab3 data acquisition system was used to dynamically monitor the sleep condition of rats throughout the experiment.The electrophysiological technique was used to collect and analyze the electrical activity of TRN neurons in PCPA insomnia rats,and the immunofluorescence labeling technique was used to analyze the changes of PV positive neurons in TRN area of PCPA insomnia rats.Result:1.The rhythm of spontaneous wheel activity of SD rats in each group had similar circadian rhythm during domestication.There was no difference in peak phase of spontaneous wheel activity in each group(P>0.05),and the baseline was the same.PCPA rats showed yellow hair,increased hair loss,sensitive to grasping reaction,restlessness or aggression.The spontaneous wheel activity of rats in the acupuncture group and the model group increased significantly during the light rest period,and the peak phase showed advanced transfer(P<0.05),indicating that the circadian rhythm of spontaneous activity was disordered and the model was established successfully.After acupuncture treatment,the peak phase of wheel spontaneous activity in the acupuncture group recovered from the light rest period to the dark phase,which was significantly different from that in the model group(P<0.05).2.Comparing the discharge frequency and pattern of TRN neurons in each group during the light rest period,it was found that the discharge frequency in the model group was lower than that in the control group(P<0.05),showing a mixed pattern(single regular discharge and regular discharge),while the discharge frequency in the acupuncture group was higher than that in the model group(P<0.05),and the discharge mode was mainly cluster discharge.3.The immunofluorescence staining data of the three groups were analyzed.Compared with the control group,the average fluorescence density of the model group decreased(P<0.05),the number of PV positive neurons decreased significantly,and the fluorescence color was dim.Compared with the model group,the average fluorescence density and the number of PV positive neurons in the acupuncture group increased(P<0.05),suggesting that acupuncture can increase the number of PV positive neurons.Conclusion:1.Acupuncture at PC6 and ST36 can effectively adjust the disorder of sleep-wake cycle after sleep disorder.2.Acupuncture at PC6 and ST36 regulates the disorder of sleep-wake cycle,which may be related to the regulation of the number of PV positive GABA neurons in TRN area and the improvement of neuronal excitability.