| "The spleen corresponding to long summer " means that the spleen in Changxia is in time,and the five internal organs are controlled by the spleen,which together maintain the dynamic balance of the body’s various visceral functional systems.Physiologically,the spleen is compatible with Changxia,transforming everything with soil wet.Pathologically,because the spleen "likes dryness and dislikes dampness","too much moisture" easily hurts the spleen,causing spleen deficiency and dampness.Chronic eczema is a common clinical chronic skin inflammatory disease.Its etiology and pathogenesis cannot be separated from "spleen","dampness" and "changxia".However,the research on its specific mechanism is still blank.This topic is based on The theory of "the spleen should grow in summer" in traditional Chinese medicine and the latest research progress in modern medicine regarding chronic eczema with immune dysfunction and the immune regulation of melatonin.From the perspective of modern medicine regarding melatonin regulating immune function,dry air is the main command As a control in the autumn,we will explore the immunological influence of Moisture Master on the onset of eczema and the intervention effect of Eczema,in order to provide experimental evidence and effective means for the prevention and treatment of seasonal onset of chronic eczema.1 Research purposeBased on the corresponding theory of nature and human of TCM and modern neuroendocrine-immunology,the holistic view of TCM that "spleen should grow in summer" and modern medical immunology are cross-studied.Firstly,we observe the changes in the seasons of summer and autumn on the skin lesions of rats with chronic eczema.Secondly,analyze the correlation between serum melatonin and immune inflammatory cytokines in model rats,reevaluate the intervention effect of Shizhenqing on serum melatonin and inflammatory factors,and finally summarize and explore the relationship between seasonal changes and the onset of chronic eczema Inherent correlation and treatment mechanism of eczema clear.2 Research methods2.1 Theoretical researchFirstly,discuss the theoretical connotation of " The spleen corresponding to long summer",and explore the immune cytokines related to chronic eczema;secondly,analyze the correlation between the material basis of "t The spleen corresponding to long summer " and the immune cytokines related to chronic eczema,again from " The spleen corresponding to long summer " Discuss the influence of seasonal changes on chronic eczema,and combine the study of the formula of Shizhenqing;finally,explore the immunological mechanism of seasonalmelatonin-TH1/TH2-chronic eczema at the theoretical level,and provide the clinical application of Eczema Provide relevant theoretical basis.2.2 Experimental research2.2.1 Season settingThis experiment uses an artificial simulation box to set the average temperature,humidity and light time in the long summer and autumn of Beijing in 2020.The data of temperature and humidity are from China Meteorological Data Network,and the data of day and night light comes from Tiananmen Square in Changxia and autumn.The average time to raise and lower the flag.2.2.2 Experimental animals and model replicationBuy 40 male SD rats of 180-200g,and feed them separately for 5 days in different environments according to 8 rats/cage.The 5 groups of rats are labeled as the control group,the model group and the low,medium,and high concentration eczema groups.The concentration treatment group is hereinafter referred to as the low concentration group,the medium concentration group,and the high concentration group.Except for the control group,the other groups used a pipette to pipette 100 μL of adinitrochlorobenzene-acetone solution with a concentration of 7%on the rat’s back hair removal site for sensitization.A week later,use a 5%concentration of dinitrochlorobenzene-acetone solution.The nitrochlorobenzene-acetone solution was challenged with 200 μL,and then every 5 days,6 times in total.During this period,the photos were taken and recorded and the skin lesions were scored according to the EASI score sheet.After the model was successful,the rats in each group were given intragastric administration.Except that the treatment group was administered at different concentrations,the control group and the model group were given a gavage solution solvent.2.2.3 Index detection①Observe the skin tissues of the control group and the model group under an optical microscope,and take photos for recording.②Record the body weight and spleen weight of the rats weighed on the day of the sampling,and calculate the spleen index.③Separate the serum,determine the concentration of MT,IL-2,IL-4,TNF-γ,and IL-31 in the serum by enzyme-linked immunosorbent method,and determine the concentration of MR in the spleen with BCA protein quantification and enzyme-linked immunosorbent method.The remaining serum is stored in a refrigerator at-80℃.2.2.4 Statistical methodsThe data is analyzed and processed by SPSS 25.0 statistical software.The data results of each group are expressed as mean± standard deviation(x±s).When the control group is compared with the two or five groups of the model group in a single season,if the two groups or the five groups are normally distributed,then use unpaired t-test or one-way analysis of variance,if non-normal distribution or uneven variance,use non-parametric test.3 Research resultsExperiment 1 The influence of the seasonal changes in summer and autumn on the rat model of chronic eczemaSkin EASI score after modeling:(1)Changxia:control group(0 points),model group(2.57 points),low concentration group(2.61 points),medium concentration group(2.38 points),high concentration group(2.59 points),experiment Compared with the control group,the group was statistically significant(P<0.05);(2)Autumn:control group(0 points),model group(2.61 points),low concentration group(2.48 points),medium concentration group(2.59 points),In the high concentration group(2.64 points),the experimental group was statistically significant compared with the control group(P<0.05).The result indicates that the model is copied successfully.The skins were taken uniformly after gavage in the two seasons,and the pathological tissue sections of the skin lesions of the control group and the model group were observed under a 100-fold microscope.The difference:the model group showed a thickening of the thorn cell layer,a large number of inflammatory cell infiltration in the epidermis,and the subcutaneous Typical and obvious inflammatory manifestations of eczema,such as telangiectasia and bleeding,suggest that the two-season chronic eczema model has been successfully replicated at the skin and tissue levels.Experiment 2 The effect of seasonal changes on the serum melatonin(MT),spleen melatonin receptor(MR)and immune cytokines of rats in the control and model groups(1)Seasonal comparison:the serum MT,IL-2,IL-4,IFN-y of the control group showed seasonal rhythms of low in summer and high in autumn(P<0.05);serum MT,IL-2,IL-4 in the model group,IFN-γ showed seasonal rhythms of low in summer and high in autumn(P<0.05).(2)Comparison between groups:IL-4 and IL-31 of the Changxia model group were significantly higher than those of the control group(P<0.05);the autumn model group’s IL-4 was significantly higher than the control group(P<0.05).Experiment 3 Observation of the curative effect of Shizhenqing on chronic eczema model rats in Changxia and AutumnEASI score after Shizhenqing intervention:(1)Changxia:control group(0 points),model group(1.97 points),low concentration group(0.99 points),medium concentration group(1.42 points),high concentration group(1.09 points);and Compared with the model group,the low,medium and high concentration groups all decreased(P<0.05),and the low and high concentration groups were lower than the medium concentration group(P<0.05).(2)Autumn:control group(0 points),model group(1.99 points),low concentration group(0.85 points),medium concentration group(1.29 points),high concentration group(1.04 points);compared with the model group,low,medium,and The high concentration group decreased(P<0.05),and the low concentration group was significantly lower than the middle concentration group(P<0.05).Experiment 4 Exploration of the therapeutic mechanism of Shizhanqing on chronic eczema model rats in Changxia and Autumn(1)Changxia:The serum IL-4 of the model group and the treatment group was higher than that of the control group(P<0.05);the serum IFN-y of the middle concentration group was significantly lower than the control group and the high concentration group(P<0.05);the middle concentration group The serum IL-2 of the model group was significantly lower than that of the control group,the model group and the high concentration group(P<0.05);the serum IL-31 of the model group was significantly higher than the control group(P<0.05),while the eczema clear low concentration group was significantly lower than Model group(P<0.05).(2)In autumn,the serum IL-4 of the model group was significantly higher than that of the control group(P<0.05),while the low,medium and high concentration groups were significantly lower than the model group(P<0.05);serum IFN-y in the low and medium concentration groups It was significantly lower than that of the control group(P<0.05);there was no significant difference in serum IL-2 between the 5 groups(P>0.05);the serum IL-31 of the middle and high concentration groups was significantly higher than that of the control group(P<0.05).4 Research conclusion(1)There are seasonal rhythms of long summer low and autumn high in serum MT and TH1/TH2 cytokines in normal rats and chronic eczema rats(2)The immunological pathogenesis of chronic eczema may be related to the abnormal expression of serum IL-4 and IL-31,and it is more obvious in Changxia(3)The therapeutic effect of Shizhanqing on chronic eczema is not significantly affected by season and concentration,but its immunotherapy mechanism is not the same... |
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