Mechanisms Of Different Effective Components Of Traditional Chinese Medicine In The Treatment Of Psoriasis From The Perspective Of Mesenchymal Stem Cells And Keratinocytes

BackgroundPsoriasis is a common clinical disease with recurrent attacks and lingering difficulties.Epidemiological studies have found that psoriasis affects about 2%of the world's population.The incidence of psoriasis in China is also very high,and the number of cases is increasing[1-2]The initial pathological changes of psoriasis are keratinocyte dysplasia,hyperkeratosis and incompleteness,accompanied by infiltration and aggregation of a variety of inflammatory-related cells.Because its pathogenesis is still unclear,there is no complete cure method at present.At present,it is generally believed that there is a close relationship between the occurrence of this disease and immune disorders.Most of them are abnormal levels of inflammatory cytokines in vivo and in the lesions,excessive activation of various immune cells.The interaction of the above several pathways eventually triggers an inflammatory waterfall,which enlarges the inflammatory reaction in the lesions step by step,leading to the occurrence of this disease.Stem cells(SC)are a kind of multipotential cells with self-differentiation and renewal ability,including embryonic stem cells(ESCs)and adult stem cells(ASCs).The former can be divided into hematopoietic stem cells(HSCs),mesenchymal stem cells(MSCs)and so on.Because MSCs have special immune regulation function and low immunogenicity,they are widely used in clinical treatment of many diseases related to autoimmune disorders.In addition,MSCs can also produce immune regulation on T lymphocyte by secreting many inflammatory-related cytokines,and help the body restore immune balance through its immune regulation.Keratinocyte(KC)is an immune-related cell,which plays an important role in the occurrence and development of psoriasis.The initial pathological changes of psoriasis are abnormal proliferation of KC with hyperkeratosis,proliferation of vascular endothelial cells and infiltration of many inflammatory-related cells.In addition,the occurrence of psoriasis is related to the uncontrolled apoptosis of keratinocytes,which shows that keratinocytes play a very important role in the occurrence and development of psoriasis.Traditional Chinese medicine has unique advantages and good clinical efficacy in the treatment of psoriasis.At present,the treatment of psoriasis mostly follows Zhu Renkang and Zhu Lao's treatment viewpoint,which classifies its syndrome types into"blood heat","blood stasis" and "blood dryness",and corresponds to the treatment methods of "cooling blood","promoting blood circulation" and "nourishing blood".At the same time,Dr.Song Ping inherited the academic ideas of many famous and old experts and combined with clinical experience,put forward another important pathogenesis of psoriasis,that is,"Xuanfu is depressed,kidney essence is deficient",and should be treated by "opening Xuanfu,invigorating kidney and peiyuan" method to achieve good clinical therapeutic effect.In addition,different medical schools still have different ideas and methods on understanding and treatment of psoriasis.To study the immune regulation of effective components of traditional Chinese medicine on MSCs,KC and its inflammatory response model,so as to combine traditional Chinese medicine with immunology,explore the possible mechanism of traditional Chinese medicine in the treatment of psoriasis,and combine mesenchymal stem cells,keratinocytes and the pathogenesis of psoriasis.Together,it enriches the connotation of traditional Chinese medicine treatment of psoriasis.Therefore,on the basis of inheriting the theory and clinical practice of "Kaixuan Bushen" therapy for psoriasis,combining with the understanding of the pathogenesis of psoriasis by contemporary physicians,summarizing and classifying the commonly used drugs,this topic proposes to explore the immune regulatory effects of different effective components of traditional Chinese medicine on MSCs,KC and their inflammatory response models,from MSCs,KC and psoriasis.Starting with the important cells related to psoriasis,we preliminarily studied the immune regulation effect of effective components of traditional Chinese medicine on psoriasis-related inflammatory response model from the perspective of integrated traditional Chinese and Western medicine,so as to preliminarily study the possible mechanism of effective components of traditional Chinese medicine in the treatment of psoriasis.In this study,cell proliferation,flow cytometry,RT-qPCR,ELISA and other techniques were used to study the effects of effective components of traditional Chinese medicine on cell proliferation and cell cycle of MSCs and KC,as well as the secretion of inflammatory cytokines and gene expression in inflammatory response model.The immune regulation of these effective components of traditional Chinese medicine on inflammatory response model of MSCs and KC was further studied.To explore the possible mechanism of different effective components of traditional Chinese medicine in the treatment of psoriasis.To sum up,based on previous data and Pre-team experimental studies,this project explores the immune regulation function of effective components of traditional Chinese medicine on inflammatory response models of mesenchymal stem cells,keratinocytes and mesenchymal stem cells,keratinocytes respectively,integrates clinical research and cytological research,and explores the possible immune mechanism of effective treatment of psoriasis by traditional Chinese medicine,enriches the treatment of psoriasis.Objective 1.To clarify the effects of tectoridin and aucubin on adipose mesenchymal stem cells(ADMSCs)from healthy people.Meenchymal stem cells(AMSCs)and their biological effects on inflammatory response models induced by LPS and Poly I:C,respectively.2.To explore the biological effects of astragaloside,Morinda officinalis polysaccharide and paeonol on KC and its inflammatory response model induced by IL-22,and to reveal the possible mechanism of effective components of traditional Chinese medicine in the treatment of psoriasis.Methods 1.AMSCs and KC were cultured in vitro and reserved for the third and twelfth generations respectively.The effective components of traditional Chinese medicine were selected to act on normal AMSCs and KC.Cell proliferation and cycle identification of AMSCs and KC were carried out by CCK-8 test and flow cytometry,and the corresponding drug concentration was screened out.2.RT-q PCR and ELISA were used to detect the expression of inflammatory factors and their mRNA in the inflammatory response model and establish the inflammatory response model.3.The selected effective components of traditional Chinese medicine were applied to the inflammatory response model.CCK-8 test,ELISA and RT-q PCR were used to investigate the effects of effective components of traditional Chinese medicine on the expression of inflammatory factors and their mRNAs in AMSCs and KC inflammatory response models,and to study the biological effects of effective components of traditional Chinese medicine on inflammatory response model.Result In the first part of the study,we found that the effective components of Chinese herbal medicine selected in this experiment could promote the proliferation of normal AMSCs except 25 ug/mL aucubin and 20 ug/mL atractydin(P<0,01 or P<0.001).atractylenolide 1,ginsenoside Rbl,tectordin and Oxymatrine could increase the G1 phase ratio of normal AMSCs and decrease the G2 or S phase ratio(P<0.05 or P<0.01);Paeonol and Astragaloside could reduce the healthy AMSCs.The proportions of G1 and S phase increased the proportions of G2 phase(P<0.01 or P<0.05).LPS 20ng/uL can increase the secretion level of IL-6 and IL-8 and the expression of IL-8 in the inflammatory response model,thus establishing the inflammatory response model of AMSCs.Poly I:C 25ug/mL can increase the expression level of IL-6,IL-8,TNF-a and TGF-? in the inflammatory response model of AMSCs,thus establishing the inflammatory response model.The effective components of Chinese herbal medicine,aucubin and aucubin,significantly promoted the proliferation of AMSCs in LPS-induced inflammatory response model(P<0.05 or P<0.001),and aucubin and tectordin promoted the proliferation of AMSCs in poly I:C-induced inflammatory response model(P<0.05).The levels of IL-8 and IL-1? in AMSCs inflammatory response model induced by LPS were significantly decreased by tectordin(50 ug/mL)and aucubin(100 ug/mL)(P<0.05 or P<0.01),and both of them could decrease the expression of IL-6 and IFN-y in AMSCs inflammatory response model induced by LPS(P<0.05 or P<0.01),and tectordin could decrease the expression of TNF-a(P<0.01),at the same time.Both of them have a tendency to increase the expression of TGF-?.tectoridin(50ug/mL)and aucubin(100 ug/mL)could decrease the level of IL-8 secretion in AMSCs inflammatory response model induced by poly I:C(P<0.001 or P<0.01).In the second part,we found that Morinda officinalis polysaccharide 25ug/mL,paeonol 10ug/mL,astragaloside 20ug/mL inhibited the proliferation of normal KC(P<0.001).IL-22 50ng/mL could increase the secretion level of IL-6 and IL-8 and the expression of IL-8 in KC inflammatory response model(P<0.01),so as to establish KC inflammatory response model.The above effective components of traditional Chinese medicine increased the expression of IL-6,IL-8,IL-17 and IL-1? in inflammatory response model.Conclusio 1.LPS induces healthy AMSCs to change to pro-inflammatory direction.The effective components of traditional Chinese medicine(TCM)such as aucubin and tectordin can reduce the secretion of IL-1?,IL-8,and the expression of TNF-a and IFN-y in inflammatory response model and alleviate inflammation.2.Poly I:C can induce the two-way transformation of healthy AMSCs to pro-inflammatory and anti-inflammatory.The effective components of Chinese herbal medicine,aucubin and Shegan glycoside,can reduce the level of IL-8 secretion in inflammatory response model and alleviate inflammation to some extent3.Astragaloside IV 20ug/mL,paeonol 10ug/mL,Morinda officinalis 25ug/mL Atracylenolide ?25ug/mL and Atracylenolide ?25ug/mL could inhibit the proliferation of keratinocytes.