| Objective:To explore the effect of alternate stimulation of cold and heat on the immune function of lung and skin of rats and the intervention effect of Yupingfeng Powder on the function of protecting body from the outside of the body,and to provide experimental basis for improving the immune defense function of the body.Methods:1?Subgroups:108 healthy male Wistar rats of SPF grade,weighing 180-220g,were divided into 6 groups according to random number table,18 rats in each group.The results were as follows:group A in normal group,group B in alternate group of cold and heat,group C in cold and heat alternately with Chinese medicine intervention group,group D in group D of low immunity from normal temperature,group E in alternating group of cold and heat with low immunity,and group F with alternating cold and heat of low immunity plus intervention group of traditional Chinese medicine.2?Modeling and administration:1)Establishment of immunocompromised rat model:In addition to the normal group,the immunocompromisedratswereinjectedsubcutaneouslywithhydrocortisone20mg/(kg·d)for 5 days.2)the model of rats stimulated by cold and heat alternately:First,the rats were given cold stimulation,and the artificial climate box RH(%)(50±4),(0±2)?,40 minutes per day was set up.After the cold stimulation,the rats were given thermal stimulation,and the artificial climate boxRH(%)(50±4),(33±2)?was set up,which was stimulated for 20minutes per day.3)Drug intervention:After the rats were exposed to cold and heat alternately in an artificial climate box every day,the daily dose of Yupingfeng Powder was converted into 25g/kg of rats according to the equivalent conversion method of the dosage of traditional Chinese medicine.At the same time,rats in other groups were given distilled water at the corresponding dose.3?Testing:(1)Generalobservation:observeandrecordthementalstate,activity,coat color,joint,stool,intake,weight and so on.(2)morphological observation:The pathomorphological changes of lung and skin tissues of rats in different periods were observed by HE staining and light microscope at the 7th day and 14th day and 28th day respectively.(3)Immunoassay:The IgG content of immunoglobulin in tracheal lung lavage fluid(airway fluid)was measured at 7d,14d,28d,respectively.The levels of CD4~+,CD8~+and IL-1?and TNF-?in the lung and skin of rats were measured.(4)The wet/dry weight ratio of left lung was measured.Results:1?General conditions:Compared with the normal group,the mental,activity,coat color,joint,stool,water intake and body weight of the rats in each model group changed in different degrees on day 7,day 14,and day 28,respectively.Show a series of pathological symptoms.During the course of the experiment,the rats showed signs of mental depression,decreased activity,curling up,piling up,increasing nasal secretions,disorderly back hair,dilute stools,reduced intake and drinking water,and slow weight gain after the rats were subjected to alternate cold and heat stimulation.Some of the rats showed foot dorsal edema with erosion and ulceration,joint swelling and other pathological signs.With the prolongation of intervention time,animal behavior disease Rational performance is gradually increasing.2?Morphological observation:The pathological changes of lung and skin of rats were observed by HE staining.Compared with the normal group,the cold-heat alternation group and the low-temperature-free cold-heat alternation group had different degrees of damage tochronicinflammatorycellinfiltration,lymphocyteinfiltration,fibroustissue proliferation,alveolar atrophy and so on.However,the degree of lesion of skin tissue was not obvious,or only a small number of lymphocytes infiltrated in the skin tissue of the low-model group,and the results showed that the alternating stimulation of the cold and heat was more obvious to the lung tissue.3?Immune index T cell subgroup CD4~+and CD8~+:Compared with normal group,cold and heat alternate group,low immune normal temperature group,low immune normal temperature group,The contents of CD4~+and CD8~+in lung and skin of rats with low immunity were significantly decreased(P<0.01),and the decrease of indexes in lung and skin tissue was most significant in low immunity model group,compared with that in cold and heat alternate group.The contents of CD4~+and CD8~+in lung and skin of rats in low immunity cold and heat alternation group were lower than those in low immunity cold and heat alternating group(P<0.01),and the contents of CD4~+and CD8~+in lung and skin of rats in cold and heat alternate group were higher than those in low immunity cold and heat alternate group,and in low immunity cold and heat alternate group,low immunity cold and heat alternate group was added in medium and low immunity.The contents of CD4~+and CD8~+in lung and skin of rats in the drug intervention group were increased(P<0.01).The changes of CD4~+CD8~+in lung and skin tissues of rats were consistent at 14 days and 28days,respectively.4?Compared with the normal group,the contents of IL-1??TNF-?in lung and skin of rats in the cold and heat alternate group,low immunity normal temperature group and low immunity cold heat alternating group were lower than those in the normal group.The decrease of index content in lung and skin tissue of low immune model group was the most significant(P<0.01).The contents ofIL-1??TNF-?in lung and skin of rats in low immunity cold and heat alternant group were all decreased(P<0.01),and the contents of IL-1??TNF-?in lung and skin of rats treated with Chinese herbal medicine were significantly increased(P<0.05 or P<0.01).The contents of IL-1??TNF-?in lung and skin of rats in the control group were significantly higher than those in the control group(P<0.01),and the contents of TNF-?in the lung and skin of the rats in the control group were significantly higher than those in the control group(P<0.01).The contents of TNF-?in the lung and skin of rats in the control group were significantly higher than those in the control group.The change trend of IL-1??TNF-?in lung and skin tissues of rats was consistent.5?Immunoglobulin IgG in trachea and lung lavage fluid(short for airway fluid):Compared with normal group,the content of IgG in airway fluid of rats in cold and heat alternate group,low immunity normal temperature group and low immunity cold heat alternate group were all significantly decreased,and the results were statistically analyzed.There was significant difference(P<0.01),compared with the low-immunity normal temperature group,the content of IgG in the airway fluid of the rats in the low-immunity group was significantly lower than that in the low-immunity group(P<0.01),and the content of IgG in the airway fluid of the rats in the low-immunity group was significantly lower than that in thegroupofcoldandheatalternatinggroup(P<0.01).Thedifferencewas significant(P<0.01).For the intervention combined with traditional Chinese medicine group increased airway fluid IgG in varying degrees,with statistical significance(P<0.05 or P<0.01);and low free and alternating group compared to low free and alternating with traditional Chinese medicine intervention group rats airway fluid IgG content was significantly increased,with significant difference(P<0.01).6?On the 7th day,compared with the normal group,the cold and heat alternate group,the low immunity normal temperature group and the low immunity cold heat alternating group,the left lung wet dry weight ratio(W/D)of the rats in the left lung wet dry weight ratio was significantly higher than that in the normal group(P<0.01);on the 14th day,there was a significant difference between the two groups(P<0.01).Compared with the normal group,the wet and dry weight of left lung in the low immunity group was higher than that in the control group(P<0.05).Conclusion:1?Hydrocortisone could decrease the immune function of rats.2?The general condition(mental activity,skin color,joint,nasal secretions,feces,intake and drinking water,body weight,immune index(CD4~+,CD8~+,IL-1?,TNF-?content in lung and skin)of rats stimulated by alternate cold and heat stimulation,The content of immunoglobulin(IgG)in airway fluid,the wet/dry weight ratio of left lung(W/D)and the histomorphology were all significantly affected,and the effects of alternately cold and heat stimulation were more obvious in the hypoimmune rats than in the normal group.3?GeneralinformationofYupingfengPowderonrats(mentalactivity,skin color,joint,nasalsecretions,feces,intakeanddrinkingwater,bodyweight,immune index(CD4~+,CD8~+,IL-1?,TNF-?content in lung and skin,The content of immunoglobulin IgG in airway fluid and the ratio of wet to dry weight of left lung(W/D)and histomorphology were significantly improved and immunomodulatory effect of Yupingfeng Powder was better than that of low immune group.4?On day 7 and day 14 and at different periods of 28 days,the changes of immunological index CD4~+,CD8~+,IL-1?and TNF-?in lung tissue and skin tissue of rats were consistent,and it was found that under the condition of alternate cold and heat stimulation,the content of TNF-?in lung tissue was similar to that in skin tissue.There is a strong correlation between the lung and the skin in the pathological changes,which is consistent with the classical explanation of"extrapulmonary fur". |
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