BML-111 调节和缓解小鼠银屑病样皮炎中的 p38/MAPK 信号通路和 Th1/Th2/Th17 细胞因子反应。

BML-111 Modulates and Alleviates p38/MAPK Signaling Pathway and Th1/Th2/Th17 Cytokine Response in Murine Psoriasis-Like Dermatitis.

机构信息

Department of Dermatology, The First Affiliated Hospital of Heilongjiang University of Chinese Medicine, 150040 Harbin, Heilongjiang, China.

出版信息

Discov Med. 2024 Oct;36(189):2026-2036. doi: 10.24976/Discov.Med.202436189.186.

DOI:10.24976/Discov.Med.202436189.186

PMID:39463222

Abstract

BACKGROUND

Psoriasis is a prevalent cutaneous inflammatory disorder characterized by elevated keratinocyte inflammation. 5(S)-6(R)-7-trihydroxyheptanoic-acid-methyl-ester (BML-111), an established analogue of lipoxin A4, is known for its potent anti-inflammatory properties. However, the precise role of BML-111 within a murine psoriasis-like dermatitis model requires further clarification. This research aims to investigate the modulatory effects of BML-111 on inflammatory responses, the p38/mitogen-activated protein kinase (MAPK) signaling cascade, and T helper type 1 (Th1), Th2, and Th17 cell responses within the context of a murine psoriasis-like dermatitis model.

METHODS

A psoriasis-like dermatitis model was established by applying 5% imiquimod (IMQ) cream to the backs of C57BL/6 mice, which were pretreated intraperitoneally with or without BML-111 prior to IMQ application. Hematoxylin-eosin staining was utilized to detect the pathological alterations of the murine dorsal skin tissue. Furthermore, the psoriasis area and severity index (PASI) scoring system was used to assess the dynamic cutaneous alterations in the mice. The levels of tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), interleukin (IL)-1β, IL-6, IL-4, and IL-17A in the murine serum samples were quantified by means of enzyme-linked immunosorbent assays (ELISA). Western blotting was conducted to detect the proteins of TNF-α, IL-1β, IL-6, phospho-p38 (p-p38), and p38 in murine skin tissues. Lastly, a flow cytometry analysis was executed to evaluate the expression of peripheral blood Th1/Th2/Th17 cell subsets.

RESULTS

BML-111 attenuated IMQ-induced pathological changes in skin tissue of psoriasis-like dermatitis mice. BML-111 treatment substantially reduced TNF-α, IL-1β, IL-6, IFN-γ and IL-17A levels and elevated IL-4 levels in serum and skin lesion tissues of IMQ-induced mice ( < 0.01, < 0.01, < 0.01, < 0.05, < 0.05, < 0.05, respectively). The ratio of Th1/Th17 cells in the peripheral blood of BML-111-treated mice was substantially diminished and the ratio of Th2 cells was substantially augmented ( < 0.05, < 0.01, < 0.001, respectively). Mechanistically, p-p38 protein level was substantially reduced in the skin tissues of BML-111-treated mice ( < 0.05). While, dehydrocorydaline (DHC, a p38/MAPK pathway agonists) reversed the reduction of p-p38 protein level induced by BML-111 treatment in psoriasis-like mice ( < 0.05).

CONCLUSION

BML-111 modulates the p38/MAPK signaling pathway and Th1/Th2/Th17 cytokine response, and alleviates psoriasis-like dermatitis in mice.

摘要

背景

银屑病是一种常见的皮肤炎症性疾病，其特征为角质形成细胞炎症升高。5(S)-6(R)-7-三羟基庚酸甲酯（BML-111）是脂氧素 A4 的一种成熟类似物，以其强大的抗炎特性而闻名。然而，BML-111 在类似银屑病的皮肤炎模型中的精确作用仍需要进一步阐明。本研究旨在研究 BML-111 对炎症反应、p38/丝裂原激活蛋白激酶（MAPK）信号级联和 Th1、Th2 和 Th17 细胞反应的调节作用，在类似银屑病的皮肤炎模型中。

方法

通过在 C57BL/6 小鼠背部涂抹 5%咪喹莫特（IMQ）乳膏，建立类似银屑病的皮肤炎模型，在应用 IMQ 前，通过腹腔内预先给予或不给予 BML-111 对其进行预处理。通过苏木精-伊红染色检测小鼠背部皮肤组织的病理改变。此外，采用银屑病面积和严重程度指数（PASI）评分系统评估小鼠皮肤的动态变化。通过酶联免疫吸附试验（ELISA）定量检测小鼠血清中肿瘤坏死因子-α（TNF-α）、干扰素-γ（IFN-γ）、白细胞介素（IL）-1β、IL-6、IL-4 和 IL-17A 的水平。采用 Western blot 检测小鼠皮肤组织中 TNF-α、IL-1β、IL-6、磷酸化 p38（p-p38）和 p38 的蛋白。最后，通过流式细胞术分析评估外周血 Th1/Th2/Th17 细胞亚群的表达。

结果

BML-111 减轻了 IMQ 诱导的类似银屑病的皮肤炎小鼠皮肤组织的病理变化。BML-111 治疗可显著降低 IMQ 诱导的小鼠血清和皮肤损伤组织中 TNF-α、IL-1β、IL-6、IFN-γ 和 IL-17A 的水平，并升高 IL-4 的水平（<0.01，<0.01，<0.01，<0.05，<0.05，<0.05，分别）。BML-111 治疗后，小鼠外周血中 Th1/Th17 细胞的比例显著降低，Th2 细胞的比例显著升高（<0.05，<0.01，<0.001，分别）。BML-111 治疗小鼠皮肤组织中 p-p38 蛋白水平显著降低（<0.05）。而脱氢紫堇碱（DHC，p38/MAPK 通路激动剂）可逆转 BML-111 治疗引起的 p-p38 蛋白水平降低（<0.05）。