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复方生地合剂通过促进膜联蛋白 A 蛋白的表达缓解银屑病样皮肤炎症。

Fufang Shengdi mixture alleviates psoriasis-like skin inflammation via promoting Annexin-A proteins expression.

机构信息

Shanghai University of Traditional Chinese Medicine Science and Technology Experiment Center, Shanghai, 201203, China.

Shanghai University of Traditional Chinese Medicine School of Pharmacy, Shanghai, 201203, China.

出版信息

J Ethnopharmacol. 2023 Aug 10;312:116329. doi: 10.1016/j.jep.2023.116329. Epub 2023 Mar 20.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Traditional Chinese medicine believes that "blood fever" is an important cause of psoriasis. Fufang Shengdi mixture (FFSD), based on the Hongban Decoction, is composed of Rehmannia glutinosa (Gaertn.) DC., Raw gypsum (Chinese: Sheng Shi Gao), and Lonicera japonica Thunb (Caprifoliaceae). FFSD has effects on nourishing Yin, clearing heat, connecting collaterals, and cooling blood. In modern medical explanation, FFSD has the effects of anti-inflammatory and immunosuppression. Our study proved that FFSD can suppress immunity and ameliorate the symptoms of imiquimod-induced psoriasis in mice.

AIM OF THE STUDY

This study evaluated the efficacy and possible mechanism of FFSD in psoriasis mice.

METHODS AND MATERIALS

First, the main components of FFSD were analyzed using high-performance liquid chromatography-tandem high-resolution mass spectrometry (HPLC-HRMS). An imiquimod (IMQ)-induced psoriasis mouse model was used to evaluate the efficacy of FFSD orally. Psoriasis area and severity index (PASI) scores were recorded throughout the course of the mice to reflect the severity of psoriasis. Hematoxylin-eosin staining was used to observe the pathological changes in skin lesions. Enzyme-linked immunosorbent assay (ELISA) was performed to test the level of IFN-γ and TNF-α in plasma. To further investigate the immunopharmacological effect of FFSD, we used chicken ovalbumin (OVA) to induce immunoreaction in mice. ELISA was used to detect the levels of anti-OVA antibody, IFN-γ and TNF-α in mice. Flow cytometry was performed to quantify the ratio of cell types in peripheral blood mononuclear cells (PBMCs) to evaluate the effect of FFSD on immunosuppression. Proteomics and bioinformatics analyzes were performed to find the regulation pathway of the immunosuppressive effect of FFSD. Finally, quantitative PCR (qPCR) and immunohistochemistry were used to measure the upregulation of Annexin-A proteins (ANXAs) in the skin lesion tissue of IMQ-induced mouse.

RESULTS

On the basis of knowing the composition of FFSD, we first proved the efficacy of FFSD in alleviating IMQ-induced psoriasis in mice. Second, we further clarified the pharmacological effect of FFSD on immunosuppression via OVA-induced mice. Subsequently, it was found that the significant up-regulation of ANXAs was caused by FFSD through proteomics analysis, and the finding was proved in the IMQ-induced psoriasis mouse model.

CONCLUSIONS

This study elucidates the immunosuppressive pharmacological effect of FFSD on improving psoriasis through up-regulating ANXAs.

摘要

民族药理学相关性

中医认为“血热”是银屑病的重要病因。复方生地合剂(FFSD)基于红芪汤,由地黄(Gaertn.)、生石膏和金银花(忍冬科)组成。FFSD 具有滋阴清热、通络凉血的作用。在现代医学解释中,FFSD 具有抗炎和免疫抑制作用。我们的研究证明,FFSD 可抑制免疫,改善咪喹莫特诱导的银屑病小鼠的症状。

研究目的

本研究评价 FFSD 对银屑病小鼠的疗效及可能机制。

方法和材料

首先,采用高效液相色谱-串联高分辨率质谱法(HPLC-HRMS)分析 FFSD 的主要成分。采用咪喹莫特(IMQ)诱导的银屑病小鼠模型评价 FFSD 的疗效。在整个小鼠过程中记录银屑病面积和严重程度指数(PASI)评分,以反映银屑病的严重程度。苏木精-伊红染色观察皮肤病变的病理变化。酶联免疫吸附试验(ELISA)检测血浆中 IFN-γ和 TNF-α的水平。为进一步研究 FFSD 的免疫药理学作用,我们用鸡卵清蛋白(OVA)诱导小鼠免疫反应。ELISA 检测小鼠抗 OVA 抗体、IFN-γ和 TNF-α水平。流式细胞术检测外周血单个核细胞(PBMCs)中细胞类型的比例,评价 FFSD 对免疫抑制的作用。蛋白质组学和生物信息学分析寻找 FFSD 免疫抑制作用的调控途径。最后,采用定量 PCR(qPCR)和免疫组织化学法检测 IMQ 诱导的小鼠皮肤病变组织中 Annexin-A 蛋白(ANXAs)的上调。

结果

在了解 FFSD 组成的基础上,我们首先证明了 FFSD 缓解 IMQ 诱导的银屑病的疗效。其次,我们通过 OVA 诱导的小鼠进一步阐明了 FFSD 对免疫抑制的药理作用。随后,通过蛋白质组学分析发现 FFSD 引起的 ANXAs 显著上调,并在 IMQ 诱导的银屑病小鼠模型中得到证实。

结论

本研究通过上调 ANXAs 阐明了 FFSD 改善银屑病的免疫抑制药理作用。

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