| Objective:At present,there is a lack of research on animal models of influenza wind-heat syndrome.Using wind-heat stimulation and superimposed infection with influenza virus to construct a mouse model of influenza wind-heat syndrome,and to evaluate the model,so as to provide a carrier for the study of related traditional Chinese medicine prescriptions and theoretical mechanisms.Methods:1.Exploration of wind-heat stimulation conditions:36 Kunming mice were randomly divided into normal group,4h group and 8h group according to body weight.From days 1 to 7,the 4h and 8h groups were put into a climate box at 37℃,50% RH,and the second wind speed for wind-heat stimulation every morning.The 4h group was taken out after 4h,and the 8h group was taken out after 8h.The general state of the mice,body temperature,body weight,food intake and water intake were observed every day.The mice were killed on the 8th day.The eyeball blood of mice was taken for routine blood test,and the lung tissue was taken to calculate the lung index.2.Establishment and evaluation of influenza wind-heat syndrome mouse model:72Kunming mice were randomly divided into normal group,wind-heat group,virus group,model group,YQS group and SHJR group,12 mice in each group.From 1 to 7days of the experiment,except for the normal group and the virus group,the other groups were placed in a climate box at 37℃,50% RH,and the second wind speed for8 hours of wind-heat stimulation.On the 8th day,the virus group,model group,YQS group,and SHJR group were anesthetized with isoflurane,and the mouse lung-adapted strain PR8 of influenza virus was instilled into the nose.No wind-heat stimulation was performed after infection,and the other groups were injected with the same amount of normal saline.The general state of the mice was observed and evaluated in the form of symptom scores.Changes in body temperature,body weight,and water intake of mice were monitored.The mice were killed 3 days after viral infection.The blood of mice was taken for routine blood test,the nasal mucosa and lung tissue were taken for pathological observation,and the lungs were weighed to calculate the lung index.Results:1.Exploration of wind-heat stimulation conditions: The excitement and activity of mice in 4h group and 8h group increased.Their tails and paws were redder than the blank control group,and their urine was less frequent and more yellow.The food intake of 4h group and 8h group was less than that of blank control group,and the 8h group was the least.But their water intake was higher than that of the blank control group.The body weight of the 8h group was always lower than that of the blank control group from the 3rd day to the 7th day.The body weight of the 4h group on the 6th and 7th days was lower than that of the blank control group.Lung index,WBC,#NEUT,#MONO,#LYMPH in the 8h group were significantly lower than those in the blank control group and the 4h group.There was no statistical difference between the 4h group and the blank control group.2.Establishment and evaluation of influenza wind-heat syndrome mouse model:(1)General state: On the first day after infection with influenza virus,the model group began to experience rapid breathing,tremors,and messy hair.On the 3rd day after infection,the model group developed rapid breathing,trembling,curling up,and arched back,which lasted for a long time.These mice were less active.The tail and soles of the feet are reddish in color,and some have squinted eyes,and a small amount of secretions appear in the eyes.The performance of the virus group and SHJR group was similar to that of the model group,and the performance of the YQS group was significantly lighter than that of the model group.(2)Characterization integral:On the 1st and 3rd day after infection,the scores of the model group and the virus group were significantly higher than those of the blank control group,and the scores of the YQS group were significantly lower than those of the model group and the SHJR group.There was no statistical difference between the SHJR group and the model group.(3)Weight:On the 3rd day after infection,the body weight of the model group and virus group was lower than that of the blank control group,the body weight of the YQS group was significantly higher than that of the model group,and there was no statistical difference between the SHJR group and the model group.(4)Water intake,food intake:After wind-heat stimulation and on the first day after infection,the water intake of each stimulation group was higher than that of the blank control group.On the 2nd day after infection,the water intake of the model group was lower than that of the blank control group,but higher than that of the virus group;On the 1st and 2nd day after infection,the food intake of the virus group and the model group was lower than that of the blank control group,and the YQS group was higher than the SHJR group and the model group.(5)Body temperature: On the 3rd day after infection,the temperature in the model group and virus group was significantly lower than that in the blank control group,and the temperature in the YQS group was significantly higher than that in the model group and SHJR group.There was no statistical difference between SHJR group and model group.(6)Blood routine: WBC,#NEUT,#MONO in virus group and model group were significantly higher than those in blank control group;WBC and #NEUT in YQS group were significantly lower than those in model group;There was no statistical difference between SHJR group and model group.(7)Lung indext,lung index inhibition rate: The lung index of the virus group and model group was significantly higher than that of the blank control group,and the YQS group was significantly lower than the model group and SHJR group.There was no statistical difference between SHJR group and model group.The lung index inhibition rate in the YQS group was higher than that in the SHJR group,and the lung index inhibition rate in the SHJR group was negative.(8)Pathology of nasal mucosa: The blank control group had no obvious lesions;mucosal epithelial cells in the wind-heat group were disordered,with a small amount of cilia shedding,and no inflammatory cell infiltration;in the virus group,there was a large amount of inflammatory cell infiltration in the lamina propria of the nasal mucosa;Epithelial cells in the model group were partially shed,disorderly arranged,and had obvious inflammatory cell infiltration;the epithelial cells in the YQS group were neatly arranged,the cilia were not shed,and the inflammatory cells were significantly less;The SHJR group was the most severe,with more epithelial cell shedding and extensive inflammatory cell infiltration.(9)Pathology of lung tissue: There were no obvious lesions in the lung tissue of the blank control group and the wind-heat group;the alveolar wall of the virus group was widened,and there was obvious inflammatory cell infiltration;The alveolar structure of the model group was incomplete,the alveolar wall was widened and thickened,and there was a large number of inflammatory cell infiltration;the alveolar structure of the YQS group was clear and complete,and no obvious inflammatory cell infiltration was found;the SHJR group was similar to the model group.Conclusion:The mouse model established by wind-heat stimulation(37℃,50%RH,sec ond-speed wind speed,8h stimulation per day,7 days in a row)+ PR8 influen za virus intranasal instillation has obvious flu symptoms and pathological manif estations,except for no fever,the rest of the manifestations have the characte ristics of wind-heat syndrome.Yinqiaosan treatment can significantly improve th e model performance.Sanhanjiere oral liquid treatment did not improve the mo del,but aggravated the damage of the nasal mucosa and lung tissue of the mo del.Based on the results of the test and verification,it was proved that the syn drome type of the model belonged to the wind-heat syndrome.the model was s uccessfully constructed. |
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