清热祛湿方通过多靶点机制抑制特应性皮炎。

Qingre Qushi formula suppresses atopic dermatitis via a multi-target mechanism.

机构信息

Department of Dermatology, Shuguang Hospital Affiliated with Shanghai University of Traditional Chinese Medicine, Shanghai, China.

Shanghai Skin Disease Hospital, School of Medicine, Tongji University, Shanghai, China.

出版信息

J Ethnopharmacol. 2024 Jan 10;318(Pt A):116923. doi: 10.1016/j.jep.2023.116923. Epub 2023 Jul 22.

DOI:10.1016/j.jep.2023.116923

PMID:37487967

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Our previous studies have shown that the Qingre Qushi (QRQS) formula can treat atopic dermatitis (AD), and its possible mechanism is related to the regulation of the IL-33/ST2 signaling pathway. However, the molecular mechanism of AD is complex, and various AD subtypes respond better to therapies aimed at distinct targets.

AIM OF THE STUDY

This study aimed to investigate the multi-target mechanism of QRQS using experimental and network pharmacology studies.

MATERIALS AND METHODS

Flaky tail (FT) mice were treated with different concentrations of QRQS and cetirizine. The dermatitis score, scratching frequency, and histological evaluation were normatively evaluated. The levels of IgE and IgG1 in serum were tested using ELISAs. Using ELISA and RT-PCR, the expression of associated cytokines was determined. IL-17A-stimulated HaCaT cells were treated with QRQS to assess mRNA and protein expression. To elucidate the mechanism, a network pharmacology analysis based on active components derived from UPLC was conducted. Through molecular docking, we evaluated the binding affinity between the active constituents of QRQS and potential targets.

RESULTS

Using UPLC, 177 active ingredients in QRQS were identified. Network pharmacology analysis showed that the anti-AD effect of the active ingredients was related to the IL-17 signaling pathway and its related targets. FT mice are characterized by Th17-dominated immune disorders. QRQS ameliorated AD-like symptoms and decreased dermatitis scores and scratching frequencies. After QRQS treatment, IL-17A expression was inhibited and IL-17 pathway-associated cytokines were downregulated. Along with changes in Th17-differentiation, QRQS suppressed the expression of IL-4, IL-13, and TNF-α. QRQS also decreased the expression of IL-6, IL-8, and COX-2 in HaCaT cells exposed to IL-17A. The anti-AD active doses are 3.86 g/kg/day in vivo and 100 μg/mL in vitro.

CONCLUSION

QRQS has a multi-target immunoregulatory effect on AD and can improve the Th17-dominated inflammatory response by regulating the IL-17A signaling pathway. Quercetin, genistein, luteolin, and kaempferol are potential active ingredients.

摘要

民族药理学相关性

我们之前的研究表明，清热祛湿（QRQS）配方可以治疗特应性皮炎（AD），其可能的机制与调节 IL-33/ST2 信号通路有关。然而，AD 的发病机制复杂，各种 AD 亚型对针对不同靶点的治疗反应更好。

研究目的

本研究旨在通过实验和网络药理学研究探讨 QRQS 的多靶点机制。

材料与方法

用不同浓度的 QRQS 和西替利嗪处理特应性皮炎模型（FT）小鼠，对其皮肤炎症评分、抓挠频率和组织学评价进行规范评估。采用 ELISA 法检测血清 IgE 和 IgG1 水平。采用 ELISA 和 RT-PCR 法检测相关细胞因子的表达。用 QRQS 处理白细胞介素-17A（IL-17A）刺激的 HaCaT 细胞，检测 mRNA 和蛋白表达。基于 UPLC 得到的活性成分进行网络药理学分析，以阐明其作用机制。通过分子对接，评估 QRQS 活性成分与潜在靶点的结合亲和力。

结果

采用 UPLC 鉴定 QRQS 中的 177 种活性成分。网络药理学分析表明，活性成分的抗 AD 作用与 IL-17 信号通路及其相关靶点有关。FT 小鼠的特征是 Th17 为主的免疫紊乱。QRQS 改善 AD 样症状，降低皮炎评分和搔抓频率。经 QRQS 治疗后，IL-17A 表达受到抑制，IL-17 通路相关细胞因子下调。随着 Th17 分化的变化，QRQS 抑制了 IL-4、IL-13 和 TNF-α 的表达。QRQS 还降低了 IL-17A 作用下 HaCaT 细胞中 IL-6、IL-8 和 COX-2 的表达。体内抗 AD 活性剂量为 3.86 g/kg/天，体外为 100 μg/mL。