| [Background]Post-traumatic stress disorder(PTSD)is a long-term mental illness that occurs after a major traumatic event.Patients are often accompanied with depression,anxiety,sleep disorders and other symptoms,serious suicidal tendencies.This disease has the characteristics of high incidence rate,long course of disease and difficult to cure.In recent years,with the continuous emergence of major infectious diseases,wars and emergencies,the prevalence of PTSD showed a significant upward trend.PTSD has become a common disease that endangers human beings and causes serious burden to society.At present,sertraline and paroxetine are the main western drugs approved by the US Food and Drug Administration for the treatment of PTSD.However,long-term use of sertraline and paroxetine has inevitable side effects.Acupuncture treatment of PTSD has good curative effect,can significantly improve the symptoms of PTSD patients,such as mood,memory disorders and behavioral abnormalities,with little side effects.However,the mechanism of acupuncture in the treatment of PTSD is still unclear,which needs further study.In recent years,more and more attention has been paid to the role of apoptosis in the pathogenesis of PTSD.Endoplasmic reticulum stress-induced apoptosis is closely related to the occurrence and development of PTSD.Endoplasmic reticulum stress is caused by the accumulation of unfolded or misfolded proteins in endoplasmic reticulum.Persistent or excessive endoplasmic reticulum stress can activate unfolded protein response,which will eventually initiate apoptosis pathway and lead to neuronal apoptosis.The unfolded protein reaction is mainly composed of a molecular chaperone glucose regulatory protein 78(GRP78)and three transmembrane protein mediated signaling pathways.When long-term or severe stress occurs,GRP78 expression is up-regulated,which activates endoplasmic reticulum stress-specific apoptotic transcription factor C/EBP homologous protein(CHOP),and then activates Caspase-12 and Caspase-3,which eventually leads to cell apoptosis and participates in the pathogenesis of PTSD.However,the effect of acupuncture on endoplasmic reticulum stress related molecules in PTSD brain is still unclear.[Objective]In this study,the PTSD rat model was established by single prolonged stress(SPS),and the effects of acupuncture on the behavior of PTSD rats and the expression of GRP78,CHOP,Caspase-12 and Caspase-3 in hippocampus endoplasmic reticulum of rats were observed,so as to explore the possible mechanism of acupuncture treatment of PTSD and provide scientific basis for clinical treatment of PTSD.[Methods]1.Animal grouping:28 male SPF SD rats weighing(200 ± 20)g were randomly divided into normal group,model group,acupuncture group and sertraline group,with 7 rats in each group.2.Model preparation methods:PTSD rat model was established by the internationally recognized method of SPS.First,the rats were fixed and bound for 2 hours,then forced to swim for 20 minutes immediately(the rats were placed in a 50 cm high,20 cm diameter plexiglass tank with water depth of 30 cm and water temperature of 24℃);after drying and resting for 15 minutes,the rats were anesthetized with ether until they lost consciousness,and then they were taken out and put back into the cage after waking up.3.Intervention methodsMethods of acupuncture:from the first day after modeling,Baihui and Dazhui points were selected for treatment,10 minutes each time,once a day,a total of 7 times.The rats kept a free posture while retaining the needle.Administration method:sertraline(10mg/kg)was given by gavage from the first day after modeling,once a day,a total of 7 times.4.Behavioral observation:(1)Open field experiment:At the end of the experiment,the rats were put into the self-made open field box,and the times of climbing and standing,the time of entering the central area for the first time,the time of staying in the central area and the total distance of movement were recorded within 5 minutes to evaluate the autonomous activity and anxiety like behavior of rats.(2)Novelty inhibition feeding test:At the end of the experiment,rats were fasted for 24 hours,and could not help drinking water.The self-made box was paved with bedding material,and 8 pieces of feed with the same size were placed in the center of the box,and the shape of a ring was formed.The rats were put into the box at a fixed position,and the latency of the first feeding was recorded within 5 minutes.It was used to evaluate the anxiety like behavior of rats.(3)New object recognition experiment:At the end of the experiment,the exploration time(T new,T old)of new and old objects within 5 minutes of each rat was recorded,and the recognition ability of new objects was expressed by recognition index(%)=T new/(T New+T old)× 100.It was used to evaluate the cognitive function of rats.5.Index detection:(1)The expression of GRP78 mRNA and CHOP mRNA in rat hippocampus was detected by real-time fluorescent quantitative PCR(RT-PCR).(2)Western blot was used to detect the protein expression of Caspase-12 and Caspase-3 in rat hippocampus.6.Statistical methods:The experimental results are based on the mean ± Standard deviation(X ± s).The statistical software SPSS 20.0 was used and one-way ANOVA was used.Then LSD method was used to analyze the differences between groups.P<0.05 showed that the difference was significant.[Results]1.The effect of acupuncture on behavior of rats:(1)Effect of acupuncture on open field experiment of rats:Compared with the normal group,the number of crawling and standing times of the model group were significantly reduced(P<0.05).Compared with the model group,the number of rats in the acupuncture group increased significantly(P<0.05),and the number of standing times increased,but the difference was not statistically significant(P>0.05);The number of crawling and standing times of sertraline group were significantly increased(P<0.05).Compared with the acupuncture group,there was no significant difference in the number of crawling frames and standing times in the sertraline group(P>0.05).Compared with the normal group,the time of entering the central area for the first time in the model group increased significantly(P<0.01),the residence time in the central area decreased significantly(P<0.01),and the total distance of movement decreased significantly(P<0.05).Compared with the model group,the time of first entering the central area in the acupuncture group and sertraline group was significantly decreased(P<0.05,P<0.01),and the residence time and total distance of movement in the central area were significantly increased(P<0.01).Compared with the acupuncture group,there were no significant differences in the time of first entering the central area,the time of staying in the central area and the total distance of movement in the sertraline group(P>0.05).(2)Effect of acupuncture on novelty inhibition feeding experiment:Compared with the normal group,the feeding latency of the model group was significantly longer(P<0.05).Compared with the model group,the feeding latency of the acupuncture group decreased,but the difference was not statistically significant(P>0.05);The feeding latency of sertraline group was significantly decreased(P<0.05).Compared with the acupuncture group,there was no significant difference in the feeding latency of sertraline group(P>0.05).(3)Effect of acupuncture on new object recognition experiment in rats:Compared with the normal group,the new object recognition index of the model group was significantly lower(P<0.01).Compared with the model group,the new object recognition index of the acupuncture group was significantly higher(P<0.01);The new object recognition index of sertraline group increased,but the difference was not statistically significant(P>0.05).Compared with the acupuncture group,there was no significant difference in the recognition index of the sertraline group(P>0.05).2.Effect of acupuncture on the expression of GRP78 mRNA,CHOP mRNA,Caspase-12 and Caspase-3 in hippocampus of rats:(1)Effect of acupuncture on the expression of GRP78 mRNA in hippocampus of rats:Compared with the normal group,the expression of GRP78 mRNA in hippocampus of model group was significantly increased(P<0.05).Compared with the model group,the expression of GRP78 mRNA in hippocampus of the acupuncture group and sertraline group was significantly decreased(P<0.05).Compared with the acupuncture group,there was no significant difference in the expression of GRP78 mRNA in hippocampus of sertraline group(P>0.05).(2)Effect of acupuncture on the expression of CHOP mRNA in hippocampus of rats:Compared with the normal group,the expression of CHOP mRNA in the hippocampus of the model group was significantly increased(P<0.01).Compared with the model group,the expression of CHOP mRNA in hippocampus of the acupuncture group and sertraline group was significantly decreased(P<0.05).Compared with the acupuncture group,there was no significant difference in the expression of CHOP mRNA in the hippocampus of the sertraline group(P>0.05).(3)Effect of acupuncture on the expression of Caspase-12 protein in hippocampus of rats:Compared with the normal group,the expression of Caspase-12 protein in the hippocampus of the model group was significantly increased(P<0.05).Compared with the model group,the expression of Caspase-12 protein in hippocampus of the acupuncture group and sertraline group was significantly decreased(P<0.05).Compared with the acupuncture group,there was no significant difference in the expression of Caspase-12 protein in hippocampus of sertraline group(P>0.05).(4)Effect of acupuncture on the expression of Caspase-3 protein in hippocampus of rats:Compared with the normal group,the expression of Caspase-3 protein in hippocampus of model group was significantly increased(P<0.05).Compared with the model group,the expression of Caspase-3 protein in hippocampus of the acupuncture group decreased,but the difference was not statistically significant(P>0.05).Compared with the model group,the expression of Caspase-3 protein in hippocampus of sertraline group was significantly decreased(P<0.05).Compared with the acupuncture group,there was no significant difference in the expression of Caspase-3 protein in hippocampus of sertraline group(P>0.05).[Conclusion]1.The autonomic activity of PTSD rats decreased significantly,and there were anxiety like behavior and cognitive impairment.Acupuncture and sertraline can improve the anxiety like behavior of PTSD rats;Acupuncture can also improve the cognitive impairment of PTSD rats.2.The expressions of GRP78,CHOP and Caspase-12 in the hippocampus of PTSD rats were significantly increased.Acupuncture and sertraline could down regulate their expressions.Sertraline can down regulate the expression of Caspase-3.3.This study suggests that acupuncture treatment of PTSD may be achieved by inhibiting the expression of endoplasmic reticulum stress-related molecules in hippocampus and reducing the expression of apoptosis related factors.This study will provide important scientific basis for clinical acupuncture treatment of PTSD. |
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