UPLC-Q-TOF/MS And Network Pharmacology-based Study On The Mechanism Of Qingre Lishi Decoction In Psoriasis Therapy

BackgroundPsoriasis is a common immune mediated chronic recurrent inflammatory systemic disease in clinical practice,characterized by erythema and scaling,often affecting joints,and can also be accompanied by metabolic syndrome,cardiovascular disease,anxiety,depression,and other diseases.The pathogenesis of psoriasis is complex,and its treatment is particularly important due to its stubbornness.Currently,the main medications for psoriasis treatment include methotrexate,acitretin,cyclosporine,and biological small molecule targeted drugs,especially biological agents targeting the IL-23/Th17 axis,which have definite therapeutic effects.Although these medications have a quick onset of action,they also have various problems,such as systemic immunosuppression,immunological imbalance,high relapse rate after discontinuation,high cost,heavy burden on medical insurance,limitations for patients with infections or tumors,etc.Traditional Chinese medicine(TCM)has recorded and explained the etiology,pathogenesis,differentiation and treatment of psoriasis.As one of the most commonly used treatment methods for psoriasis,TCM has definite clinical efficacy and minimal toxic side effects.Qingre Lishi Decoction is a classic formula of Qilu Du’s dermatology school,which has been used for the treatment of psoriasis for more than 40 years.However,the lack of effective ingredients and mechanism research has limited its clinical promotion and application.Therefore,clarifying the effective active ingredients of Qingre Lishi Decoction,predicting its main targets of action and signal pathways,and selecting TCM formulas with consistent principles and formulas based on symptoms for mechanism research are important topics to address clinical needs and serve as a bridge between traditional Chinese medicine and modern clinical research.ObjectiveTo clarify the active ingredients of Qingre Lishi Decoction,predict its main targets and signaling pathways,and verify its mechanism of action against psoriasis through in vitro experiments.MethodsUltra performance liquid chromatography-quadrupole-time of flight-mass spectrometry(UPLC-Q-TOF/MS)technology was used to identify the components of Qingre Lishi Decoction and its blood components.Natural product databases were used to obtain the target proteins of the blood components of Qingre Lishi Decoction and those related to psoriasis.A"drug-component" network and a target protein-related interaction network were constructed.GO and KEGG pathway enrichment analysis was performed using the DAVID database.Molecular docking was performed between the key active ingredients and core target proteins to further verify the binding affinity of the ingredients and targets.HaCaT cells were used as a psoriasis cell model,and Qingre Lishi Decoction-containing serum and core target protein-related factors were used for intervention.CCK-8 was used to detect the proliferation of cells treated with different concentrations of serum,and RT-PCR and immunofluorescence were used to verify the changes in inflammatory pathways mediated by target proteins.Results1.Based on UPLC-Q-TOF/MS technology,57 chemical components were identified from the compound of Qingre Lishi Decoction,and 15 prototype components and 22 metabolites were identified in the plasma samples.2.Through network analysis,23 intersecting genes were obtained from 260 component-related drug targets and 218 psoriasis-related disease targets.TNF,IL-10,STAT3,LGALS3 and other potential key core targets for the treatment of psoriasis were identified.GO and KEGG enrichment analysis showed that key targets were mainly involved in the regulation of cytokine-mediated signal transduction pathways,cell proliferation and migration,as well as pathways related to inflammatory diseases and immune-related diseases,such as TNF-mediated signaling pathways and T cell receptor signaling pathways.Molecular docking results showed that multiple active ingredients in Qingre Lishi Decoction can bind well to the target proteins TNF and LGALS3 of psoriasis(binding energy<-5 Kcal/mol).3.In vitro studies found that Qingre Lishi Decoction can inhibit ITG(10 ng/ml IL-17A,10 ng/ml TNF-α and 100 ng/ml Gal-3)induced proliferation of HaCaT cells,and may alleviate inflammatory reactions by targeting TNF.Compared with the model group,the treatment group showed a significant decrease in the expression of IL-6,IL-1β,and IL-17,and an increase in the expression of IL-10(P<0.05).Qingre Lishi Decoction-containing serum also effectively inhibited TNF-α-induced p65 nuclear translocation.ConclusionThis study preliminarily revealed the active ingredients and key targets of Qingre Lishi Decoction for the treatment of psoriasis,and its potential mechanism of action against psoriasis through inhibiting TNF-α-mediated inflammatory reactions.This provides a theoretical basis for the clinical promotion and application of Qingre Lishi Decoction in the treatment of psoriasis.