振新安神方改善小鼠特应性皮炎样皮肤功能障碍及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 的调节。
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.
机构信息
Department of Dermatology, Shaanxi Provincial Hospital of traditional Chinese Medicine, Xi'an 710003, China.
Department of Dermatology, Xiyuan Hospital China Academy of Chinese Medical Sciences, Beijing 100193, China.
出版信息
J Tradit Chin Med. 2023 Oct;43(5):887-896. doi: 10.19852/j.cnki.jtcm.20230802.003.
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.
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，研究镇心安神方（ZXAS）对小鼠变应性皮肤病（AD）的疗效。
方法
用 1-氯-2,4-二硝基苯（DNCB）处理 shaved 背部皮肤诱导 AD 样病变。BALB/c 小鼠分为五组：正常对照组、模型对照组、西替利嗪、ZXAS 低、中、高剂量组。ZXAS 干预后，采集皮肤病变和血液样本，进行苏木精和伊红染色，并测量炎症细胞因子的浓度。酶联免疫吸附试验（ELISA）检测免疫球蛋白 E（IgE）、白细胞介素（IL）-4、IL-5、IL-13 和胸腺基质淋巴细胞生成素（TSLP）。免疫组化、Western blot 和实时定量聚合酶链反应（qRT-PCR）分析测量脊髓胃泌素释放肽受体（GRPR）、TRPV1 和 TRPA1 的表达。此外，还进行了 3-（4,5-二甲基噻唑基-2）-2,5-二苯基四氮唑溴盐（MTT）测定、流式细胞术、ELISA 和 Western blot 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、搔抓次数和表皮厚度改善 DNCB 诱导的 AD 样病变，同时降低 IgE 和 Th2 炎症细胞因子。ZXAS 还抑制脊髓中 GRPR、TRPV1 和 TRPA1 的 mRNA 和蛋白表达。ZXAS 的药物血清降低了辣椒素诱导的 Ca2+内流，并下调了 DRG 神经元中 TRPV1、TRPA1 和磷脂酶 C 的表达。
结论
ZXAS 治疗 AD 的疗效可能与调节 TRPV1 和 TRPA1 以及抑制神经元中的 Ca2+信号有关。
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