| Background:Psoriasis is a chronic,inflammatory and recurrent skin disease with complex etiology.The pathogenesis of psoriasis has not been fully understood,and it is believed to be an immune response involving multiple immune cells mediated by T lymphocytes,.Since most patients reported obvious stress events when they first came on or relapsed,this disease was recognized as a mental and physical disease by the world health organization.At present,although systemic therapeutic drugs can eliminate the symptoms of diseases in clinical practice,they cannot completely cure diseases.However,traditional Chinese medicine(TCM)has a long history and has unique curative effect and minor toxic and side effects in the treatment of complex chronic diseases.According to the pathogenesis theory of TCM,we put forward the idea of treating psoriasis from liver,On the basis of the empirical prescription liangxue jiedu recipe commonly used in Beijing hospital of TCM,anti febrile liver meridian drugs were added to form the optimized prescription Qinggan Liangxue Jiedu recipe,which has good clinical efficacy.However,its effective components and mechanism are still unclear.Objective:Establish a psoriatic mouse model induced by anxiety combined with imiquimod.,to clarify the effect of psychological stress on psoriatic lesions in mice,and observe the effect of Qinggan Liangxue Jiedu recipe on the lesions of this model.use network pharmacology to explore the main components of Qinggan Liangxue Jiedu prescription and clarify formulas target and mechanism of skin lesions improvement.To provide a theoretical basis for the clinical application of Qinggan Liangxue Jiedu prescription in the treatment of psoriasis.Methods:Experiment 1:After empty bottles for stimulating two weeks,Imiquimod(IMQ)was used to build composite model.Behavioral assessment was made every week.The mice were sacrificed on the 4th.,7th,14th day,respectively.Take photos every day to observe dynamic change of model mice skin lesions,and obtain Psoriasis Area and Severity Index(PASI)score.Morphological changes of lesions were observed by HE staining.SP expression in skin lesions was detected by Immunohistochemical method.Expression of cytokines in skin lesions was detected by PCR.of neuropeptide expression in brain tissue and skin lesions were analyzed by Western Blot.Experiment 2:Observation the intervention effect of Qinggan Liangxue Jiedu decoction on compound model skin lesions.Changes of skin lesions and PASI score were obtained by HE staining.Morphological changes of the lesion were observed by HE staining and the thickness of the epidermis was measured.Proliferation Cell Nuclear Antigen(PCNA)was detected by immunohistochemistry and spleen index were measured.The proportion of CD11c/CD11b positive cells in spleen was detected by flow cytometry.Neuropeptide expression in brain tissue and skin lesions were analyzed by Western Blot.Experiment 3:The chemical molecular database of Qinggan Liangxue Jiedu detoxification prescription was constructed through literature mining and database collection.The ADME properties of drug molecules were evaluated by computer and the effective chemical components were obtained.The developed computer models are used to predict the direct targets of effective molecules.Drug-target-pathway network was constructed for network pharmacological analysis.Establish integrated pathway and reveal the mechanism of TCM compound through different pathway therapy modules.Results:Experiment 1:After empty bottles to stimulation,compared with the normal mice behavior,the mice in the Imiquimod induction of psoriasischanged significantly on skin(p<0.05).And field experiment showed that the movement percentage(Time and distance)in ES group were less than control group.Overhead cross maze experiments showed the open arms of time and frequency percentage in empty bottles stimulation group were less than the control group.Skin lesions related studies showed that the skin thickness increased significantly in the empty bottles and imiquimod stimulated group(p<0.01).And the PASI score and HE staining showed that skin lesions were aggravated.The results of PCR and WB experiments showed that the expressions of neuropeptide SP and inflammatory factors IL-1,IL-17A and IL-22 in the lesions were higher than those in the imiqumod model group(p<0.05),indicating that empty bottle stimulation had potential aggravating effect on lesions.Experiment 2:According to PASI scoring and HE staining of paraffin sections of skin lesions,the high,medium and low dose group of Qinggan Liangxue Jiedu prescription showed improvement in the skin lesions of the compound model group and the pure imiquimod model group,the thickness of the epidermis also decreased.Qinggan Liangxue Jiedu decoction can reduce the spleen index and the number of CD11c+/CD11b+ positive cells,with dose-dependent effect.Immunohistochemical results showed that Qinggan Liangxue Jiedu recipe could inhibit the number of PCNA positive cells in mouse skin.In addition,Qinggan Liangxue Jiedu recipe can significantly reduce the expression level of SP and NK-IR in the skin,which indicated it maybe play an anti-inflammatory roleExperiment 3:The effective compounds in Qinggan Liangxue Jiedu detoxification prescription were collected to establish a database.Then drug targets were screened and the drug-target,target-disease,target-pathway and disease-disease interaction network were established.Through GOBP analysis of drug-target,the first 23 enriched GOBP entries that are closely related to the potential target of Qinggan Liangxue Jiedu recipe and psoriasis were listed.Many targets are closely related to the biological process of psoriasis.Such as inflammatory reaction,positive regulation of NF-kappa B transcription factor activity,immune response,positive regulation of MAP kinase,positive regulation of interleukin-6 production and so on.Cytoscape software was used to visualize the activation or inhibition relationship between a compound and its target.Finally,a chemical-target interaction network consisting of 315 nodes(151 compounds and 164 potential targets)and 1095 edges was obtained.In order to explore the biological effects of drugs at the pathway level,we constructed a "psoriasis disease pathway" through literature mining and sorting.The results showed that the targets of Qinggan Liangxue Jiedu recipe were mainly involved in four major signaling pathways,including MAPK signaling pathway,T cell receptor signaling pathway,toll-like receptor signaling pathway and NF-κB signaling pathway.We constructed the pattern of signal pathway of Qinggan Liangxue Jiedu prescription in the treatment of psoriasis,and a large number of targets were involved in the interaction of the above four signal pathways,such as TLR4,TNFR,TGFBR,mekl/2,ERK,etc.Conclusion:Empty bottle stimulation can aggravate the skin lesions of mice psoriasis induced by imiqumod,while Qinggan Liangxue Jiedu recipe can reduce the skin lesions of compound model mice,reduce epidermal thickness,and inhibit the expression of inflammatory factors and neuropeptide SP.The specific mechanism may involve NK-1R receptor and related signals.In addition,Qinggan Liangxue Jiedu recipe is a multi-component and multi-target drug. |
