Anti-psoriasis Activity And The Action Mechanism Of Danshen Baibixiao Pills

Psoriasis is a chronic immune-mediated inflammatory disease.Psoriasis is of high morbidity and difficult to cure,which increase the burden on patients’ health physically and psychologically.There are several disadvantages with the currently used medicines,such as side effects,inconvenience to use and expensive price,which limit the use of these medicines.Traditional Chinese medicine could be an important way to treat psoriasis.Danshen Baibixiao Pills(DB)have been efficiently used to treat psoriasis for more than 20 years,but the underlying mechanisms have never been investigated.This study used in vitro and in vivo models to evaluate the anti-psoriasis activity of DB,investigated its underlying mechanisms,and analyzed the active constituents of DB.Objective: To evaluate the anti-psoriasis activity of DB with in vitro models and imiquimod(IMQ)-induced psoriasis-like mice models and investigate its underlying mechanisms.To analyze the active constituents of DB with HPLC-MS/MS methods.Methods: 1,IMQ-induced psoriasis-like mice models were used to affirm its efficacy against psoriasis.The psoriasis area severity index(PASI)was assessed,H&E staining was performed,and the epidermal thickness was measured.The levels of tumor necrosis factorα(TNF-α),interleukin(IL)-17 A,IL-23,IL-6,IL-1β and IL-22 in serum were assessed by ELISA.The mRNA expressions of skin IL-17 A,IL-23,IL-6 and IL-22 were determined by real-time polymerase chain reaction(PCR).Expression levels of proteins related to NF-κB,MAPKs and STAT3 signaling pathways were mesured by western blotting;2,the anti-proliferation effect of DB on human keratinocytes,HaCaT cells was evaluated with MTT methods.3,The effects of DB on the secretion levels of NO,TNF-αand IL-1β of LPS induced RAW264.7 cells were investigated by ELISA.4,six constituents,salvianolic acid B,tanshinone IIA,berberine hydrochloride,cimicifugin,sophocarpidine,and ammonium glycyrrhetate were quantified using HPLC-MS/MS with multiple reaction monitoring(MRM)mode.Results: 1,Oral administration of DB ameliorated the IMQ-induced psoriasis-like dermatitis.IMQ-induced keratinocytes’ proliferation was alleviated,and the epidermal thickness decreased.The serum levels of inflammatory cytokines(IL-17,IL-23,IL-6,IL-22,TNF-αand IL-1β)declined,the mRNA expressions of IL-17 A,IL-23,IL-6 and IL-22 in skin tissues were down-regulated.The expression level of IκB was increased and the expressions of NF-κB p65,p38,JNK,ERK,STAT3,p-p38 and p-ERK were decreased significantly after DB treatment.2,DB showed significant anti-proliferation effect on HaCaT cells.3,DB extract significantly declined the secretion levels of NO,TNF-αand IL-1β of LPS induced RAW264.7 cells.The aqueous extract of DB showed better activity than the alcohol extract of DB.4,The concentrations of salvianolic acid B,tanshinone IIA,berberine hydrochloride,cimicifugin,sophocarpidine,and ammonium glycyrrhetate in DB extract were 12.3,0.13,6.71,3.04,49.5,5.96 μg/mg.Conclusions: DB showed activity to inhibit the proliferation of HaCaT cells and the productions of inflammatory cytokines by LPS induced RAW264.7 cells,which suggest that the anti-psoriasis activity of DB is related with the activity to inhibit the proliferation of keratinocytes and the productions of related inflammatory cytokines.DB showed obvious anti-psoriasis activity in the IMQ-induced psoriasis-like mice model.Its mechanism was related to the activities of regulating the IL-23/IL-17 axis and suppressing the activation of NF-κB,MAPKs and STAT3 signaling pathways.These results suggested that DB could be a potential alternative therapy for psoriasis treatment and provided the theory basis for the clinical application of DB.