| There has been a global increase in the incidence rate of allergic inflammatory skin disorders.In clinical practice,diseases related to typeⅣallergy,like contact dermatitis and eczema etc,are quite frequently found in patients with chief complaints of skin problems. Pathomechanism of these diseases involves expression of several important cytokines,which participate in hypersensitive responses.Currently,the most emphasized ones are interferon-γ(IFN-γ),tumor necrosis factor-α(TNF-α),intercellular adhesion molecule-1(ICAM-1)and vascular cell adhesion molecule-1(VCAM-1).Xi Jiao Di Huang Tang(Cornus Rhinoceri and Rehmanniae Decoction, CRRD),originated from Bei Ji Qian Jin Yao Fang(Essential Prescriptions Worth a Thousand Gold for Emergencies),has the functions of cooling blood and dissipating stasis,clearing heat and eliminating toxicity.Through long-term clinical application and testing,it is generally acknowledged as a representative formula for Xue level febrile diseases.Clinical reports of treating dermatological disorders with this formula have been numerous with satisfactory result.Since Xi Jiao(Rhinoceros Horn)is a rare and protected medicinal,Shui Niu Jiao(buffalo horn)is often used in clinic replacing Xi Jiao with a fairly similar therapeutic effect.This formula can be modified to effectively treat various allergic skin disorders diagnosed as blood heat pattern in Chinese Medicine, including diseases like drug rash,eczema,and contact dermatitis etc. Previous researches showed CRRD has antipyretic,sedative,anticonvulsion, anti-inflammatory,antibiotic and immune-boosting functions.To explore the mechanism of CRRD in treating allergic skin disorders, this study observed the anti-allergic and anti-inflammatory effects of CRRD(buffalo horn)in various approaches.Two disease models of allergic contact dermatitis(induced by DNCB)and photoallergic contact dermatitis (provocated by UVA and induced by 6-MC)were used in mice.The effects of CRRD(buffalo horn)on disease models including sizes of swollen ears, thymus and spleen index were observed.Levels of IFN-γand TNF-αwere also measured using double antibody ABC-ELISA and radioimmunity analysis (RIA).The changes of IFN-γand TNF-αwere measured using RT-polymerase chain reaction(RT-PCR)method.The expressions of ICAM-1 and VCAM-1, pathological changes of local inflammation were observed by immuno- histochemical methods.Therefore the anti-inflammation and anti-allergy actions of CRRD(buffalo horn)were studied in various approaches: pathology,cytology and molecular biology.The actionmechanism and target of CRRD(buffalo horn)in treating allergic skin disorders were explored, which might be useful in promoting the clinical application of traditional Chinese medicine formula.Results:1.Large dose of CRRD(buffalo horn)could significantly inhibit the swelling of the ears in mice induced by DNCB,compared with model control group(p＜0.05).Besides,spleen index in CRRD(buffalo horn) group was also significantly reduced(p＜0.05).Thymus index was significantly lowered in model control group while hydrocortisone group as a positive control,the lowest.Yet CRRD(buffalo horn)groups shared a similar thymus index with model control group.2.IFN-γin mice serum was significantly increased in model group(allergic contact dermatitis induced by DNCB)and CRRD(buffalo horn)could inhibit this increase by a significant difference compared with model control group(p＜0.05).Low dose of CRRD(buffalo horn)didn't show such action.Meanwhile hydrocortisone could also inhibit the increase of IFN-γwith a significant difference(p＜0.05).3.Sizes of the swelling ears induced by 6-MC was significantly different from healthy control ones(p＜0.01) at 24h,48h and 72h.Both CRRD(buffalo horn)and hydrocortisone could significantly inhibit the swelling of the ears and spleen index induced by 6-MC(compared with mice model control group,p＜0.05).There is insignificant difference in thymus index between model control group and healthy control group.In hydrocortisone group,thymus index decreased significantly at 24h and 72h(p＜0.01),especially at 48h(p＜0.001) compared with healthy control group.4.At 24h after induced by 6-MC,There was no change of serum IFN-γlevel in model control group and no difference between healthy and model group.At 48h,IFN-γincreased significantly in model control group compared with healthy control group (p＜0.05).Both CRRD(buffalo horn)and hydrocortisone could inhibit the increase of IFN-γsignificantly compared with healthy control group (p＜0.05).5.At 24h after induced by 6-MC,TNF-αincreased significantly in model control group compared with healthy control group(p＜0.01).This significant increase continued at 48h(p＜0.05).Both CRRD(buffalo horn) and hydrocortisone can inhibit this increase at 24h and 48h with a significant difference from model control group(p＜0.05).6.Expression of both ICAM-1 and VCAM-1 increased significantly in model control group (p＜0.001).CRRD(buffalo horn)can restrain this increase.Although the expression of ICAM-1 and VCAM-1 positive cells in CRRD(buffalo horn) group was significantly higher than healthy control group,it was still significantly different from model control group(p＜0.01).Hydrocortisone can also inhibit the increased expression significantly compared with model control group(p＜0.01)The result indicates that:1.CRRD(buffalo horn)can significantly inhibit the latent allergic dermatitis(induced by DNCB)and photoallergic contact dermatitis(provocated by UVA and induced by 6-MC), which provides research evidence for clinical application of this formula; 2.CRRD(buffalo horn)can inhibit the increase of IFN-γand TNF-αwith a similar effect to hydrocortisone,which indicates a possible mechanism of inhibiting inflammatory cytokines in the formula's anti-inflammation and anti-allergy function;3.CRRD(buffalo horn)can counterfeit the increased expression of ICAM-1 and VCAM-1,which indicates CRRD(buffalo horn)inhibit inflammation by inhibiting the expression of adhesion molecules in the skin lesions of allergic dermatitis model mice;4.CRRD (buffalo horn)doesn't affect the weight of immune organs in model mice, nor its total weight,which shows no apparent harm to the immune organs of CRRD(buffalo horn)while it can inhibit immune responses.
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