Natural Product Research Center, Korea Institute of Science and Technology (KIST), Gangneung 25451; Division of Bio-Medical Science & Technology, University of Science and Technology (UST), Daejeon 34113, Korea.
Division of Bio-Medical Science & Technology, University of Science and Technology (UST), Daejeon 34113; Natural Product Informatics Research Center, Korea Institute of Science and Technology (KIST), Gangneung 25451, Korea.
BMB Rep. 2022 Jun;55(6):275-280. doi: 10.5483/BMBRep.2022.55.6.144.
The treatment of atopic dermatitis (AD) is challenging due to its complex etiology. From epidermal disruption to chronic inflammation, various cells and inflammatory pathways contribute to the progression of AD. As with immunosuppressants, general inhibition of inflammatory pathways can be effective, but this approach is not suitable for long-term treatment due to its side effects. This study aimed to identify a plant extract (PE) with anti-inflammatory effects on multiple cell types involved in AD development and provide relevant mechanistic evidence. Degranulation was measured in RBL-2H3 cells to screen 30 PEs native to South Korea. To investigate the anti-inflammatory effects of Parasenecio auriculatus var. matsumurana Nakai extract (PAE) in AD, production of cytokines and nitric oxide, activation status of FcεRI and TLR4 signaling, cell-cell junction, and cell viability were evaluated using qRT-PCR, western blotting, confocal microscopy, Griess system, and an MTT assay in RBL-2H3, HEK293, RAW264.7, and HaCaT cells. For in vivo experiments, a DNCBinduced AD mouse model was constructed, and hematoxylin and eosin, periodic acid-Schiff, toluidine blue, and F4/80-staining were performed. The chemical constituents of PAE were analyzed by HPLC-MS. By measuring the anti-degranulation effects of 30 PEs in RBL-2H3 cells, we found that Paeonia lactiflora Pall., PA, and Rehmannia glutinosa (Gaertn.) Libosch. ex Steud. show an inhibitory activity of more than 50%. Of these, PAE most dramatically and consistently suppressed cytokine expression, including IL-4, IL-9, IL-13, and TNF-α. PAE potently inhibited FcεRI signaling, which mechanistically supports its basophil-stabilizing effects, and PAE downregulated cytokines and NO production in macrophages via perturbation of toll-like receptor signaling. Moreover, PAE suppressed cytokine production in keratinocytes and upregulated the expression of tight junction molecules ZO-1 and occludin. In a DNCB-induced AD mouse model, the topical application of PAE significantly improved atopic index scores, immune cell infiltration, cytokine expression, abnormal activation of signaling molecules in FcεRI and TLR signaling, and damaged skin structure compared with dexamethasone. The anti-inflammatory effect of PAE was mainly due to integerrimine. Our findings suggest that PAE could potently inhibit multi-inflammatory cells involved in AD development, synergistically block the propagation of inflammatory responses, and thus alleviate AD symptoms. [BMB Reports 2022; 55(6): 275-280].
特应性皮炎(AD)的治疗具有挑战性,因为其病因复杂。从表皮破坏到慢性炎症,各种细胞和炎症途径都有助于 AD 的发展。与免疫抑制剂一样,普遍抑制炎症途径可能有效,但由于其副作用,这种方法不适合长期治疗。本研究旨在鉴定一种具有抗炎作用的植物提取物(PE),针对 AD 发展过程中涉及的多种细胞类型,并提供相关的机制证据。通过 RBL-2H3 细胞脱颗粒来筛选 30 种原产于韩国的 PE。为了研究朝鲜蓟耳状变种提取物(PAE)对 AD 的抗炎作用,使用 qRT-PCR、western blot、共聚焦显微镜、Griess 系统和 MTT 测定法在 RBL-2H3、HEK293、RAW264.7 和 HaCaT 细胞中评估细胞因子和一氧化氮的产生、FcεRI 和 TLR4 信号激活状态、细胞-细胞连接和细胞活力。对于体内实验,构建了 DNCB 诱导的 AD 小鼠模型,并进行了苏木精和伊红、过碘酸希夫、甲苯胺蓝和 F4/80 染色。通过 HPLC-MS 分析 PAE 的化学成分。通过测量 30 种 RBL-2H3 细胞中 PE 的抗脱颗粒作用,我们发现芍药、PA 和地黄(Gaertn.)Libosch. ex Steud. 的抑制活性超过 50%。其中,PAE 最显著和一致地抑制了细胞因子的表达,包括 IL-4、IL-9、IL-13 和 TNF-α。PAE 强烈抑制 FcεRI 信号,这从机制上支持其稳定嗜碱性粒细胞的作用,并且 PAE 通过干扰 Toll 样受体信号来下调巨噬细胞中的细胞因子和 NO 产生。此外,PAE 抑制角质形成细胞中细胞因子的产生,并上调紧密连接分子 ZO-1 和 occludin 的表达。在 DNCB 诱导的 AD 小鼠模型中,与地塞米松相比,PAE 的局部应用显著改善了特应性指数评分、免疫细胞浸润、细胞因子表达、FcεRI 和 TLR 信号中信号分子的异常激活以及受损的皮肤结构。PAE 的抗炎作用主要归因于 integerrimine。我们的研究结果表明,PAE 可以强烈抑制参与 AD 发展的多种炎症细胞,协同阻断炎症反应的传播,从而缓解 AD 症状。[BMB 报告 2022;55(6): 275-280]。