| Purpose:In this experiment,under the guidance of the principle of treatment of diseases in traditional Chinese medicine,the rat model of liver depression and spleen deficiency depression was induced by chronic stress stimulation,moxibustion at Baihui,Taichong,Ganshu and Zusanli points,and rat body weight and behavioral changes were assessed.Models,and determination of rat hippocampal tissue morphology and CaMKII protein content and gene expression,to explore the effect of moxibustion prevention and treatment on depressed model rats with liver depression and spleen deficiency,to provide experimental basis for clarifying the mechanism of moxibustion in preventing and treating depression,Provide new ideas for the clinical application of moxibustion therapy to prevent and treat depression.Material and method:72 SPF rats were randomly divided into six groups:normal group,model group,moxibustion prevention group,fluoxetine prevention group,moxibustion treatment group,and fluoxetine treatment group,12 in each group.A rat model of liver depression and spleen deficiency was prepared by using chronic stress stimuli.Rats were randomized to one of the following six types of stimuli on a daily basis,and chronic unpredictable stress stimuli were taken.Six groups of stimuli were:(1)fasting,water banned 12h;(2)day and night reversed;(3)45℃ high temperature hot water swimming dedicated to observe the swimming status of each rat;(4)long tail clip folder tail 5min;(5)11℃ cold water swimming to exhaustion observation of rat state;(6)45℃ Tilt feeding for 12 hours.once daily stimulation,the same stimulating factor does not appear continuously,so that the mice can not predict the occurrence of stimulation.Each stimulus was randomly arranged,accumulated 3 times,and a total of 18 days of modeling time.Using weight changes and field tests to confirm the success of the model.After successful modeling,the rats were treated with moxibustion and western medicine.The moxibustion intervention method was selected:Baihui,Ganshu,Zusanli and Taichong points.Moxa moxibustion with a diameter of 0.7cm and a length of 3.5cm was applied to each hole.Moxibustion time is 15 min.Fluoxetine gavage method:According to the body surface area exchange algorithm,rats were given a concentration of 0.2mg/mL,intragastric volume of 1ml/100 g per day.Normal group:free drinking water,no stimulation,normal feeding.Model group:daily chronic stress modeling.In the moxibustion prevention group and fluoxetine prevention group,in addition to the daily chronic stress stimulation,the moxibustion prevention group was given prevention and treatment from the 7th to the 18 th day before the modeling;the fluoxetine prevention group was from the 7th-18 d was given intragastrically 1 hour before modeling.Moxibustion treatment group:Moxibustion was given from the 19 th to the 30 th day after the model was completed.Fluoxetine treatment group:The rats were given intragastric administration from the 19 th to the 30 th day after the completion of the model.The morphology of rat hippocampus was observed by electron microscopy.The expression of CaMKII and p-CaMKII protein in hippocampus was detected by Western blot.The expression of CaMKII gene in hippocampus was detected by RT-PCR.Results: 1.The behavioral changes in rats:Before the experiment,there was no significant difference in the behavior of rats in each group.The daily growth rate of rats after the end of the experiment,the number of horizontal crossings in the open field experiment and the normal number of vertical climbs were higher than those in the model group,with significant differences(P<0.05);indicates successful modeling.Moxibustion prevention group,moxibustion treatment group,fluoxetine prevention group,fluoxetine treatment group rats were all higher than the model group in the above aspects,with significant differences(P<0.05).2.Electron microscopic observation of hippocampal morphology in rats:In the normal group,the astrocytes in the hippocampus were spindle-shaped,nucleus was round and light,and the cytoplasm was full.The rough endoplasmic reticulum and mitochondria were seen.In the model group,the astrocytes were spindle-shaped,with round and light nucleus and light cytoplasm.The apical endoplasmic reticulum and mitochondria with loosening of mitochondria were obviously expanded.In the moxibustion prevention group,the astrocytes were spindle-shaped,and the nucleus was elliptical,light,and full of cytoplasm.The rough endoplasmic reticulum,mitochondria,and ribosomes were seen.In the fluoxetine prevention group,the astrocytes were spindle-shaped,the cytoplasm was relatively vague,and the nucleus was elliptical and light,showing mitochondria.In the moxibustion treatment group,the astrocytes were spindle-shaped,the nucleus was elliptical,light,and the cytoplasm was full.The rough endoplasmic reticulum,mitochondria,and ribosome were seen.In the fluoxetine-treated group,the astrocytes were fusiform,the cytoplasm was relatively vague,and the nucleus was elliptical and light.Visible mitochondria,ribosomes,and other organelles,indicating that it was damaged.3.Compared with normal group,the expression of CaMKⅡprotein in hippocampus of rats:Compared with the normal group,the expression of CaMKII mRNA in the depressive model group was significantly lower(P<0.05);the other five groups were significantly different(P<0.05),the moxibustion prevention group was the most significant;compared with the depression model group,moxibustion prevention group,fuoxetine prevention group,moxibustion treatment group,fluoxetine treatment group rats mRNA expression were significantly increased(P<0.05);compared with moxibustion prevention group,fluoxetine prevention group,in the moxibustion treatment group,the mRNA expression of the rats in the fluoxetine-treated group was significantly lower(P<0.05),and the expression of mRNA in the fluoxetine-treated group was significantly lower than that in the moxibustion-treated group(P<0.05).4.Expression of morphological CaMKII protein in rat hippocampus:Compared with the normal group,the expression of CaMKII protein in the depression model group was significantly lower(P<0.05),and the ratio of p-CaMKII/CaMKII was significantly lower(P<0.05);compared with the depression model group,moxibustion prevention group,fluorine The expression of CaMKII protein was significantly increased(P<0.05)and the ratio of p-CaMKII/CaMKII was significantly increased(P<0.05)in the Xiting prevention group,the moxibustion treatment group,and the fluoxetine treatment group.Compared with the fluoxetine-treated group,the moxibustion-treated group,and the fluoxetine-treated group,the protein expression and the phosphorylation ratio were significantly decreased(P<0.05);compared with the moxibustion-treated group,the fluoxetine treatment group was larger.The expression of rat protein and its phosphorylation ratio were relatively high(P<0.05).Conclusion: 1.Moxibustion produces the same antidepressant effect as fluoxetine.2.Moxibustion can specifically increase the expression of CaMKⅡ gene in hippocampus of rats with liver-qi deficiency and spleen-qi deficiency,promote the phosphorylation of CaMKⅡ protein,play a significant role in the treatment and delay the occurrence and development of depression. |
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