Effects of topical application of a tri-herb formula on inflammatory dry-skin condition in mice with oxazolone-induced atopic dermatitis,Phytomedicine

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Effects of topical application of a tri-herb formula on inflammatory dry-skin condition in mice with oxazolone-induced atopic dermatitis
Phytomedicine ( IF 6.7 ) Pub Date : 2021-07-30 , DOI: 10.1016/j.phymed.2021.153691
Xiao Xiao Wu ₁ , Wing Sum Siu ₁ , Chi Ling Wat ₁ , Chung Lap Chan ₁ , Chi Man Koon ₁ , Xiang Li ₁ , Wen Cheng ₁ , Hui Ma ₁ , Miranda Sin Man Tsang ₂ , Christopher Wai-Kei Lam ₃ , Ping Chung Leung ₁ , Clara Bik San Lau ₁ , Chun-Kwok Wong ₄

Affiliation

Background

Atopic dermatitis (AD) is a chronic relapsing inflammatory and pruritic skin disease, affecting 10-20% of the population worldwide. Paeonia suffruticosa Andrews (Paeoniaceae) (Cortex Moutan) and Mentha haplocalyx Briq. (Labiatae) (Herba Menthae) have shown beneficial effects on AD. Calendula officinalis L. (Asteraceae) is commonly used for treating skin rashes and wounds.

Purpose

In the present study, a three-herbs formula including Cortex Moutan and Herba Menthae, together with C. officinalis at 1:1:1 weight ratio was used as a topical agent and its therapeutic effects on AD was investigated.

Methods

In vitro effects of individual herbs and three-herbs formula (0.125–1 mg/ml) were examined using cytokine release assay on human mast HMC-1 cells, inflammation test on murine macrophage RAW cells and human keratinocyte (HaCaT) cells, and migration scratch assay on human umbilical vein endothelial cells (HUVEC). The contributing functional pathway of three-herbs formula in AD was explored using Western Blot assay in HMC-1 cells. Oxazolone-induced AD-like mice model was also used to investigate the in vivo therapeutic effect of the topical application of the three-herbs formula.

Results

Herba Menthae, Cortex Moutan, and three-herbs formula significantly reduced the production of IL-6 and tumor necrosis factor (TNF)-α in HMC-1 cells, inhibited the expression of IL-6, IL-8 and CCL2 in TNF-α/IFN-γ stimulated HaCaT cells, and suppressed the lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW 264.7 cells. Moreover, Herba Menthae and three-herbs formula significantly suppressed CCL2 and TNF-α production in LPS-induced RAW 264.7 cells. C. officinalis and three-herbs formula promoted wound healing in HUVEC. For intracellular mechanisms, three-herbs formula inhibited the expressions of molecules in STAT1 and STAT3-dependent pathways. In vivo model showed that topical application of three-herbs formula on challenged ear reduced ear swelling and mice scratching frequencies. H&E and toluidine blue staining of the challenged ear tissue demonstrated that three-herbs formula reduced the epidermal thickness and mast cell infiltration, respectively.

Conclusion

The three-herbs formula of Cortex Moutan, Herba Menthae and C. officinalis at 1:1:1 (w/w) exhibited anti-inflammatory effect and promotion of cell migration in vitro. It also alleviated ear redness, swelling, epidermal thickness and inflammation of the OXA-induced AD mice. These findings suggest a potential beneficial role of the topical application of the three-herbs formula for treatment of AD.

中文翻译：

外用三药方对恶唑酮致特应性皮炎小鼠炎症性干性皮肤病的影响

背景

特应性皮炎 (AD) 是一种慢性复发性炎症和瘙痒性皮肤病，影响全球 10-20% 的人口。Paeonia suffruticosa Andrews (Paeoniaceae) (Cortex Moutan) 和Mentha haplocalyx Briq。(唇形科) (Herba Menthae) 已显示出对 AD 的有益作用。Calendula officinalis L.（菊科）通常用于治疗皮疹和伤口。

目的

在本研究中，三草药配方包括牡丹皮和薄荷，连同C.巴戟以1：1：1个重量比被用作外用剂及其对AD的治疗效果进行了研究。

方法

使用对人肥大 HMC-1 细胞的细胞因子释放测定、对鼠巨噬细胞 RAW 细胞和人角质形成细胞 (HaCaT) 细胞的炎症测试以及迁移，检查了单个草药和三草药配方 (0.125–1 mg/ml) 的体外作用人脐静脉内皮细胞 (HUVEC) 的划痕试验。使用蛋白质印迹法在 HMC-1 细胞中探索了三草配方在 AD 中的作用功能途径。还使用恶唑酮诱导的AD样小鼠模型来研究局部应用三草方的体内治疗效果。

结果

薄荷、牡丹皮、三草配方显着降低HMC-1细胞中IL-6和肿瘤坏死因子(TNF)-α的产生，抑制TNF-α中IL-6、IL-8和CCL2的表达。 α/IFN-γ 刺激 HaCaT 细胞，并抑制 RAW 264.7 细胞中脂多糖 (LPS) 诱导的一氧化氮 (NO) 产生。此外，薄荷脑和三草本配方显着抑制 LPS 诱导的 RAW 264.7 细胞中 CCL2 和 TNF-α 的产生。厚朴和三草本配方促进 HUVEC 中的伤口愈合。在细胞内机制方面，三草方抑制STAT1和STAT3依赖途径中分子的表达。体内模型显示，在受攻击的耳朵上局部应用三草本配方可减少耳朵肿胀和老鼠抓挠的频率。受攻击的耳组织的 H&E 和甲苯胺蓝染色表明，三草本配方分别减少了表皮厚度和肥大细胞浸润。