目的:观察振心安神方(,ZXAS)对小鼠特应性皮炎(AD)的瞬时受体电位香草酸1(TRPV1)和瞬时受体电位锚蛋白1(TRPA1)信号通路的影响。
方法:用1-氯-2,4-二硝基苯(DNCB)对BALB/c小鼠的剃毛背侧皮肤诱导AD样病变。BALB/c小鼠分为5组:正常对照组,模型控制,西替利嗪,低,medium-,和高剂量的ZXAS。在ZXAS干预之后,收集皮损和血样进行苏木精和伊红染色,并测定炎性细胞因子的浓度.免疫球蛋白E(IgE),采用酶联免疫吸附试验(ELISA)检测白细胞介素(IL)-4,IL-5,IL-13和胸腺基质淋巴细胞生成素(TSLP)。收集脊髓用于测量胃泌素释放肽受体(GRPR)的表达,TRPV1和TRPA1通过免疫组织化学,西方印迹,和定量实时聚合酶链反应(qRT-PCR)分析。此外,3-(4,5-二甲基噻唑基-2)-2,5-二苯基四唑溴化物(MTT)测定,流式细胞术,ELISA,并进行蛋白质印迹分析原代背根神经节(DRG)神经元。
结果:ZXAS治疗通过降低皮炎评分改善DNCB诱导的AD样病变,划痕次数和表皮厚度,伴随着IgE和Th2炎性细胞因子的减少。ZXAS还抑制GRPR的mRNA和蛋白表达,脊髓中的TRPV1和TRPA1。ZXAS的含药血清减少辣椒素诱导的Ca内流,并下调DRG神经元中TRPV1,TRPA1和磷脂酶C的表达。
结论:ZXAS对AD的治疗作用可能与调节TRPV1、TRPA1和抑制神经元Ca2+信号有关。
To investigate the efficacy of Zhenxin Anshen formula (, ZXAS) on atopic dermatitis (AD) by transient receptor potential vanilloid 1 (TRPV1) and transient receptor potential ankyrin 1 (TRPA1) signalling pathway in mice and .
AD-like lesions were induced by 1-chloro-2,4-dinitrobenzene (DNCB) to the shaved dorsal skin of BALB/c mice. BALB/c mice were divided into five groups: normal control, model control, cetirizine, low-, medium-, and high-dose of ZXAS. After ZXAS in-tervention, the skin lesions and blood samples were collected for hematoxylin and eosin-stained and measuring the concentrations of inflammatory cytokines. Immun-oglobulin E (IgE), interleukin (IL)-4, IL-5, IL-13, and thymic stromal lymphopoietin (TSLP) were de-tected by Enzyme-linked immunosorbent assay (ELISA). The spinal cords were collected for measuring the expression of gastrin-releasing peptide receptor (GRPR), TRPV1, and TRPA1 by using immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction (qRT-PCR) analyses. In addition, 3-(4,5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, ELISA, and Western blotting were conducted for analysis of primary dorsal root ganglia (DRG) neurons .
ZXAS treatment improved DNCB-induced AD-like lesions through reducing dermatitis score, number of scratching and epidermal thickness, accompanied by the de-creased IgE and Th2 inflammatory cytokines. ZXAS also supressed the mRNA and protein expression of GRPR, TRPV1, and TRPA1 in the spinal cord. The medicated sera of ZXAS decreased capsaicin-induced Ca influx and downregulated the expression of TRPV1, TRPA1, and phospholipase C in DRG neurons.
The therapeutic effect of ZXAS on AD may be related to the regulation of TRPV1 and TRPA1 and inhibition of Ca2+ signals in neurons.