Shikonin Inhibits The Expression Of VEGFA In Psoriatic Keratinocytes Through NLRC4 Inflammasome

Objective: Psoriasis is a chronic,relapsing,lifelong inflammatory system disease caused by abnormal immune response,which is affected by the combined effects of heredity and environment.Numerous studies have shown that skin lesions in patients with psoriasis occur throughout the body,resulting in difficulty sleeping and a decrease in quality of life.The risk of hyperlipidemia,type 2 diabetes mellitus,coronary artery disease is greatly increased,and there is a greater risk of depression.At present,there is no safer and more efficient treatment for psoriasis.Shikonin is an anthraquinone derivative extracted from the traditional Chinese herbal medicine,which has antioxidant,anti-inflammatory,antithrombotic and wound healing effects,and has been used in the treatment of various inflammatory diseases.NLRC4 belongs to the NLR family,and protein members of this protein family play crucial roles in pathogen recognition and regulation of innate immune responses.This study mainly explores that shikonin affects the occurrence and progression of psoriasis by affecting the expression of NLRC4 and regulating the expression of Caspase-1,IL-1β and VEGFA.This study explored the role of shikonin in the treatment of psoriasis,and clarified the regulatory effect of shikonin on NLRC4 and the molecular mechanism of the effect of NLRC4 on the expression of Caspase-1,IL-1βand VEGFA.Targeted therapy provides new ideas in the treatment of psoriasis.Research methods: In this study,we used the keyword "psoriasis" in the GEO database to conduct a dataset search of Microarray data in the skin lesions and non-lesion areas of psoriasis patients.Bioinformatic prediction of differential gene comparisons in psoriasis patient and normal skin by Venn diagrams and volcano plots.In this study,Ha Ca T keratinocytes were selected as the evaluation cells for evaluating the therapeutic effect of psoriasis and IMQ-induced mice were used as the in vivo animal experimental model for this study.The effects of shikonin at gradient concentrations on the proliferation of Ha Ca T keratinocytes were detected by CCK8 method.Detection of inflammatory stimulation by western blot the expression changes of NLRC4 in Ha Ca T keratinocytes;the expression changes of NLRC4,Caspase-1,IL-1β and VEGFA proteins in Ha Ca T keratinocytes with gradient concentrations of shikonin after inflammatory stimulation;after overexpression of NLRC4,Caspase-1,IL-1β and VEGFA protein expression changes;after different doses of shikonin treatment,IMQ-induced changes in the expression of NLRC4,Caspase-1,IL-1β and VEGFA protein in mouse skin tissue.NLRC4 was overexpressed in Ha Ca T keratinocytes by lentiviral transfection.The transfection efficiency of NLRC4 was detected by RT-q PCR;after overexpression of NLRC4,the expression of Caspase-1,IL-1β and VEGFA m RNA changed.Results: In this study,bioinformatics prediction of multiple databases was firstly performed.The results confirmed that the gene expression in psoriasis patients and normal human skin tissues were differentially analyzed and compared.There were 597 common differential genes,and they were in GSE14905,GSE30999,GSE50790.and GSE53552 datasets NLRC4 is highly expressed in psoriatic tissues.This study determined the effect of shikonin on Ha Ca T keratinocytes.The expression of NLRC4 was significantly increased in Ha Ca T keratinocytes after inflammatory stimulation.By treating Ha Ca T keratinocytes with gradient concentrations of shikonin,NLRC4 could significantly reduced Caspase-1/IL-1β/VEGFA expression level.In this study,NLRC4 was further overexpressed in Ha Ca T keratinocytes,and it was found that after overexpression of NLRC4,the expressions of Caspase-1,IL-1β and VEGFA were significantly increased,confirming that NLRC4 affects the expression of Caspase-1,IL-1β and VEGFA by affecting the expression the development of psoriasis.Through IMQinduced psoriasis mouse model,treatment with different doses of shikonin can improve the skin lesions of IMQ-induced psoriasis mice and increase epidermal thickness.Through further molecular mechanism exploration,we found that with the increase of shikonin dose,the expression levels of NLRC4/Caspase-1/IL-1β/VEGFA were significantly down-regulated,confirming that shikonin affects psoriasis by regulating the expression of NLRC4.Conclusion: 1.The results of bioinformatics analysis confirmed that NLRC4 was highly expressed in psoriasis tissues in psoriasis patients and normal human skin tissues.2.In Ha Ca T keratinocytes stimulated by inflammatory factors TNF-α and IL-17 A,the expression of inflammasome NLRC4 was significantly increased.3.Gradient concentrations of shikonin can significantly reduce the expression of NLRC4/IL-1βpathway-related proteins in Ha Ca T keratinocytes stimulated by inflammation.The higher the concentration of shikonin,the more obvious the inhibition.4.Overexpression of NLRC4 can significantly promote the m RNA and protein expressions of Caspase-1,IL-1β and VEGFA.5.The therapeutic effect of shikonin on IMQ-induced psoriasis in mice is dose-dependent.With the increase in the dose of shikonin,the degree of skin lesions in IMQ-induced mice is reduced,and the excessive cell proliferation is weakened.Treatment is better.6.Different doses of shikonin can exert a therapeutic effect on the treatment of psoriasis by reducing the level of NLRC4/IL-1β pathway-related proteins.