| Chapter ?ALA-PDT alleviates the psoriasis-like lesions in the imiquimod-induced miceBackground:Psoriasis is a chronic and inflammatory systemic skin disease,which affects approximately 2-3%of the general population around the world.The primary cause of the disease leading to abnormal differentiation and impaired barrier function,was the hyperproliferation of epidermal keratinocyte.Many types of PDT have been developed and used for the treatment of psoriasis over the last few decades.IMQ induced psoriasis is a widely used method for creating psoriasis pathological mouse model for in vivo studies.Objectives:In this study,to investigate ALA-PDT treatment effectively ameliorated the psoriasis-like lesion in imiquimod(IMQ)-induced mouse model and further explored the potential molecular mechanism and related signaling pathways during the treatment.Methods:Use seven-week-old female BALB/c mice undergoing imiquimod induction as the psoriasis model.Five mice in the first group did not receive any treatment and served as a control.Five mice in the second group received a topical treatment of imiquimod on their back.Five mice in the third group received imiquimod combined with red light treatment.The mice in the fourth group received ALA-PDT followed by imiquimod.Five mice in the fifth group received imiquimod combined with ALA-PDT.To investigate the pathological index,hematoxylin and eosin(H&E)staining,the levels of proteins(K17,JAK pathway)Results:1.1 ALA-PDT suppresses imiquimod-induced psoriasis-like lesionsALA-PDT treatment effectively ameliorated these symptoms,characterized by the nearly disappeared reddening and reduced scaling skin in IMQ-induced mice.The erythema,scales,thickness and cumulative scores of the IMQ-treated area were significantly decreased with the treatment of ALA-PDT.1.2 ALA-PDT suppresses imiquimod-induced psoriasis-like lesions hyperkeratosis After treating with ALA-PDT,the degree of keratosis was significantly reduced,granular layer cells were enriched,the arrangement of different cells was returned to the normal levels.Repeated treatment with ALA-PDT has the greater therapeutic efficacy.Only treated with ALA or irradiation has not changed the histological features of IMQ-induced skin.1.3 ALA-PDT prevented imiquimod-induced the expression of K17 by the JAK pathwayAfter treating with ALA-PDT,K17 expression level was significant decreased.Both JAK1 and JAK2 protein were over-expressed in IMQ-induced dorsal skin as compared to the normal dorsal skin.After treating with ALA-PDT,JAK1 and JAK2 expression levels were largely reduced.Expression of Socs1 and Socs3 protein were up-regulated in IMQ-induced dorsal skin,treated with ALA-PDT further up-regulated their expression.Conclusion:ALA-PDT alleviates the psoriasis-like lesions in the IMQ-induced mice and also promotes the SOCs1/3 and inhibits the JAK 1/2 expressions on the psoriasis-like skin in IMQ-induced miceChapter ?ALA-PDT inhibits the cell proliferation in IFN-?-induced keratinocytes by JAK pathwayBackground:Earlier studies have shown that interferon(IFN)-? produced by natural killer(NK)cells and CD4+T cells has pro-inflammatory effects on keratinocytes,and plays a central role in the overall pathogenesis of psoriasis.Also,the IFN-y level in serum is correlated with clinical severity and activity of psoriasis.Thus,IFN-y might be an inducer of keratinocyte hyperproliferation,and K17 could serve as a marker for evaluating the effect of psoriasis therapy.Recently,JAK proteins have emerged as possible new therapeutic targets for the treatment of autoimmune inflammatory diseases,including psoriasis.The JAK/signal transducers and activators of transcription(STAT)pathway plays a critical role in mediating inflammatory immune responses by converting cytokine signals into genomic responses regulating proliferation and differentiation of the immune cells.At molecular level,SOCs1 and SOCs3,as members of suppressors of cytokine signaling(SOCs)genes,which are well known negative feedback regulators,have effect on inhibiting JAK 1/2 by functioning as pseudo-substrates.Objective:In this study,to investigate ALA-PDT treatment effectively ameliorated the psoriasis-like lesion in IFN-y-induced keratinocytes model and further explored the potential molecular mechanism and related signaling pathways during the treatment.Methods:Use keratinocytes undergoing IFN-y induction as the psoriasis model.Keratinocytes were divided into following four groups:NC,ALA-PDT,IFN-?,IFN-?+ALA-PDT.The first group did not receive any treatment and served as a control.The second group received a topical treatment of IFN-y.The third group received only ALA-PDT.The fourth group received ALA-PDT followed by IFN-y.To investigate the cell viability,cell cycle,the levels of proteins(K17,JAK pathway).Results:1.1 ALA-PDT inhibits the cell proliferation in IFN-y-induced keratinocytesThe proliferation ability of keratinocytes significantly inhibited by exposing to IFN-y,but not influenced with ALA-PDT treatment.Interestingly,after treating with ALA-PDT,the proliferation of IFN-? treated keratinocytes was decreased.Correspondingly,cell cycle was analyzed by flow cytometry,the proportion of G1 phase in cell cycle was higher and G2 phase was lower in the IFN-y treated keratinocytes than normal keratinocytes.Treatment with ALA-PDT effectively increased the proportion of G1 phase and reduced the proportion of G2 phase of the IFN-? treated keratinocytes.1.2 ALA-PDT prevented IFN-y-induced keratinocytes expression of K17 by the JAK pathwayAfter treating with ALA-PDT,K17 expression level was significant decreased.Both JAK1 and JAK2 protein were over-expressed in IFN-?-induced keratinocytes as compared to the normal.After treating with ALA-PDT,JAK1 and JAK2 expression levels were largely reduced.Expression of Socsl and Socs3 protein were up-regulated in IFN-y-induced keratinocytes,treated with ALA-PDT further up-regulated their expression.Conclusion:ALA-PDT alleviates the IFN-y-induced keratinocytes and also promotes the SOCsl/3 and inhibits the JAK 1/2 expressions on the psoriasis-like IFN-y-induced keratinocytesChapter ?ALA-PDT attenuates the proliferation of IFN-y-induced keratinocytes by promoting reactive oxygen species(ROS)levelBackground:Photodynamic therapy(PDT),also known as oxygen-dependent photochemical reaction,occurs upon a photochemical reaction between light,photosensitizer(PS)and molecular oxygen,leading to directly induce cellular damage and accelerate cells death.Many types of PDT have been developed and used for the treatment of psoriasis over the last few decades.The outcome of phototherapy depends on a delicate balance between beneficial and detrimental effects of a specific laser,but its side effects limit its broad application.Phototherapy can be combined with biologic agents for the treatment of severe psoriasisObjective:In this study,to investigate ALA-PDT treatment effectively ameliorated the psoriasis-like lesion in IFN-y-induced keratinocytes model and further explored the potential molecular mechanism and related ROS during the treatmentMethods:Keratinocytes were divided into following four groups:IFN-?,IFN-?+ALA-PDT,IFN-y+NAC,.IFN-y+ALA-PDT+NAC.The first group received a topical treatment of IFN-y as a model.The second group received a topical treatment of ALA-PDT.The third group received IFN-? combined with NAC.The fourth group received ALA-PDT followed by NAC.To investigate the cell viability,cell cycle,the levels of proteins(K17,JAK pathway).Results:1.1 ALA-PDT attenuates the proliferation of IFN-y-induced keratinocytes by promoting reactive oxygen species(ROS)levelThe proliferation ability of IFN-y treated keratinocytes was significantly attenuated with ALA-PDT treatment and the proliferation ability of IFN-? treated keratinocytes was significantly enhanced with NAC treatment.By contrast,this markedly down-regulated change on proliferation in ALA-PDT treated keratinocytes was restricted when co-treated with ALA-PDT and NAC.Correspondingly,the proportion of G1 phase in the cell cycle of IFN-y treated keratinocytes was increased and G2 phase was decreased after treating with ALA-PDT.Treatment with NAC effectively decreased the proportion of G1 phase and increased the proportion of G2 phase of the IFN-y treated keratinocytes.1.2 ALA-PDT attenuates the expression of K17 by the JAK pathway by promoting reactive oxygen species(ROS)levelExpression of K17 protein in IFN-?-exposed keratinocytes was significantly down-regulated with ALA-PDT treatment,co-treated with ALA-PDT and NAC reversed this change.The JAK1 and JAK2 protein exhibited the similar changes with K17.Additionally,both Socsl and Socs3 were obviously up-regulated in IFN-y treated keratinocytes after ALA-PDT treatment.When co-treated with ALA-PDT and NAC in IFN-? treated keratinocytes,their expressions down-regulated again.Conclusion:ALA-PDT alleviates the IFN-y-induced keratinocytes and also promotes the SOCsl/3 and inhibits the JAK1/2 expressions by promoting ROS level. |
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