Experimental Study On The Effect Of Acupuncture At Quchi, Xuehai And Zusanli Points On PI3K/AKT Signaling Pathway In Urticaria Mice

Purpose:To observe the effects of acupuncture at Quchi(L11),Xuehai(SP 10),Zusanli(ST36)points in mice models with urticaria(Type I allergy model)during the degranulation of mast cells,by analyzing the changing expressions of phosphatidylinositol-3-kinase(PI3K),protein kinase B(AKT)signaling pathways and compare the changes of serum immunoglobulin E,Ig E,investigates the mechanism and effectiveness of acupuncture on the regulation of urticaria by morphology and molecular biology.Material and method:40 male ICR mice(8weeks)were choose for the test.The weight of mice was 20-25 g.Randomly divided into 4 groups(10 mice for each group): normal control group,model control group,acupuncture group(L11,SP10,ST36)and positive drug group.In the experiment,PCA was performed for the mice model with urticaria.Acupuncture group was acupunctured at L11,SP10,ST36 point before the experiment start.The positive drug group was given 0.2mg of loratadine by gavage.Acupuncture group and the positive drug group were treated 7days for each group.The model of passive cutaneous anaphylaxis was established on the 8th day.After mice were anesthetized with ether,except for normal control group,0.1ml phosphate solution containing 0.5% anti DNP Ig E was injected into the right ear skin of mice in the other three groups.In the normal group,0.1ml 0.9% sodium chloride solution was injected into the skin at the same position.After 48 hours,four groups were attacked with antigens.The tail vein was injected with 0.9% phosphate solution containing DNP-HSA 0.5 ? g/ ? l and 1% Evans blue(200 ? l).After the successful establishment of the model,the aorta abdominalis blood and sensitized ear skin tissue of mice was taken for the test.(1)By using HE staining,detect the histomorphology of mice skin in the sensitized area(2)Using ELISA to observe the effect of serum Ig E level in mice(3)The effects of PI3 K / P-PI3 K,AKT/P-AKT protein expressions in mice were observed by immunohistochemistry and analyzed statistically.Results:1.HE staining of mice ear skinUnder the microscope,the structure and the shape of epidermis and dermis of normal control group were intact,the thickness was normal,the cells arranged orderly,the structure was clear,and no edema was found.In the model control group,there were severe edema and inflammation in the epidermis and dermis under the microscope.In the epidermis,there were severe keratinization,and granular layer being thickening,cells were destructed in the basal layer.Various inflammatory cells were infiltrated in the dermis,and the boundary between the cells was not clear.In the positive drug group,the epidermis and dermis were slightly thickened,the cuticle was slightly keratinized,the boundary between cells was clear than model control group,the structure was basically complete,showing mild edema and inflammatory reactions.The acupuncture group has no obvious damage to the structure.It improves the inflammatory infiltration of the skin tissue in the sensitized area and restores the structure and shape of the tissue.2.The observation of serum Ig E level in mice from each group The level of total Ig E in serum of model control group was significantly higher than normal control group(P < 0.05).The results showed that the model of urticaria in mice was successful.The level of total Ig E in mice serum of acupuncture group and positive drug group was significantly lower than model control group(P < 0.05).The results showed that both groups could control the level of serum Ig E in mice with urticaria.The serum total Ig E level of the positive drugs group was lower than the acupuncture group(P < 0.05).3.The observation of the expression from PI3 K,P-PI3 K,AKT,P-AKT protein in mice sensitized skin tissue The protein expression level of PI3 K,P-PI3 K,AKT,P-AKT in model control group was higher than normal control group(P < 0.05).The protein expression level of PI3 K,P-PI3 K,AKT,P-AKT in acupuncture group and positive drug group was lower than that in model control group.There was no statistical significance in PI3 K between the two groups and model control group,but P-PI3 K,AKT,P-AKT had statistical significance(P < 0.05).The protein expression level of PI3 K,P-PI3 K,AKT,P-AKT in the positive drugs group was lower than that in the acupuncture group,P-PI3 K,AKT,P-AKT had statistical significance(P <0.05).Conclusion:1.Acupuncture at Quchi(L11),Xuehai(SP10)and Zusanli(ST36)points can reduce the level of total serum Ig E.It proves that acupuncture can control the Type I allergy hypersensitivity.2.Acupuncture at Quchi(L11),Xuehai(SP10)and Zusanli(ST36)points can regulate PI3 K and AKT signal pathways,it inhibits the degranulation of mast cells and release of inflammatory mediators by inhibiting the phosphorylation of PI3 K and AKT,which proves that acupuncture can prevent urticaria.