Anti-allergic Effect Of Traditional Dai Prescription, Ya-Jie-Sha-Ba,and Traditional Chinese Medicine, Sophorae Flavescentis Radix

OBJECTIVE: Traditional Dai prescription Ya-Jie-Sha-Ba is wildly used in Traditional Dai Medicine for the treatment of food allergy and food intolerance.However, information about Ya-Jie-Sha-Ba’s active ingredients, effects, and mechanisms of action are limited. This research aimed to establish a method for determination of palmatine and berberine in Ya-Jie-Sha-Ba and elucidate the effects and underlying mechanism of Ya-Jie-Sha-Ba prescription in the treatment of food allergies through a Balb/C female mice sample with food allergy. On the other hand,immunoregulatory effects of Sophorae Flavescentis Radix(SFR), were explored on allergic dermatitis mice induced by dinitrofluorobenzene(DNFB).The METHODS:1.To find the extraction method of palmatine and berberine in Ya-Jie-Sha-Ba by orthogonal text and to establish a method for determination of palmatine and berberine in Ya-Jie-Sha-Ba.2.A food allergy model was established in female BALB/c mice by injecting ovalbumin(OVA) three times at 0 h, 48 h, and 96 h. OVA-sensitized mice were supplemented with no treatments(control), Ya-Jie-Sha-Ba, loratadine, palmatine, or berberine. Scratching frequency, serum Ig G, Ig E, IL-4, IL-10, IL-17, IL-21, IFN-γ,and TNF-α amounts were measured at 50 h and 98 h. Regulatory T cells(Tregs)percentages were evaluated by flow cytometry at 98 h.3.Allergic dermatitis mice were established on male Kunming mice induced by DNFB painting. DNFB-sensitized mice were administered with SFR(low and high dosage), oxymatrine(low and high dosage) and loratadine, respectively. Scratching frequency, ear thickness, the indexes of thymus and spleen, serum IL-2、IL-6、IL-10 and TGF-β contents were measured at the 14 th day. Regulatory T cells(Tregs)percentages were evaluated by flow cytometry.RESULTS:1.The sample was ultrasoniced extracted by 100 ml acetonitrile-0.4%phosphate(32:68) for 30 min.The TSK gel ODS-100Vcolumn(250 mm×4.6 mm,5 μm)was used and mobile phase was acetonitrile-0.4% phosphate(32:68) with the detection wavelength of 345 nm,the flow rate of 1 ml/min,and the column temperature.Palmatine and berberine contents in Ya-Jie-Sha-Ba were approximately 1.49 mg/g and0.50 mg/g.2.Scratching frequency, induced by OVA, was significantly suppressed in mice treated with either loratadine or palmatine or berberine, or 3.50, and 4.70 g/kg body weight Ya-Jie-Sha-Ba. OVA-sensitization increased serum Ig E amounts by 6.9% at 50 h,however, at 98 h, the serum Ig E amounts returned to baseline. Palmatine and Ya-Jie-Sha-Ba treatments reversed the OVA-induced increase in serum Ig E. The frequency of CD4~+CD25~+ Treg in the spleen increased from 6.80% in the control group to 12.50% in mice treated with 4.70 g/kg body weight Ya-Jie-Sha-Ba. Mice treated with palmatine or 4.70 g/kg body weight Ya-Jie-Sha-Ba had increased Foxp3 expression compared to those in the control group.3.Scratching frequency, induced by DNFB, was significantly suppressed in mice treated with the low dose of SFR and low and high dose of oxymatrine. The ear thickness was also reduced by SFR treatment. The frequency of CD4~+CD25~+ Treg in the spleen increased from 10.24% in the control group to 15.51% in high dose of oxymatrine group. High dose of SFR increased Foxp3 expression compared to those in the control group.CONCLUSIONS:1.Palmatine and berberine are considered as major active substances of Ya-Jie-Sha-Ba. This method was of high sensitivity, good repeatability, great stability and simple sample processing procedure.2.Ya-Jie-Sha-Ba decreased scratching frequency and increased CD4~+CD25~+Foxp3~+Treg frequency in the spleen indicating that the food allergic reaction was significantly alleviated following Ya-Jie-Sha-Ba treatment. Palmatine was identified as one of the major active ingredients of Ya-Jie-Sha-Ba. This research presented identified the possible mechanism through which Ya-Jie-Sha-Ba treatment alleviated food allergy symptoms in TDM.3. Sophorae Flavescentis Radix showed anti-allergic dermatitis effects via increasing CD4~+CD25~+Foxp3~+ Treg ratio. Oxymatrine was one of the major active ingredient of SFR.