| Research background: The gastrointestinal tract is the body’s digestive organ and the largest immune organ.The gastrointestinal tract not only has the functions of digestion and absorption of food,but also has the functions of hormone secretion,immune regulation and mucosal barrier.Existing studies have found that there is an obvious correlation between human intestinal bacteria and the immune system of the gastrointestinal tract.The former can trigger the immune inflammatory reaction of the human intestinal tract,especially the genetically susceptible people will have intestinal or extragastrointestinal allergic diseases,such as atopic dermatitis.Atopic dermatitis(AD),also known as heterotopic eczema and genetic allergic eczema,is a recurrence inflammatory skin disease related to heredity and immunity.Epidemiological studies have shown that the incidence of AD is positively correlated with the development of national economy and industrialization,especially the change of dietary structure.Now scholars have confirmed that oral probiotics can affect the intestinal immune regulation mechanism of AD,and can relieve the allergic symptoms such as pruritus.TCM believes that the onset of AD is caused by dampness and heat of dampness and stagnation of spleen deficiency.The acute stage is the spleen loss of health,the spleen loss of department of transportation and transformation,and the dampness and turbidity accumulation.The chronic stage is the spleen deficiency,blood dryness and skin loss of nourishment.Therefore,it is considered that the weakness of spleen and stomach is the physical characteristic of AD patients,dampness is an important pathological factor of the onset of atopic dermatitis,and the restoration of spleen function is the key to the treatment of AD.The prescription of invigorating spleen,nourishing blood and dispelling wind is an empirical prescription for the treatment of atropic dermatitis in our hospital.The prescription invigorates spleen by nourishing qi and blood,moistens skin cou,dispelling wind and relieving itching and improves skin barrier function.Purpose and Significance: Traditional Chinese medicine decoction is mostly absorbed through the intestinal barrier,so we thought that the prescription of strengthening spleen,nourishing blood and removing wind may improve intestinal immune function and activate immune signals by regulating intestinal flora of AD patients,thus causing multiple intestinal links to synergistic regulate the immune and anti-inflammatory effect of Treg cells in CD4+T cells.In this study,the possibility of activating spleen,nourishing blood and dispelling wind by regulating intestinal fragility Bacteroides to regulate immune cytokines to improve the mechanism of AD was explored.To further provide a scientific clinical basis for the treatment of spleen deficiency atopic dermatitis with the prescription of invigorating spleen,nourishing blood and quefeng.It provides a new idea for the clinical application of the prescription of invigorating spleen,nourishing blood and dispelling wind and the clinical treatment direction of spleen deficiency atopic dermatitis.Methods:(1)Patients with spleen deficiency atopic dermatitis in outpatient and inpatient departments of our hospital were collected,and certain healthy volunteers were recruited.Patients were randomly divided into the spleen nourishing blood functions of the group of bifidobacteria and butyric acid bacteria duplex living bacterium group,positive control group),observe the oral spleen nourishing blood functions of party by bacteroides fragilis skin symptoms and immune function,improve AD reveals the regulation by intervening in the intestinal flora,possible mechanisms of alzheimer disease AD process;(2)A mouse model of AD with intestinal flora disorder was established by immune damage and antibiotic gastric administration to study the mechanism of the intervention of spleen nourishing blood quefeng prescription in the development of AD by regulating Bacteroidetes fragilis,and to provide a theoretical basis for the treatment of AD by spleen nourishing blood quefeng prescription through intestinal immune function.Results:1.Experiment 1:(1)Comparison of symptoms in the traditional Chinese medicine group after treatment showed that P values in each week were all less than 0.05,which was statistically significant.It can be seen that all symptoms of the patients were improved after oral treatment with the prescription of invigorating spleen,nourishing blood and dispelling wind,and the symptoms were significantly improved after 4 weeks of treatment.(2)Comparison of symptoms in the probiotics group after treatment showed that P values in each week were all less than 0.05,which was statistically significant.It can be seen that after oral probiotics treatment,all the symptoms of the patients have improved,and the symptoms improved significantly after 8 weeks of treatment.(3)The comparison of SCORAD scores between the TCM group and the probiotics group before and after treatment showed that before treatment,t=-1.316,P=0.263>0.05,indicating that there was no significant difference between the two groups before treatment and there was no statistical significance.4 weeks after treatment,t=7.375,P=0.000<0.01,indicating that the symptoms of the two groups of patients improved significantly after 4 and 8 weeks of treatment,and the TCM group was better than the probiotics group in improving SCORAD scores.(4)Fecal SIg A expression in the TCM group and the probiotics group before and after treatment: using rank sum test,Z=-1.174,P=0.240,the difference was not statistically significant(P>0.05).After treatment,independent sample t test showed that t=-34.560,P=0.000,and the difference was statistically significant(P<0.05).There was significant difference in the level of SIg A between the two groups after treatment,and the TCM group was significantly better than the probiotics group in improving the level of high SIg A.(5)The expression of serum Ig E level in the TCM group and the probiotics group before and after treatment: independent sample t test was used.Before treatment,t=-0.528,P=0.599,and the difference was not statistically significant(P>0.05).After treatment,t=4.987,P=0.000,the difference was statistically significant(P<0.05).Combined with Table 5,it can be seen that the two groups had significant difference in serum total Ig E after treatment,and the TCM group was significantly better than the probiotics group in reducing serum total Ig E.(6)TGF-β expression levels in the TCM group and the probiotics group before and after treatment: independent sample t test was used.Before treatment,t=-0.592,P=0.556,the difference was not statistically significant(P>0.05).After treatment,t=-11.256,P=0.000,the difference was statistically significant(P<0.05).The TGF-βlevel was significantly different between the two groups after treatment,and the TCM group was significantly better than the probiotics group in promoting the high TGF-βlevel.(7)Fluorescence quantitative PCR of Bacteroides vulnerability in feces:Rank-sum test was used.Before treatment,Z=-0.165,P=0.869,and the difference was not statistically significant(P>0.05).After treatment,Z=-6.698,P=0.000,the difference was statistically significant(P<0.05).After treatment,the abundance of Bacteroidetes fragilis was significantly different between the two groups.The abundance of Bacteroidetes fragilis in the Chinese medicine group was significantly better than that in the probiotics group.2.Experiment 2:(1)After administration,body weight of mice in each group increased compared with before the experiment,with statistical significance(P<0.05);Compared with blank group,the body weight of mice in experimental groups was significantly lower,and the difference was statistically significant(P<0.05).(2)Score of back skin damage of mice The healing rates of mice in the high-dose,medium-dose and low-dose treatment groups of invigorating spleen,nourishing blood and qufeng prescription and the oral probiotics control group were about 100%,85.71%,71.43% and 71.43%,respectively.Compared with the model group(group A),the other groups had significant curative effects,with statistical significance(P<0.000).(3)Compared with model group(group A),the serum Ig E level of mice in positive control group(group P),high-dose and low-dose groups of invigorating spleen,nourishing blood and dispelling wind was significantly decreased,with statistical significance(P<0.001).(4)Compared with model group(group A),the serum IL-17 A level of mice in the high-dose group of invigorating spleen and nourishing blood and qufeng prescription and the positive control group was significantly decreased,the difference being statistically significant(P<0.01);the serum IL-17 A level of mice in the medium-dose and low-dose groups of invigorating spleen and nourishing blood and qufeng prescription was significantly decreased,the difference being statistically significant(P<0.05).(5)Compared with group A,except the blank group(NC group),the level of fecal s Ig A in other groups was significantly increased,with statistical significance(P<0.001).(6)Compared with group A,the serum TGF-1 level of mice in the high-dose group of invigorating spleen,nourishing blood and qufeng prescription and the positive control group(group P)was significantly increased(P<0.001),and the serum TGF-1 level of mice in the medium-dose group of invigorating spleen and nourishing blood and qufeng prescription was significantly increased(P<0.05);There was no significant change in serum TGF-1 level of mice in the low-dose group of invigorating spleen,nourishing blood and qufeng prescription(P>0.05).Compared with the blank group(NC group),the serum TGF-1 level of mice in model group(A group)was significantly decreased,with statistical significance(P<0.01).(7)Compared with NC group,the expressions of MUC2(rich in mucin-2),Occludin(tight junction protein)and ZO-1(closed protein)in group A were significantly decreased(P<0.001).Compared with model group A,the m RNA expression levels of MUC2,Occludin and ZO-1 in the low-dose and medium-dose groups of invigorating spleen,nourishing blood and qufeng prescription were all increased,but there was no significant difference.Compared with group A,the expression levels of MUC2,Occludin and ZO-1 in the high-dose group of invigorating spleen,nourishing blood and dispelling wind prescription and positive P group were all increased,and the difference was statistically significant(P<0.05).(8)Compared with the blank group(NC group),the abundance of Bacteroides Fragilis in group A was significantly decreased(P<0.001).Compared with the model group(group A),the relative abundance of Bacteroides Fragilis in the probiotics group and the high-dose Spleen-nourishing Blood Qufeng Formula group increased significantly,with statistical significance(P<0.01).After treatment,the relative abundance of Bacteroides Fragilis in each group was still lower than that in the blank group.Conclusion:1.Experiment 1:In the treatment of spleen deficiency atopic dermatitis,both the traditional Chinese medicine group and the probiotics group could significantly relieve the skin lesions of the patients,showing obvious clinical efficacy.During the 8-week treatment,it was found that the intervention effect of the traditional Chinese medicine group in the early treatment was obvious.The probiotics group also significantly improved the symptoms of patients with spleen deficiency atopic dermatitis in the long term.By observing its blood immunological indexes,we can know that the TCM group is obviously better than the probiotics group in treatment.In the observation of intestinal immunity,it was found that the prescription of invigorating spleen,nourishing blood and dispelling wind had the same effect as probiotics in improving intestinal immune barrier.It was also found that the relative abundance of Bacteroidetes fragile in the intestinal tract of AD patients was lower than that of healthy people,but it was significantly increased after treatment.It can be concluded that the prescription of invigorating spleen,nourishing blood and dispelling wind can regulate the relative abundance of Bacteroides fragilis in the intestine to improve the function of intestinal immune barrier.2.Experiment 2:(1)After administration,the body weight of mice in each group increased compared with that before the experiment,indicating that the traditional Chinese medicine invigorating spleen,nourishing blood and dispelling wind prescription and probiotics can both improve the spleen and stomach function of mice,relieve diarrhea,loose stools and other symptoms of spleen and stomach weakness,and increase the body weight of mice.(2)The epidermal skin lesions of mice in P group(positive control group)were significantly alleviated,while those in the high-dose group of invigorating spleen,nourishing blood and qufeng formula had significant effect on the improvement of skin lesions.The pathological changes of the epidermal lesions in mice were positively correlated with the back skin score.Both the Chinese medicine group and the probiotics control group could improve the epidermal inflammatory response of AD mice with spleen deficiency and reduce itching and other symptoms,and the overall efficacy of the Chinese medicine group was better than that of the probiotics group.(3)Both invigorating spleen,nourishing blood and quelling wind prescription and probiotics can regulate the balance of intestinal flora,improve intestinal spleen and stomach functions,effectively reduce the high expression of serum Ig E in the body,inhibit allergic reaction,and have a significant effect on mice with spleen deficiency atopic dermatitis.(4)From the observation of the expression levels of serum IL-17 A and TGF-1cytokines in mice,it was found that the high dose group of TCM showed obvious advantages in exerting anti-inflammatory and immunomagulatory effects.Considering the presence of centipede in traditional Chinese medicine,centipede has certain toxicity.The subsequent analysis of the effect of toxicity component index of Chinese medicine on therapeutic index should be added.(5)By observing the protein m RNA expression levels of SIg A,which reflects intestinal immune function,and intestinal mucosal barrier,in conclusion,the prescription of invigorating spleen,nourishing blood and dispelling wind and the probiotics group can repair intestinal barrier damage and intestinal homeostasis imbalance in AD mice,rebuild intestinal immune balance,and promote intestinal function recovery.(6)The prescription of invigorating spleen,nourishing blood and dispelling wind can regulate the relative abundance of Bacteroides fragilis in the intestines. |
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