Zhenxin Anshen formula ameliorates atopic der-matitis-like skin dysfunction in mice and regulation of transient receptor potential vanilloid 1 and transient receptor potential ankyrin 1 in Neural pathways.

Zhao Yiding, Yan Xiaoning, Jiang Shanshan, Liu Yong, Dong Chun, Chi Huiyan, Mao Chaoyi
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Abstract

Objective: 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 .

Methods: 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 .

Results: 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.

Conclusions: The therapeutic effect of ZXAS on AD may be related to the regulation of TRPV1 and TRPA1 and inhibition of Ca2+ signals in neurons.

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镇心安神方可改善小鼠特应性皮炎样皮肤功能障碍,调节神经通路中瞬时受体电位香草素1和瞬时受体电位锚蛋白1。
目的:通过瞬时受体电位香草醛1(TRPV1)和瞬时受体电位锚蛋白1(TRPA1)信号通路研究珍心安神方对小鼠特应性皮炎(AD)的治疗作用。将BALB/c小鼠分为5组:正常对照组、模型对照组、西替利嗪组、低剂量、中剂量和高剂量ZXAS组。ZXAS干预后,收集皮肤病变和血液样本进行苏木精和伊红染色,并测量炎性细胞因子的浓度。采用酶联免疫吸附试验(ELISA)检测免疫球蛋白E(IgE)、白细胞介素-4、IL-5、IL-13和胸腺基质淋巴细胞生成素(TSLP)。收集脊髓,通过免疫组织化学、蛋白质印迹和定量实时聚合酶链式反应(qRT-PCR)分析来测量胃泌素释放肽受体(GRPR)、TRPV1和TRPA1的表达。此外,还对原代背根神经节(DRG)神经元进行了3-(4,5-二甲基噻唑基-2)-2,5-二苯基四唑溴化物(MTT)法、流式细胞术、ELISA和Western印迹分析,伴随着IgE和Th2炎性细胞因子的降低。ZXAS还抑制脊髓中GRPR、TRPV1和TRPA1的mRNA和蛋白表达。ZXAS的药物血清减少了辣椒素诱导的钙内流,并下调了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和TRPA1以及抑制神经元内Ca2+信号有关。
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