Activation Of Microglia In Anxiety-like Rats With Pathogenic Fire Derived From The Stagnation Of Liver Qi And The Intervention Effect Of Danzhixiaoyao Powder

Objective:Aim to explore the microglial activation in rats with anxiety-liver depression transform fire type and the interventional and the interventional effect of Chinese herbal medicine-Danzhixiaoyao Powder.Methods:First,the chronic restraint stress method was used to establish a rat model of anxiety behavior of liver depression transform fire type from the perspective of etiology.On the 0th day of restraint,the general condition,irritability,tongue color and behavior(elevated plus maze,Open field test)evaluation test of all rats.Screen 36 healthy SD male rats,weighing 230±30g.They were randomly divided into normal group,model group and Danzhixiaoyao Powder group of 12 animals each.Normal group:the same volume of saline as the Danzhixiaoyao Powder group,free diet;Danzhixiaoyao Powder group:continuous restraint for 14 days,,1h/day,the restraint beginning time is random and the Danzhixiaoyao Powder dry cream powder5.0g/kg/d;Model group:continuous restraint for 14 days,1h/day,the restraint beginning time is random,intragastric administration of equal volume of saline.Observe the general condition and weight of the experimental rats during restraint period.After 14 days of restraint,After 14 days of restraint,all animals were subjected to open field experiments and elevated plus maze tests,At the same time,macroscopic indicators such as irritability score and the tongue color were collected to determine whether the model was successfully established.Immunofluorescence was used to observe the polarizing phenotypes M1 and M2 of hippocampus and medial prefrontal cortex iba-1 in each group,as well as CD(16+32)and CD206 in the hippocampus expression;using enzyme-linked immunosorbent assay to detect the relative expression of IL-1β in each group of rats.Results:1.The establishment and evaluation of a rat model of Anxiety-like Rats with Pathogenic Fire Derived from the Stagnation of Liver Qi.2.Before restraint stress:rats in each group are agile,active,chasing and fighting with other rats in the cage,mutual sniffing and mutual assistance,the lips and mucous membranes are well moisturized,the tongue and claws are smooth and pink,and the ears are bright,The corners of the eyes are clean,the secretions are clear,and the body hair is white and bright.There was no violent resistance when grabbing,and normal activity after returning to the squirrel cage.Weight,irritability score,her value of tongue color,behavioral test data and other indicators were balanced(P>0.05).After 14 days of restraint,the model group had messy hair,yellowish dullness,dry lips and mucous membranes,and hard stools.They like to live alone and have a sluggish response,but they are highly nervous and have slow weight gain.Compared with the normal group,the irritability score and the r value of tongue color were significantly increased(P<0.01),the weight gain was significantly reduced,the elevated plus maze experiment tended to exercise in closed arms(OE%,OT% values were significantly reduced),the Open field test activity ability is weakened(the number of crossing grids and the action distance are significantly reduced)(P<0.01).In the Danzhixiaoyao powder group,rats had regular hair,good gloss,red lips and mucous membranes,stools were formed but not dry,mutual help and mutual sniffing in the cage were frequent slightly nervous,but less aggressive than the model group.Compared with the model group,the Danzhixiaoyao powder group had a slow weight gain on days 0-7(P>0.05),but had a rapid weight gain on days7-14(P<0.01),The score of irritability and the r value of tongue were significant decreased(P<0.01 or P<0.05),the elevated plus maze experiment tended to open arm movement(the OE% and OT% values increased significantly)(P<0.01).2.The expression of microglia in each group of rats and the intervention of Danzhixiaoyao Powder(1)The expression of microglia in hippocampus and internal prefrontal cortex After 14 days of restraint,the microglia counts in the hippocampus and medial prefrontal cortex of the model group were significantly higher than those in the normal group(P<0.01);compared with the model group,the cell count of microglia in the Danzhixiaoyao Powder group decreased significantly(P<0.01);(2)Activation of microglia in experimental rats of various groups and intervention effect of Danzhixiaoyao PowderAfter 14 days of restraint,compared with the normal group,the microglia counts in the hippocampus and medial prefrontal cortex of the model group were significantly increased(P<0.01),and the ratio of M1/M2 positive cells in microglia was significantly increased(P<0.01),the expression of IL-1β was significantly increased(P <0.01);compared with the model group,the microglia count in the Danzhixiaoyao Powder group was significantly reduced(P<0.01),and the microglia M1/M2 of positive cells was significantly reduced(P<0.01),and the expression of IL-1β was significantly reduced(P<0.01).Conclusions:1.Anxiety-like Rats with Pathogenic Fire Derived from the Stagnation of Liver Qi have excessive activation of microglia and the shift of phenotype M1/M2 to M1.2.The mechanism of Danzhixiaoyao Powder in the treatment of rats with Anxiety-like Rats with Pathogenic Fire Derived from the Stagnation of Liver Qi may be related to the inhibition of excessive activation of microglia and the correction of the shift of phenotype M1/M2 to M1.