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槐定碱(来源于槐属植物)可改善实验性特应性皮炎小鼠模型。

Sophoricoside from Styphnolobium japonicum improves experimental atopic dermatitis in mice.

机构信息

Korea Institute of Science and Technology for Eastern Medicine (KISTEM), NEUMED Inc., Seoul 02440, Republic of Korea.

Department of Plant Science and Technology, Chung-Ang University, Anseong 17546, Republic of Korea.

出版信息

Phytomedicine. 2021 Feb;82:153463. doi: 10.1016/j.phymed.2021.153463. Epub 2021 Jan 14.

Abstract

BACKGROUND

Abnormal immune responses, specifically excessive differentiation of Th2 cells, are associated with the development of atopic dermatitis (AD). Sophoricoside, the genistein-4'-β-D-glucoside isolated from Styphnolobium japonicum, has previously demonstrated anti-inflammatory and immunosuppressive effects along with IL-3 and IL-5 inhibitory activities. Therefore, we speculated that sophoricoside could regulate AD by regulating abnormal immune responses.

PURPOSE

To investigate the role of sophoricoside on AD-like allergic skin inflammation induced by ovalbumin (OVA) or 2,4,6-trinitrochlorobenzene (TNCB) in mouse models.

METHODS

Sophoricoside was isolated from the 70% ethanol extract of S. japonicum dried mature seeds. After being submitted to a purification process, its purity was assessed by high-performance liquid chromatography (HPLC). The effects of sophoricoside were determined in vivo by OVA- and TNCB-induced AD-like allergic skin inflammation in mice. Skin tissues were subjected with hematoxylin-eosin (H&E), Giemsa and toluidine blue staining. In vitro CD4 T cell differentiation was performed and the levels of serum immunoglobulins, cytokines, and genes related to CD4 T cell differentiation were determined by enzyme-linked immunosorbent assay (ELISA) and quantitative real-time PCR. Cytokine bioassay, mixed lymphocytes reaction and cell viability assay were performed.

RESULTS

Topical application of sophoricoside decreased the symptoms of AD-like allergic skin inflammation, including elevated hypertrophic scars with spongiotic epidermis, epidermal hyperplasia, hyperkeratosis, infiltration of immune, and mast cells, dermal thickness, amounts of immunoglobulins, and pro-inflammatory cytokines, and the mast cell population in the skin. Sophoricoside also decreased T cell antigen receptor (TCR)-mediated immune responses. In particular, sophoricoside suppressed the differentiation of naïve CD4 T cells into Th cell subsets, including Th1, Th2, and Th17, by inhibiting the expression of their subset-specific master transcription factors, leading to suppression of the expression and production of these cell subset-specific cytokines.

CONCLUSION

Sophoricoside can improve AD-like allergic skin diseases mainly by inhibiting pathogenic CD4 T cell differentiation and immune responses.

摘要

背景

异常的免疫反应,特别是 Th2 细胞的过度分化,与特应性皮炎(AD)的发展有关。槐糖苷是从苦参中分离得到的染料木素-4'-β-D-葡萄糖苷,具有抗炎和免疫抑制作用,同时具有抑制 IL-3 和 IL-5 的活性。因此,我们推测槐糖苷可以通过调节异常免疫反应来调节 AD。

目的

研究槐糖苷对卵清蛋白(OVA)或 2,4,6-三硝基氯苯(TNCB)诱导的小鼠模型中 AD 样过敏性皮肤炎症的作用。

方法

槐糖苷从苦参干燥成熟种子的 70%乙醇提取物中分离得到。经纯化后,采用高效液相色谱法(HPLC)测定其纯度。通过 OVA 和 TNCB 诱导的 AD 样过敏性皮肤炎症的小鼠模型,观察槐糖苷的体内作用。对皮肤组织进行苏木精-伊红(H&E)、吉姆萨和甲苯胺蓝染色。进行 CD4 T 细胞分化的体外实验,并通过酶联免疫吸附测定(ELISA)和定量实时 PCR 测定血清免疫球蛋白、细胞因子和与 CD4 T 细胞分化相关的基因水平。进行细胞因子生物测定、混合淋巴细胞反应和细胞活力测定。

结果

槐糖苷的局部应用可减轻 AD 样过敏性皮肤炎症的症状,包括表皮海绵状肥厚、表皮增生、角化过度、免疫细胞和肥大细胞浸润、真皮增厚、免疫球蛋白和促炎细胞因子的含量以及皮肤中的肥大细胞数量。槐糖苷还可降低 T 细胞抗原受体(TCR)介导的免疫反应。特别是,槐糖苷通过抑制其亚群特异性主转录因子的表达,抑制幼稚 CD4 T 细胞向 Th 细胞亚群(包括 Th1、Th2 和 Th17)的分化,从而抑制这些细胞亚群特异性细胞因子的表达和产生。

结论

槐糖苷主要通过抑制致病 CD4 T 细胞分化和免疫反应来改善 AD 样过敏性皮肤病。

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