Putuo Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200062, China; Shanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Shanghai Frontiers Science Center of TCM Chemical Biology, Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai, 201203, China.
Phytomedicine. 2022 Aug;103:154228. doi: 10.1016/j.phymed.2022.154228. Epub 2022 Jun 2.
Atopic dermatitis (AD), a common inflammatory skin disorder, severely affects the life quality of patients and renders heavy financial burden on patient's family. The Chinese medicine Viola yedoensis Makino formula (VYAC) has been widely used for treating various skin disorders. Previous studies have reported that VYAC is effective in relieving DNCB-induced AD and inflammation. However, the anti-inflammatory mechanism of VYAC is still ill-defined and poorly understood. This study aims to investigate the therapeutic effects of VYAC on DNCB-induced AD and to elucidate the underlying anti-inflammatory mechanisms.
VYAC were extracted with 70% ethanol and lyophilized for use. AD mice were established by DNCB. The therapeutic effects of VYAC were evaluated by oral administration VYAC (150, 300 and 600 mg/kg) daily in vivo. The histopathological and immunohistochemistry were used to analyze skin lesion and macrophages infiltration, RT-qPCR and Elisa were used to analyze the inflammatory factors in skin tissues and serum. To explore the underlying mechanism of VYAC against AD in vitro. RAW264.7 cells and bone-marrow-derived macrophages (BMDMs) were employed for macrophage polarization analysis. Flow cytometer, immunofluorescence and western blot were used to analyze M2 macrophages markers. STAT3 siRNA were transfected into both cells to validate the effects of VYAC-induced macrophages M2 polarization via JAK2/STAT3 signaling pathway.
VYAC ameliorated skin lesion of DNCB-induced AD mice by decreased clinical scores and epidermal thickness, decreased the level of pro-inflammatory factors (IL-1β, TNF-α and IL-18) and enhanced IL-10 anti-inflammatory factor level, inhibited macrophages infiltration and promoted M2 macrophages polarization in vivo. VYAC significantly promoted M2 macrophages polarization in vitro. It is observed that VYAC not only inhibited the phosphorylation of JAK2 and STAT3 in RAW264.7 cells and BMDMs, but also accelerated the translocation to the nucleus. What's more, VYAC reduced the polarization of M2 macrophage by activating JAK2/STAT3 signaling pathway was observed in both cells.
Our findings demonstrate that VYAC significantly ameliorates skin lesion of DNCB-induced AD mice and reduces the levels of inflammatory factors by activating JAK2/STAT3 signaling pathway and promoting M2 macrophages polarization.
特应性皮炎（AD）是一种常见的炎症性皮肤疾病，严重影响患者的生活质量，并给患者家庭带来沉重的经济负担。中药 Viola yedoensis Makino 配方（VYAC）已广泛用于治疗各种皮肤疾病。先前的研究报告称，VYAC 可有效缓解 DNCB 诱导的 AD 和炎症。然而，VYAC 的抗炎机制仍不清楚。本研究旨在探讨 VYAC 对 DNCB 诱导的 AD 的治疗作用，并阐明其潜在的抗炎机制。
采用 70%乙醇提取 VYAC 并冻干备用。DNCB 诱导 AD 小鼠模型。体内通过口服 VYAC（150、300 和 600mg/kg）每日给药评价 VYAC 的治疗效果。采用组织病理学和免疫组织化学分析皮肤病变和巨噬细胞浸润，采用 RT-qPCR 和 Elisa 分析皮肤组织和血清中的炎症因子。为了探讨 VYAC 体外抗 AD 的作用机制，采用 RAW264.7 细胞和骨髓来源的巨噬细胞（BMDMs）进行巨噬细胞极化分析。流式细胞术、免疫荧光和 Western blot 用于分析 M2 巨噬细胞标志物。STAT3 siRNA 转染到两种细胞中，通过 JAK2/STAT3 信号通路验证 VYAC 诱导的巨噬细胞 M2 极化的作用。
VYAC 通过降低临床评分和表皮厚度改善 DNCB 诱导的 AD 小鼠的皮肤病变，降低促炎因子（IL-1β、TNF-α 和 IL-18）水平，增加抗炎因子 IL-10 水平，抑制巨噬细胞浸润，促进体内 M2 巨噬细胞极化。VYAC 显著促进体外 M2 巨噬细胞极化。观察到 VYAC 不仅抑制 RAW264.7 细胞和 BMDMs 中 JAK2 和 STAT3 的磷酸化，而且还加速向核内转移。更重要的是，VYAC 通过激活 JAK2/STAT3 信号通路减少了两种细胞中 M2 巨噬细胞的极化。
我们的研究结果表明，VYAC 通过激活 JAK2/STAT3 信号通路和促进 M2 巨噬细胞极化，显著改善 DNCB 诱导的 AD 小鼠的皮肤病变并降低炎症因子水平。
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