Inhibitory Effects And Preliminary Mechanisms Of Natural Plant Antimicrobial Solution(PAMs)on Inflammatory Skin Disease

The skin diseases seriously affect health,especially skin inflammatory disease and tumor.Patients are often difficult to be cured by traditional methods,and which can damage the body because of the side effects.However,Chinese traditional medicines have little side effects and improve the quality of life.Therefore,Chinese traditional medicines are more and more appreciated.Natural plant antimicrobial solution(PAMs)was extracted from folk medicinal plants in China and has many kinds of efficacy.PAMs have been widely used in folk and also has good development prospect for the research.By using modern molecular biology,we provide a molecular identification technique for the medicinal materials in PAMs,and the quality safety can be guaranteed from the sources of medicinal materials.At last,we proposed for PAMs's anti-inflammation effects of molecular mechanisms and further explain gene regulation of skin melanoma.The drug safety and molecular mechanism of PAMs are more improved.Firstly,we extract genomic DNA of Carthamus tinctorius?Arnebia euchroma?Blumea balsamifera and Solanum indicum.Then Analyze the differences between the easily-confused species' ITS2 sequences that are amplified through PCR,which procedure can help us study the identifying ability of ITS2 sequences.The result shows that,there is a C-T mutant at the location of 57th base of Carthamus tinctorius ITS2 sequence.Furthermore,we find C-T and T-C mutants at the location of 213 and 227 in the Blumea balsamifera ITS2 sequence respectively.Oppositely,there is no mutant of base has found in Solanum indicum and Arnebia euchroma ITS2 sequences.The difference analysis of ITS2 sequences of the four medical materials and their proximal species reveals that,the base differences between the ITS2 sequences of proximal species are much bigger than those of interspecific ITS2 sequences.The evolutionary tree shows that the four medical materials in the context have been divided into a class,which indicates the high-particularity of ITS2 and ITS2 sequences can be regarded as a category of DNA barcode sequences.Secondly,we confirmed the anti-inflammatory functions and proposed the mechanisms in vivo and vitro.The cytokines and proteins including IL-6,IL-8,IL-23,TNF-? and ICAM-1 were all suppressed by PAMs in TNF-?/IFN-?-induced HaCaT cells and imiquimod-induced psoriasis-like mice.PAMs ameliorates the psoriasis-like symptoms by imiquimod-induced psoriasis-like mice.PAMs also significantly ameliorate the psoriatic-like symptoms in mouse model,the evaluation scores for both the single(scales,thickness,erythema)and cumulative features were in the order of blank control<Dexamethasone<PAMs<50%ethanol<model groups,the latter was further confirmed by H&E staining,qRT-PCR and immunohistochemistry.PAMs suppressed the gene expression of IL-8,TNF-a,ICAM-1 and IL-23 in mouse tissues was consistent with the results from those of the HaCaT cells.Particularly,the inhibition of psoriasis-like skin inflammation by PAMs was correlated with the inactivation of P65 protein translocation in cellular nucleus indicating the inhibition of the inflammatory NF-?B signal pathway.All these findings suggested that PAMs may be a promising drug candidate for the treatment of inflammatory skin disorders,such as psoriasis.Then,we treated B16 cells with PAMs for 24 h,the differentially genes expression of blank group and drug group was analyzed by transcriptome sequencing.The statistical results show that 919 genes are up-regulation,1412 genes are down-regulation and part of the genes is validated through qRT-PCR.The mechanism of action of PAMs for melanoma was revealed from gene level.