The Effect Of No.1 Yinxie Formula To Secrete TNF-α And IL-8 In Psoriasis Serum Neutrophils Cultivated In Vitro Culture

ObjectivePsoriasis is an unexplained chronic,inflammatory skin disease,including psoriasis arthritis,pustular psoriasis,erythroderma psoriaticum,and the most common type,psoriasis vulgaris,accounting for more than 90%.It has the characteristics of easy recurrence,difficult to cure,and the main pathological features of keratinocyte hyperplasia,epidermal neutrophilic infiltration and neovascularization.The incidence of this disease is about0.1% ～ 2%,while in some European countries up to 3%.The disease caused greater physical and psychological burden on patients and their families.At present,the cause of the disease is not clear.Many studies have shown that the occurrence and development of the disease are related to many factors such as inheritance,immunity,infection,endocrine,neuropsychiatric,living habits,medicine and environment.For many years,many dermatologists regard psoriasis as a key research area in dermatology.Numerous studies have shown that both neutrophils and inflammatory cytokines are involved in the pathogenesis of psoriasis.Neutrophils are one of the main effector cells of nonspecific inflammatory reaction.The accumulation of neutrophils in the lesion is closely related to the active mitosis of epidermal cells,indicating that neutrophils play an important role in the development and progression of psoriasis.In recent years,it was found that neutrophils not only can produce a variety of immunomodulatory proteases,but also to secrete cytokines,such as TNF-α、IL-8、IFN-γ and IL-4.The pathogenesis of how neutrophil involvement in psoriasis it is not clear.The No.1 Yinxie Formula developed by our school has obtained good curative effect in the early animal experiment and clinical observation,and has the value of further clinical promotion.In this study,in order to further explore the mechanism of No.1 Yinxie Formula-treatment of psoriasis vulgaris syndrome,we studied the effects of different concentrations of serum on the secretion of TNF-α and IL-8 by neutrophils secreted in vitro.We explored its intrinsic material basis and anti-inflammatory mechanism in the treatment of psoriasis,in order to establish a new therapeutic target for traditional Chinese medicine compound to psoriasis,so as to provide new ideas of the treatment of psoriasis.MethodsFrist,we collected 18 patients cases of psoriasis vulgaris patients,which had been diagnosed clinically and pathologically.Their peripheral blood is drawn and the neutrophils are isolated and cultured using a lymphocyte separation fluid density gradient.The activity and purity of the cells were identified by trypan blue exclusion assay,and neutrophils were seeded on the culture plate.Second,we added the experimental groups of four concentrations of No.1Yinxie Formula,the tripterygium glycosides group and the normal saline group to the neutrophil culture medium,all above we added were serum containing drugs.After being cultured for 48 hours,we detected the expression level of secreted TNF-α and IL-8 in vitro cultured neutrophils from each group by ELISA method.ResultsThe expression of TNF-α: Compared with the normal saline group,the Chinese medicine liquid group,1/2 and 1/8 concentration group,as well as the tripterygium glycosides group were all statistically significant(P<0.05).Compared with tripterygium glycosides group,1/2,1/4 and 1/8 concentration group were all statistically significant(P<0.05).The expression of IL-8: The level of IL-8 in tripterygium glycosides group was significantly lower than other 5 groups,which was statistically significant(P <0.05),while the expression of IL-8 in normal saline group,1/2 concentration group and 1/8 concentration group were approximate,which were not statistically significant(P> 0.05).The expression of IL-8 between the Chinese medicine liquid group and the 1/4 concentration group were similar,with no significant difference(P> 0.05),but higher than the other groups,which was statistically significant(P <0.05).ConclusionIn this experiment,we found that No.1 Yinxie Formula can reduce the expression of TNF-α,while the level of IL-8 expression was not significantly reduced.It is speculated that the No.1 Yinxie Formula has a good curative effect in the treatment of psoriasis vulgaris.The mechanism may be that No.1Yinxie Formula can down-regulate the expression of TNF-α in peripheral blood neutrophils,thereby improving the psoriasis inflammatory response.