The Inflammation And Proliferation Effect About NE、 Trappin-2 On HaCat Cell And Intervention Of Chinese Herbal Compound—Psoriasis No.1

ObjectivePsoriasis is a common chronic inflammatory disease for dermatology, one of its main pathological characteristic is characterized by hyper proliferation of keratinocytes and polymorphonuclear neutrophils aggregation.Neutrophil elastase(NE) which is stored and commponded by PMNs is the focus of the current research.NE can cause tissue injury, inflammation, amplification, promoting proliferation, and it also play an important role in the pathogenesis of psoriasis. Trappin-2, an endogenous inhibitor of NE, is mainly secreted by keratinocytes and often expressed in some inflammatory skin diseases and skin tumors.It is the keratinocyte differentiation marker protein, through effective degradation can inhibit the activity of NE. At present, pathogenic theory about the imbalance of NE/trappin-2 has been widely recognized.Human immortalized cutin cells (HaCat cells) is extremely similar to the basic characteristics of KC, has been widely used in the study of psoriasis. In this paper, we applied the TNF-a-stimulated HaCat cell which has similar pathological features of psoriasis by different ce-ntrations of traditional Chinese medicine compound——Psoriasis NO.1, and we observe the secretion of NE, trappin-2 and related cytokines (IL-6, IL-8 and ICAM-1, etc) expression level, In order to explore mechanisms of NE, trappin-2 in the pathogenesis of psoriasis,to clarify Chinese Medicine compound effect on psoriasis intrinsic material base and provide favorable scientific basis for new target to Traditional Chinese Medicine compound in treatment of psoriasis.MethodsThe experiment set up different concentrations of traditional Chinese medicine compound group, tripterygium wilfordii group and normal saline group, and each group containing medicine serum preparation. HaCat cells were cultured in vitro and inoculated in the 96-well plates, when cells covered the bottom of the hole bottom, each group containing serum on the culture plates. In addition to the normal saline group, other groups were added to the same amount of TNF-α. Respectively, cultured at different points of time.Using ELISA method to detect NE, trappin-2, IL-6, IL-8 levels and Western blot method to detect the expression of ICAM-1 level.Results1、NE experimental results displayed:Psoriasis NO.1(concentrate) group and Psoriasis NO.1(1/2) concentration group on 12h was statistically significant (P<0.05), and the highest concentration of NE and 1/2 concentration group was the lowest. All time points of Psoriasis NO.1(1/2) concentration group, the results at 12h get the lowest concentration, and compared with that at 0h,24h,48h, it was statistically significant (P<0.05);2、trappin-2 experimental results showed:Psoriasis NO.1 (1/2) group、 Psoriasis NO.1(1/4) group and other groups was statistically significant (P< 0.05), that at 0h,12h,24h,48h between the two groups was statistically significant in Psoriasis NO.1(1/4) group (P<0.05); Oh trappin-2 was the lowest, the highest concentration of trappin-2 is 48h;3、IL-6 experimental results showed:Psoriasis NO.1(1/4) group and other groups was statistically significant (P<0.05). that at 0h,12h, 48h was statistically significant (P< 0.05) respectively in the Psoriasis NO.1(1/4) group;4、IL-8 experimental results showed:Psoriasis NO.1(1/4) group and other groups was statistically significant (P<0.05). that at 0h,12h, 24h was statistically significant (P< 0.05) respectively in the Psoriasis NO.1(1/4) group;5、Western blot method results displayed between the groups were not statistically significant(P>0.05). Conclusion1、Traditional Chinese Medicine on NE, trappin-2 did not change with the change of the concentration.The effect of Psoriasis NO.1(1/2) group inhibited the expression of NE was the strongest at 12h and the action was induced over time. Psoriasis NO.1 (1/4) group inhibited the expression of trappin-2 was the strongest at 24h and the action was induced over time.2、TCM compound for IL-6, IL-8 was no correlation between the concentration. Psoriasis NO.1(1/4) group inhibited the expression of IL-6, IL-8 was the strongest at 24h and the action was induced over time.In summary, Traditional Chinese Medicine compound may, through inhibiting NE、trappin-2、IL-6 and IL-8 expression level, reduce the inflammatory response, clarify the role of NE and trappin-2 in the pathogenesis of psoriasis and provide new targets for traditional Chinese medicine compound to the treatment of psoriasis mechanisms.