| Depression is a mental illness with low mood as the main clinical feature.It is often clinically manifested as low mood,loss of interest,lack of volition,and negative concepts of decreased digestive function,decreased concentration,self-harm and suicide.Or behavioral symptoms.,So it seriously affects the quality of life of patients.Mongolian medicine believes that excessive stimulation of diet,daily living,and behavior can cause Hey to disorder,cause Hey to become prosperous,and fight with Sheila and Bada,causing the three roots to be out of balance,thereby blocking the conduction of the white veins,and the brain is the white veins.The source,Heyi nourishes the brain through the blood,which leads to depression.As one of the commonly used methods to treat depression,Mongolian medical warm acupuncture(MMWA)therapy has rich literature basis and clinical experience.MMWA therapy uses acupuncture and warm moxibustion stimulation at Dinghui,Heyi,Xin,Stomach,Intestines,etc.,which can effectively regulate blood circulation,dredge white veins,promote digestion and absorb essence,and refresh the mind.Refresh and wait.It is not only safe,no side effects,simple and easy to use,no need to consider the characteristics of drug dependence,etc.,and is widely accepted by patients.Nevertheless,compared with the development of modern medicine,MMWA has only a handful of researches on its mechanism of action,which limits the development and popularization of Mongolian medicine to a certain extent.In recent years,hypotheses such as "neurotrophic factor","hippocampal nerve regeneration",and "brain gut axis" have attracted much attention.This study is based on the above hypothesis of depression,focusing on the core factor of the above hypothesis-brain-derived neurotrophic factor(BDNF)cleavage pathway and the expression of brain-gut peptide SP and VIP.The ProBDNF-tPA-BDNF cleavage pathway is the most important pathway for the cleavage of extracellular BDNF.Therefore,research on the related factors of the lysis pathway and brain-gut peptides will help to better understand the role of BDNF in neuroplasticity,the onset of depression and the resistance to depression.The role of depression in the treatment of depression,at the same time through the intestine-brain axis to achieve the purpose of improving the depressive state and gastrointestinal function of depressed rats.This provides experimental evidence for the clinical use of MMWA to intervene in depression and related diseases.ObjectiveTo explore the role of the changes in the ProBDNF-tPA-BDNF cleavage pathway and brain-gut peptides in the pathophysiological process of depressed rats and the antidepressant treatment of MMWA.The specific mechanism of acupuncture antidepressant treatment provides further experimental evidence.Methods1.Behavioral experiment:90 SPF male SD rats were randomly divided into 6 groups:blank group,CUMS model group,MMWA group,acupuncture group,western medicine group,BDNF blocking group,15 rats in each group.Make a chronic mild unpredictable stress(CUMS)model;comprehensively evaluate the success of model replication through behavioral testing methods such as body weight,sugar water preference,forced swimming,elevated plus maze,and field experiments.2.The above-mentioned behavioral animals used enzyme-linked immunosorbent assay(ELISA)to detect the changes of SP,VIP and other brain-gut peptides in the serum of rats in each group after the treatment.3.Molecular biology experiment:54 SPF male SD rats were randomly divided into 6 groups:blank group,CUMS model group,MMWA group,acupuncture group,western medicine group,BDNF blocking group,9 rats in each group.only.After treatment,detect the expression changes of the important factors ProBDNF,tPA,BDNF and TrkB in the hippocampus by Western blot,and real-time fluorescence quantification was used.The real-time PCR method was used to detect the changes of BDNF mRNA expression in rat hippocampal neurons.4,36 SPF male SD rats were randomly divided into 6 groups:blank group,CUMS model group,MMWA group,acupuncture group,western medicine group,and BDNF blocking group,with 6 rats in each group.Use HE staining The number,morphology and arrangement of hippocampal neurons in rats with depression-like onset and MMWA antidepressant treatment were observed by Honey staining.Result1.Behavioral experimental results:establish a chronic stress depression model,and use open field test,sugar water test,and body weight to comprehensively evaluate whether the model replication is successful.? Weight change:There was no significant difference in the weight of rats in each group before modeling(P>0.05);the weight gain of rats in the CUMS model group was not obvious at all time points after modeling to before taking samples;after treatment,MMWA group The body weight of rats in the acupuncture group and drug group was significantly increased compared with the CUMS model group(all P<0.05),and there was no significant difference between the model group and the BDNF blocking group(P>0.05).? After the 28-day chronic stress program was completed,the intake of sugar and water was significantly lower than that of the normal group.Compared with the model group,the acupuncture group and the MMWA group were higher than the model group,and the model group was blocked by BDNF.There was no significant difference between the groups(P>0.05).The weight change,the number of horizontal crossing grids and the number of erection in the open field test,the acupuncture group and the MMWA group were significantly higher than the model group compared with the model group,the sugar water intake,the acupuncture group,the MMWA group Both were higher than the model group.There was no difference between the model group and the medicine group in the number of horizontal crossing grids and the number of erections in the open field test.The number of horizontal crossing grids and the number of erections in the open field test were lower than those of the normal group.Compared with the model group,the acupuncture group and the MMWA group were significantly higher than the model group.There was no difference between the model group and the drug group.There was no significant difference between the above indicators in the MMWA group(P>0.05).2.Changes in the content of brain-gut peptides:Compared with the blank group,SP and VIP in the serum of CUMS model rats increased and decreased to different degrees with significant differences(all P<0.05).Compared with the blank group,the SP content in the model group,Prozac group,and MMWA group was significantly reduced,with a very significant difference(P<0.01);compared with the model group,the SP content in the MMWA group,acupuncture group,and drug group was obvious There was a very significant difference(P<0.01).Compared with BDNF blockade,there was no significant difference in SP content in the model group(P>0.05).Compared with the blank group,the VIP content in the serum of the model group increased significantly,with significant difference(P<0.05);compared with the model group,the VIP content in the serum of the MMWA group,acupuncture group,and drug group Significantly lower and there is a significant difference(P<0.05),compared with BDNF blockade,there is no statistically significant difference in the VIP content in the serum of the model group(P>0.05).3.MMWA regulates the content of proBDNF-tPA-BDNF related proteins:the expression of hippocampal tPA,BDNF and TrkB protein in the CUMS model group was significantly lower than that of the blank group(P<0.05),and proBDNF in the model group was higher than that of the blank group(P<0.05).Compared with the CUMS model group,the tPA,BDNF protein expression and receptor trkB expression in the MMWA treatment group,acupuncture group,and drug group were significantly increased(P<0.05).and proBDNF was significantly reduced(P<0.05).The expressions of tPA,BDNF and trkB in the hippocampus of rats in the BDNF inhibitor group and the CUMS model group were not significantly different from those in the CUMS model group(P>0.05).In the detection by real-time PCR method,compared with the blank group,the relative expression of BDNF mRNA in hippocampal neurons of CUMS rats was significantly reduced,which was statistically significant.Compared with the model group,the MMWA group The relative expression of BDNF mRNA in the acupuncture group and drug group was significantly increased,with significant differences(P<0.05).4.The influence of Mongolian medicine temperature on neurons:? Analysis of Nissl staining image shows that the pyramidal cells of neurons in CA3 area of the model group are shrunken and incomplete,and the cell arrangement is disordered,the number is obviously lost,and the number of layers is significantly reduced.,Nissl body is lightly stained and partially dissolved.Compared with the blank group,the number of neuron pyramidal cells in the hippocampus CA3 area of the MMWA group,medicine group and acupuncture group was slightly reduced,but compared with the model group,the cell arrangement was orderly and the cell morphology was acceptable.The Nissl body is stained darker.? The analysis of HE staining image shows that the hippocampal cell level of the model group is reduced,the structure is unclear,the pyramidal cell belt and the granular cell belt are disordered,thinned,and interrupted,the neuron membrane is ruptured,the cell atrophys,the nucleus becomes smaller,and there are more nerve cells.There is vacuolization and nuclear pyknosis,and some cells fall off to form vacuoles.The Hippocampus cells of the MMWA group and other groups are arranged more regularly,the cell level is richer,the neuron cell membrane is more complete,but there are still some cells atrophy,nuclei become smaller,and a small number of nerve cells have vacuolization and Nuclear constriction.Conclusion1.The behavioral test results suggest that the improvement of behavioral science proves that the chronic stress depression model is successfully replicated.Chronic stress can cause a decrease in animal exploratory activities,decreased interest,and weight loss.MMWA and acupuncture intervention can improve chronic stress.The depressive behavior of rats has an antidepressant effect.2.MMWA can adjust the content of SP and VIP in the serum of CUMS model rats.Therefore,MMWA can improve the depression of CUMS rats by improving the sensitivity of the intestinal serum brain-gut peptide SP and VIP of CUMS rats.Gastrointestinal dysfunction can improve its depression,and MMWA can improve gastrointestinal function.3.MMWA intervention can significantly regulate the expression changes of ProBDNF,tPA,BDNF and TrkB protein in CUMS model rats.Therefore,MMWA may increase neuroplasticity changes by inhibiting the ProBDNF-tPA-BDNF cleavage pathway,thereby exerting Antidepressant effect.absorb the essence,refresh the brain and achieve a good anti-depressant effect.At the same time,it has the characteristics and advantages of less toxic and side effects,less selection of acupoints,safety and effectiveness. |
PDF Full Text Request