Jilin Ginseng Academy, Changchun University of Chinese Medicine, 1035 Boshuo Road, Changchun, Jilin Province 130117, China.
Jilin Ginseng Academy, Changchun University of Chinese Medicine, 1035 Boshuo Road, Changchun, Jilin Province 130117, China.
Phytomedicine. 2022 Oct;105:154372. doi: 10.1016/j.phymed.2022.154372. Epub 2022 Jul 31.
Huanglian ointment exhibits clinical efficacy for repairing skin barriers and inhibiting skin inflammation, and has been used to ameliorate eczema for many years. However, the active components and mechanism of Huanglian ointment have not yet been elucidated.
This study aimed to demonstrate the main active components and molecular mechanisms of Huanglian ointment for the treatment of eczema.
The main active components of Huanglian ointment were identified by gas chromatography-mass spectrometry. Network pharmacology approach and molecular docking techniques to predict the potential molecular mechanisms of Huanglian ointment alleviating eczema. Furthermore, Biostir-AD®-induced guinea pigs and tumor necrosis α (TNF-α)/interferon γ (IFN-γ)-induced HaCaT cells were employed to investigate the effectiveness and mechanisms of Huanglian ointment using histopathological staining, enzyme-linked immunosorbent assay, MTT assay, and western blot analysis.
Fourteen chemistry components were identified in Huanglian ointment. In total, 78 intersecting gene targets were identified between Huanglian ointment and eczema, including Jun, inflammatory regulators, and chemokine factors. Intersecting gene targets were enriched for cytokine and chemokine receptor binding, and inflammatory related signaling pathways. The molecular docking results showed that the identified components had a stable binding conformation with core targets. In vivo experiments showed that Huanglian ointment markedly ameliorated eczema-like skin lesions, restored histopathological morphology, and decreased levels of TNF-α, IFN-γ, and interleukin 6. Moreover, Huanglian ointment effectively protected HaCaT cells against TNF-α/IFN-γ-induced cell death and overproduction of thymus- and activation-regulated chemokine, macrophage-derived chemokine, and regulated upon activation normal T cell-expressed and secreted factor. Subsequently, we found that Huanglian ointment repaired skin barriers by affecting c-Jun, JunB, and filaggrin expression, and suppressed inflammatory response by inhibiting AGE-RAGE signaling pathway, both in vivo and in vitro.
Our results demonstrated that Huanglian ointment repaired skin barriers and inhibited inflammation by maintaining the balance of c-Jun and JunB, and suppressing AGE-RAGE signaling pathway, thereby relieving eczema. These findings providing a molecular basis for treatment of eczema by Huanglian ointment.
黄连软膏在修复皮肤屏障和抑制皮肤炎症方面具有临床疗效，多年来一直用于改善湿疹。然而，黄连软膏的活性成分和作用机制尚未阐明。
本研究旨在阐明黄连软膏治疗湿疹的主要活性成分和分子机制。
采用气相色谱-质谱联用技术鉴定黄连软膏的主要活性成分。采用网络药理学方法和分子对接技术预测黄连软膏缓解湿疹的潜在分子机制。此外，采用 Biostir-AD®诱导的豚鼠和肿瘤坏死因子-α（TNF-α）/干扰素-γ（IFN-γ）诱导的 HaCaT 细胞，通过组织病理学染色、酶联免疫吸附试验、MTT 分析和 Western blot 分析研究黄连软膏的疗效和作用机制。
黄连软膏中鉴定出 14 种化学成分。黄连软膏与湿疹的交集基因靶点共有 78 个，包括 Jun、炎症调节因子和趋化因子因子。交集基因靶点富集于细胞因子和趋化因子受体结合以及炎症相关信号通路。分子对接结果表明，鉴定出的成分与核心靶点具有稳定的结合构象。体内实验表明，黄连软膏能显著改善湿疹样皮肤损伤，恢复组织病理学形态，降低 TNF-α、IFN-γ和白细胞介素 6 的水平。此外，黄连软膏能有效保护 HaCaT 细胞免受 TNF-α/IFN-γ诱导的细胞死亡和胸腺激活调节趋化因子、巨噬细胞衍生趋化因子和激活正常 T 细胞表达和分泌因子的过度产生。随后，我们发现黄连软膏通过影响 c-Jun、JunB 和丝聚合蛋白的表达来修复皮肤屏障，并通过抑制 AGE-RAGE 信号通路来抑制炎症反应，无论是在体内还是体外。
我们的研究结果表明，黄连软膏通过维持 c-Jun 和 JunB 的平衡和抑制 AGE-RAGE 信号通路来修复皮肤屏障和抑制炎症，从而缓解湿疹。这些发现为黄连软膏治疗湿疹提供了分子基础。
