Study On The Regulatory Effect Of Traditional Chinese Medicine For Nourishing Blood And Detoxification On The Tight Junctions Between Epidermal Cells In Psoriasis

Psoriasis is an autoimmune disease characterized by erythema,squamous and in-filtrating skin.The pathogenesis of psoriasis is closely related to the impairment of skin barrier function.Abnormal tight junction between keratinocytes can lead to its abnormal proliferation and differentiation,affect epidermis normal barrier function and promote the migration and infiltration of inflammatory cells and the secretion of inflammatory factors,which attack and destroy the tight junction,forming a vicious circle and maintaining the chronic inflammatory response at the skin lesions.In TCM Psoriasis blood dryness syndrome is mainly caused by Yin blood consumption for the wind,hot and dryness of suffering a long time,skin can not nourished.herb medi-cine nourishing blood anddetoxification prescription is its effective prescription.In this study,it was aimed to observe the regulatory effects of YangXue JieDu decoction,its Compatibility combination and the main component ferulic acid on the epidermal permeability barrier and the tight junction between epidermal cells in vivo and in vitro experiments.The aim is to elucidate the mechanism of YangXue JieDu prescription and its main components on the effect of tight juction between keratino-cytes and epidermal barrier in psorisasis,so as to provide a biological basis for herbal medicine clinical application.EXPERIMENT?:Regulation of YangxueJiedu Decoction to Tight Junction of Keratinocytes in PsoriasisObjective:To observe the expression of tight junction proteins(Claudin-1?Claudin-7?Occludin)of psoriasis-like lesions in mice,and to clarify the effect of Yangxue Jiedu decoction on the epidermal barrier of psoriasis.Methods:C57BL/6J mice were randomly divided into Control group?Model group?positive drug(methotrexate)group and Yangxue Jiedu decoction group.The mice were given imiquimot to induce psoriasis-skin lesions after the hair was shaved.Daily photos were taken to record the forms of skin lesions and PASI scores.Water and oil test pen was used to detect skin moisture content;The pathological changes were ob-served by htoxylin eosin(HE)staining and the epidermal thickness was measured.Ki67 was detected by immunofluorescence.Immunohistochemistry was used to detect the expression of Loricrin,CD3+T lymphocyte infiltration and Claudin-1?Claudin-7?Occludin.In addition to this,the expressions of Claudin-7 and Occludin in skin le-sions were detected by Western Blot.Results:Yangxue Jiedu decoction could significantly reduce the psoriasis skin lesions in mice,reduce the PASI score and skin thickness(P<0.01).Compared with the Model group,the skin moisture content in the lesion was increased(P<0.01).Abnormal ex-pression of Ki67 and Loricrin in epidermis and infiltration of CD3+T cells were re-duced(P<0.01).In addition,it increased the expression of Claudin-I.Claudin-7?Occludin proteins(P<0.05),and increased the integrity of the tight junction structure.Conclusion:By regulating the expression of tight junction proteins between ker-atinocytes,Yangxue Jiedu decoction can inhibit the abnormal proliferation?diff-erentiation and further restore the broken epidermal barrier.EXPERIMENT?:Regulation of YangxueJiedu Decoction to Tight Junction in HaCaT cells.Objective:To observe the effect of Yangxue jiedu decoction and its component on the expression of Claudin-1?Claudin-7 and Occludin on HaCaT cells.Methods:IL-17(1?g/ml)was uesd to simulate the microenvironment of psoriasis skin lesions.The intervention was conducted by making Yangxue component,Jiedu component and Yangxue Jiedu dry powder.The toxicity of the drug to HaCaT cells was detected by cell counting kit-8(CCK-8)method.The effect of drugs on the ex-pression of Claudin-1?Claudin-7?Occludin in HaCaT was detected by immunoflu-orescence assay.Results:In vitro studies,compared with the Model group,the expressions of Clauin-1?Claudin-7?and Occludin in the Yangxue Jiedu group and Yangxue group were in-creased(P<0.05).Compared with the Model group,there was no statistically signifi-cant difference in protein expression in the Jiedu group.Conclusion:The YangxueJiedu decoction plays a role in regulating tight junction mainly through the Yangxue component.EXPERIMENT ?:Regulatory effects of ferulic acid on skin lesions and tight junction between epidermal cells in psoriasis-like mice.Objectives:On the basis of previous experiments,the intervention effect of Yangxue component on the tight junction between epidermal cells was further explored.In this study,Ferulic Acid(FA),one of the main components of angelica sinensis,was se-lected to further explore its intervention effect on the tight junction between epidermalcells in psoriasis-like mice.Methods:BALB/c mice were randomly divided into Control group?Model gro-up?positive drug(methotrexate)group and high dose group of ferulic acid(F A-100 mg/kg),ferulic acid dosage group(FA-50 mg/kg),low dose group of ferulic acid(FA-25 mg/kg).The mice were given imiquimot to induce psor-iasis-skin lesions after the hair was shaved.Daily photos were taken to record the forms of skin lesions and PASI scores.Water and oil test pen was used to detect skin moisture content.The pathological changes were observed by htoxyli-n eosin(HE)staining and the epidermal thickness was measured.Ki67?loricrin?Claudin-1 and Claudin-7 was detected by immunofluorescence.Immunohistochem-istry was used to detect the expression of CD3+T lymphocyte infiltration andOccludin.Results:The high?medium and low doses of ferulic acid significantly reduced the psoriasis skin lesions in mice,and reduced the PASI score and skin thickness(P<0.01).Compared with the Model group,the skin moisture content in the high and me-dium dose of ferulic acid groups was increased(P<0.01).Abnormal expression of Ki67 and Loricrin in epidermis were reduced(P<0.01).In addition,ferulic acid increased the expression of Claudin-1?Claudin-7?Occludin proteins(P<0.05),and increased the integrity of the tight junction structure.The infiltration of CD3+T cells in the dermiswas reduced in the high and medium dose ferulic acid group(P<0.01).Conclusion:By regulating the expression of tight junction proteins between keratino-cytes,ferulic acid can inhibit the abnormal proliferation and differentiation and fur-ther restore the broken epidermal barrier.EXPERIMENT ?:The effect of ferulic acid on the tight junction between Ha-CaT cellsObjective:To observe the intervention of ferulic acid on the formation of tight junc-tion between HaCaT cells and to explore its possible mechanism.Methods:IL-17(1?g/ml)was used to simulate the microenvironment of psoriasis skin lesions.High?medium and low concentrations of ferulic acid(100?g/ml,50?g/ml,25?g/ml)were added for intervention.Cell proliferation toxicity assay kit-8(CCK-8)was used to detect the toxicity of HaCaT cells.Transwell plate was used to detect the permeability of HaCaT cells to fluorescent yellow.The effect of drugs on the expres-sion of Claudin-1?Claudin-7?Occludin in HaCaT was detected by immunofluores-cence assay.The expression levels of Cdc42 and p-aPKC in HaCaT cells were detect-ed by Western blot.Results:IL-17 can cause damage to the integrity of tight junction between HaCaT cells,and increase the permeability of HaCaT cells to fluorescent yellow.IL-17 also decreases the expressions of Claudin-1?Claudin-7 and Occludin.The expression levels of Cdc42 and p-aPKC were decreased.Compared with the Model group,the permea-bility of ferulic acid in the high-dose group(FA-100?g/ml)significantly decreased(P<0.05).The expression levels of Claudin-1?Claudin-7 and Occludin in the high-dose and medium-dose ferulic acid groups(FA-100?g/ml and FA-50?g/ml)could be in-creased to varying degrees,and the integrity of the integrity could be increased.The expression level of Cdc42 in the high-dose group(FA-100?g/ml)was significantly higher than that in the Model group(P<0.01),and the phosphorylation level of aPKCwas also significantly higher than that in the Model group(P<0.01).Conclusion:Ferulic acid can increase the expression level of Cdc42 and phosphory-lation level of aPKC,promoting the expression of tight junction protein.Ferulic acid can antagonize the damage of IL-17 to the tight junction between HaCaT cells,so as to achieve the purpose of treating psoriasis.