SERPINB3/4 Overexpression Promotes Rosacea-and Psoriasis-like Skin Inflammation By Activating NF-κB Signaling

Background: Rosacea and psoriasis are common chronic inflammatory skin diseases with a high incidence,which are prone to recurrent clinical attacks and bring great pain to patients.Although previous studies have suggested that abnormal communication between keratinocytes and immune cells plays an important role in both pathogenesis,the common molecular mechanism of keratinocytes and immune cells is still lacking.Exploring the common molecular basis of these diseases will contribute to the development of new antiinflammatory targeted drugs.In the previous study,transcriptome sequencing analysis showed that the expression levels of Serine Protease Inhibitor(SERPIN)family B subgroups 3 and 4(SERPINB3/4)were significantly increased in the skin lesions of rosacea patients.At the same time,we analyzed transcriptome sequencing data of psoriatic skin lesions in previous studies,and found that SERPINB3/4 expression level in psoriatic skin lesions was also significantly increased.Therefore,we focused on the specific role and molecular mechanism of SERPINB3/4 in the pathogenesis of two inflammatory skin diseases.Objective: In this study,our primary objective is to further clarify the common molecular mechanisms underlying the pathogenesis of rosacea and psoriasis,identify the specific roles of SERPINB3/4 in both diseases,and finally find out the potential therapeutic targets to develop new therapies.Methods: In this study,the expression of SERPINB3/4 in serum and skin lesions of patients with rosacea and psoriasis and the expression of Serpinb3a(a mouse homologue of SERPINB3/4)in LL37-induced rosacea-like mouse model and imiquimote induced psoriasis-like mouse model were determined by means of ELISA,immunohistochemistry,q PCR and immunofluorescence.Then,we used small interfering RNA,si RNA intradermal injection to locally knock down the expression of Serpinb3 a in mouse skin,and observed its effect on the inflammatory manifestations and skin histopathology in the inflammatory model.In vitro studies were conducted mainly in Ha Ca T cells(human immortal keratinocytes).SERPINB3/4 was overexpressed and knocked down by plasmid transfection or lentivirus infection in human Ha Ca T cells.RNAseq,western blotting,and immunofluorescence were used to determine its relationship with NF-κB signaling pathway.Meanwhile,we selected SC75741(a potent inhibitor of NF-κB signaling pathway)to inhibit NF-κB signaling pathway,and to explore the role of NF-κB signaling pathway in the pathogenesis of rosacea.Finally,the regulation mechanism of SERPINB3/4 in promoting the pathogenesis of rosacea and psoriasis through NF-κB signaling pathway/chemokine signaling axis was further identified in vitro cell experiments through gene overexpression,bioinformatics analysis,q PCR and other techniques.Results:(1)The serum level of SERPINB3/4 was significantly increased in rosacea patients,and the expression level was positively correlated with the severity of the disease.(2)SERPINB3/4 expression was increased in rosacea and psoriasis lesions;(3)Knocking down Serpinb3 a improved the inflammatory phenotype of rosacea-like and psoriasis-like mice;(4)SERPINB3/4 in human keratinocytes positively regulated NF-κB signaling pathway;(5)Inhibition of NF-κB signaling pathway alleviates the skin lesion phenotype of rosacea.(6)SERPINB3/4 induced diseaserelated chemokines and cytokines through activation of NF-κB signaling pathway,and further promotes T cell chemotaxis.Conclusions: First of all,the expression of SERPINB3/4 was significantly increased in rosacea and psoriasis patients,and its expression level was positively correlated with the severity of the two disease.It is preliminarily believed that SERPINB3/4 can be used as an indicator of the diagnosis and prognosis of the two skin diseases in clinic.In addition,SC75741,a potent inhibitor of NF-κB signaling pathway,significantly improved the inflammatory phenotype of rosacea-like mouse skin lesions,and inhibited the activation of NF-κB signaling pathway induced by SERPINB3/4 overexpression in human Ha Ca T cells in vitro,providing a new therapeutic idea for the treatment of rosacea and psoriasis.Finally,this study reveals the key role of SERPINB3/4 in the abnormal dialogue between keratinocytes and immune cells during the development of rosacea and psoriasis,namely,the overexpression of SERPINB3/4 in keratinocytes releases inflammatory cytokines and chemokines through activation of the NF-κB signaling pathway,which chemotaxis T cells.They interact with each other and eventually promote the occurrence of inflammation in the lesions of rosacea and psoriasis.