开玄解毒核心方通过调控神经-免疫轴抑制Th17细胞皮肤归巢改善IMQ联合束缚应激诱导的小鼠银屑病的机制OA

Objective:To observe the effect and therapeutic effect of Kaixuan Jiedu core prescription(KXJD)on skin homing of Th17 cells in the mouse model of imiquimod(IMQ)combined with restraint stress-induced psoriasis-like skin damage,and to explore its potential mechanism from the perspective of neuro-immune axis.Methods:Thirty male C57BL/6J mice were randomly allocated into five groups(n=6):Control,model(IMQ),restraint stress model(IMQ+RS),KXJD,and methotrexate(MTX).The mouse model of psoriasis-like skin damage was established by 5%IMQ combined with restraint stress.At the same time of modeling,each treatment group was treated with corresponding doses of drugs,and the control,IMQ,and IMQ+RS groups were treated with the same amount of normal saline by gavage once a day for 5 days.Hematoxylin-eosin(HE)staining was used to observe the pathological changes in the skin tissue and Baker scoring was performed.Serum levels of interleukin-1β(IL-1β)and angiopoietin-2(Ang-2)were measured by enzyme-linked immunosorbent assay(ELISA).The levels of matrix metalloproteinase-9(MMP-9),tissue inhibitor of metalloproteinase-1(TIMP-1),C-C motif chemokine ligand 20(CCL20),and C-C motif chemokine receptor 6(CCR6)in the skin tissue were determined.Immunohistochemistry(IHC)was employed to determine the protein expression of cutaneous lymphocyte-associated antigen(CLA),integrin αE(CD103),cytokeratin 10(CK10),and nuclear factor-kappa B(NF-κB)in the skin.Immunofluorescence double staining(DIF)was adopted to detect the expression and co-localization of vascular endothelial cadherin(VE-cadherin)and platelet-endothelial cell adhesion molecule(CD31),CCR6,CD103,substance P(SP),calcitonin gene-related peptide(CGRP),and protein gene product 9.5(PGP9.5)in the skin tissue.Real-time PCR was employed to quantify the mRNA levels of IL-10,IL-17A,and IL-23.Results:Compared with the control group,the IMQ group and IMQ+RS group showed significant inflammatory cell infiltration,abnormal proliferation of epidermal cells,keratinization and other pathological changes in the skin tissue,and a significant increase in Baker score,elevated levels of IL-1β and Ang-2 in the serum and MMP-9,CCL20 and CCR6 in the skin lesions,upregulated expression of CLA,CD103,CK10,NF-κB,IL-17A mRNA,and IL-23 mRNA in the skin lesions,and downregulated expression of TIMP-1 and IL-10 mRNA.In addition,the fluorescence intensities of VE-cadherin and CD31 co-localization,CCR6 and CD103 co-localization,SP and PGP9.5 co-localization,and CGRP and PGP9.5 co-localization were increased(P<0.05).Compared with the IMQ group,the above indicators in the IMQ+RS group were further aggravated.Compared with the IMQ group,the above indicators in the IMQ+RS group were further aggravated.Compared with the IMQ+RS group,KXJD and MTX significantly alleviated the pathological damage of skin lesions,significantly decreased the Baker score,lowered the levels of IL-1β and Ang-2 in the serum and MMP-9,CCL20 and CCR6 in skin lesions,downregulated the expression of CLA,CD103,CK10,NF-κB,IL-17A mRNA and IL-23 mRNA in skin lesions,and upregulated the expression of TIMP-1 and IL-10 mRNA.Furthermore,KXJD and MTX reduced the fluorescence intensities of VE-cadherin and CD31 co-localization,CCR6 and CD103 co-localization,CGRP and PGP9.5 co-localization,and SP and PGP9.5 co-localization(P<0.05).Conclusion:KXJD can significantly ameliorate the psoriasis-like skin damage induced by IMQ combined with restraint stress in mice by regulating the neural-immune axis and inhibiting the skin homing of Th17 cells.