The Effect And Mechanism Of Using Sophora Flavescens And Rhizoma Smilacis Glabrae Together To Alleviate Psoriasis-like Skin Lesions In Mice

Rationales:Psoriasis is a chronic and refractory skin disease that can cause systemic diseases and seriously endanger the physical and mental health of patients.The number of patients is increasing year by year.Psoriasis currently requires lifelong treatment.The primary purpose of treatment is to control symptoms and improve the quality of life of patients.Existing drugs cannot meet the needs of patients.Rapid removal of skin lesions and prevention and treatment of recurrence are two unresolved scientific problems about the treatment of psoriasis.Using traditional Chinese medicine treatment of psoriasis has great advantages.Sophora flavescens and smilax glabra are common Chinese herbal medicines for the treatment of psoriasis,so the combination of the two is more feasible for the treatment of psoriasis.This study explores the effect and mechanism of Sophora flavescens and Smilax glabra in improving psoriasis,which is of great significance for the development of new drugs for psoriasis.Aim: To explore whether SF and SRG synergistically improve psoriasis-like skin lesions in mice and the effect on disease recurrence after healing,and to preliminarily analyze its mechanism of action.Expecting to provide a new drug combination for the treatment of psoriasis.Methods:1.The effects of SF and RSG alone-use and in combination on psoriasis-like lesions in mice were studied by using imiquimod-induced mouse models of psoriasis in progressive and recurrent stages,PASI score,HE staining and etc.And the recurrence of the disease was further evaluated.2.Immunohistochemistry and western Blot were used to detect the levels of the proliferation and differentiation of keratinocytes and T cell related marker proteins in psoriatic lesions of mice,and to evaluate the effect of SF combined with RSG on the positive feedback loop of ’ keratinocytes-immune microenvironment ’ in psoriasis of mice.3.The effects of SF combined with RSG on the growth of keratinocytes(Ha Cat)cells and the expression of related pathway genes were verified at the cellular level.The effect of SF combined with RSG on the activity of Ha Cat cells was studied by sulforhodamine B staining(SRB),and the inhibitory effect of SF combined with RSG on Ha Cat cells was observed.The expression of STAT3 gene was determined by constructing Ha Cat cells containing Stat3 gene and then detecting the luciferase reporter gene.4.UPLC-Triple TOF 5600 was used for metabolomics analysis of serum,HMDB database,MSDAIL software and SCIXE OS software were used to analyze the data,SIMCA software was used to find the differential metabolites in serum,to find out the key pathway of SF combined with RSG treatment of psoriasis-like lesions in mice.Result:1.The back pictures of mice showed that the combination of SF and RSG and the positive drug(Vtama)could reduce the skin erythema and scales of psoriasis in mice.The results of PASI score showed that the combination of SF and RSG could reduce the PASI score of psoriasis in mice.HE staining was used to evaluate the skin structure of skin lesions in mice,indicating that the combination of SF and RSG can reduce the epidermal thickness of psoriasis in mice.SF and RSG alone had no significant effect on psoriasis lesions.To further evaluate the recurrence of psoriasis after cure,the results showed that the combination of SF and RSG effectively reduced the recurrence of psoriasis in mice after cure,and the efficacy was comparable to that of Vtama.In summary,the combination of SF and RSG improves psoriasis-like lesions in mice and alleviates their recurrence.2.Immunohistochemistry was used to detect cell proliferation marker Ki67 and T cell marker CD3.The results showed that the combination of SF and RSG significantly reduced the expression of Ki67 and CD3 in psoriasis lesions of mice during treatment and recurrent stage after treatment,and the inhibition effect on the expression of Ki67 and CD3 in recurrent lesions after treatment is better.Western blot results showed that the combination of SF and RSG significantly reduced the expression of keratin 5(KRT5)and keratin 14(KRT14),the key proteins of psoriasis.In vitro cell level experiments results showed that the combination of SF and RSG significantly inhibited the growth of Ha Cat cells and inhibited the activation of psoriasis-related inflammatory pathway gene STAT3 in Ha Cat cells.In summary,the combination of SF and RSG inhibited the proliferation of keratinocytes in psoriasis lesions,promoted their normal differentiation,and reduced the number of T cells in lesions.3.The levels of key proteins H3K27 ac and H3K4Me1 in psoriasis recurrence were detected by immunohistochemistry.The results showed that the combination of SF and RSG significantly regulated the levels of H3K27 ac and H3K4Me1 in psoriasis lesions of mice after treatment.LC-MS metabolomics analysis results showed that the combination of SF and RSG significantly improved the metabolic abnormalities of psoriasis mice.The significantly regulated metabolic pathways were: arachidonic acid metabolism,sphingolipid metabolism,tryptophan metabolism,phenylalanine metabolism.In summary,the potential mechanism of the combination of SF and RSG to slow down the recurrence of psoriasis is to regulate histone epigenetic modification and metabolic processes.Conclusion: The combination of SF and RSG improves psoriasis-like lesions in mice by inhibiting the proliferation of epidermal keratinocytes and reducing the number of T cells in the skin,and delays the recurrence of psoriasis by regulating histone epigenetic modification and metabolic processes.The completion of the project provides a drug combination for the development of therapeutic drugs to slow the recurrence of psoriasis.