Zhang Yijian, Wang Xueting, Yang Yang, Zhao Long, Tu Huiyang, Zhang Yiyu, Hu Guoliang, Tian Chong, Zhang Beibei, Bai Zhaofang, Zhang Bin
Anhui Medical University PLA 307 Clinical College, Beijing 100071; Senior Department of Hematology, the Fifth Medical Center of PLA General Hospital, Beijing 100071; Beijing Key Laboratory of Hematopoietic Stem Cell Therapy and Transformation Research, Beijing 100071; The Fifth Clinical Medical College of Anhui Medical University, Hefei 230032, China.
Senior Department of Hematology, the Fifth Medical Center of PLA General Hospital, Beijing 100071; Beijing Key Laboratory of Hematopoietic Stem Cell Therapy and Transformation Research, Beijing 100071; The School of Medicine, Nankai University, Tianjin 300071, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2025 May;41(5):428-436.
Objective To investigate the effects of cucurbitacin B (CucB) on alleviating skin lesions and inflammation in psoriasis mice via the cGAS-STING signaling pathway. Methods The expression of genes associated with the cGAS-STING signaling pathway in psoriatic lesions and non-lesional skin was analyzed, and hallmark gene set enrichment analysis was performed. The cytotoxicity of CucB on BMDMs was evaluated using the CCK-8 assay. The expression levels of genes and proteins related to the cGAS-STING signaling pathway, along with the secretion of inflammatory cytokines, were measured at different concentrations of CucB using quantitative PCR, Western blotting, and ELISA. Imiquimod-induced psoriasis BALB/c mice were divided into four groups: normal group, model group, low-dose CucB group [0.1 mg/ (kg.d)], and high-dose CucB group [0.4 mg/ (kg.d)], with five mice per group. PASI scoring was performed to assess the severity of psoriasis after 6 days of treatment, and HE staining was conducted to observe pathological damage. Meanwhile, the mRNA levels of inflammatory cytokines and their secretion were detected by qPCR and ELISA. Results Most cGAS-STING signaling-related genes were upregulated in lesional skin of psoriasis patients, and the hallmark gene set enrichment analysis revealed that the most significantly upregulated genes were primarily associated with immune response signaling pathways. CucB inhibited dsDNA-induced phosphorylation of interferon regulatory factor 3 (IRF3) and STING proteins in both bone-marrow derived macrophages(BMDMs) and THP-1 cells. CucB also suppressed dsDNA-induced mRNA expression of IFNB1, TNF, IFIT1, CXCL10, ISG15, and reduced the secretion of cytokines such as IFN-β, IL-1β, and TNF-α in THP-1 cells. In the imiquimod-induced psoriasis mouse model, CucB treatment reduced psoriatic symptoms, alleviated skin lesions, and attenuated inflammation. ELISA and qPCR results showed that CucB significantly reduced serum secretion levels of IL-6, TNF-α, and IL-1β, as well as the mRNA levels of IL23A, IL1B, IL6, TNF, and IFNB1. Conclusion CucB inhibits cytoplasmic DNA-induced activationc of the GAS-STING pathway. CucB significantly attenuates skin lesions and inflammation in IMQ-induced psoriatic mice, and the potential molecular mechanism may be related to the down-regulation of the cGAS-STING pathway.
目的 探讨葫芦素B（CucB）通过环鸟苷酸-腺苷酸合成酶-干扰素基因刺激蛋白（cGAS-STING）信号通路减轻银屑病小鼠皮肤损伤和炎症的作用。方法 分析银屑病皮损及非皮损皮肤中与cGAS-STING信号通路相关基因的表达，并进行特征基因集富集分析。采用细胞计数试剂盒-8（CCK-8）法评估CucB对骨髓来源巨噬细胞（BMDMs）的细胞毒性。使用定量聚合酶链反应（PCR）、蛋白质免疫印迹法（Western blotting）和酶联免疫吸附测定（ELISA）检测不同浓度CucB作用下与cGAS-STING信号通路相关的基因和蛋白表达水平以及炎性细胞因子的分泌情况。将咪喹莫特诱导的银屑病BALB/c小鼠分为四组：正常组、模型组、低剂量CucB组[0.1 mg/（kg·d）]和高剂量CucB组[0.4 mg/（kg·d）]，每组5只小鼠。治疗6天后进行银屑病面积和严重程度指数（PASI）评分以评估银屑病严重程度，并进行苏木精-伊红（HE）染色观察病理损伤。同时，通过qPCR和ELISA检测炎性细胞因子的mRNA水平及其分泌情况。结果 大多数与cGAS-STING信号相关的基因在银屑病患者的皮损皮肤中上调，特征基因集富集分析显示上调最显著的基因主要与免疫反应信号通路相关。CucB抑制双链DNA（dsDNA）诱导的骨髓来源巨噬细胞（BMDMs）和人单核细胞白血病细胞系（THP-1）中干扰素调节因子3（IRF3）和STING蛋白的磷酸化。CucB还抑制dsDNA诱导的THP-1细胞中IFNB1、TNF、IFIT1、CXCL10、ISG15的mRNA表达，并减少细胞因子如干扰素-β（IFN-β）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）的分泌。在咪喹莫特诱导的银屑病小鼠模型中，CucB治疗减轻了银屑病症状，缓解了皮肤损伤并减轻了炎症。ELISA和qPCR结果显示，CucB显著降低了白细胞介素-6（IL-6）、TNF-α和IL-1β的血清分泌水平，以及IL23A、IL1B、IL6、TNF和IFNB1的mRNA水平。结论 CucB抑制细胞质DNA诱导的cGAS-STING通路激活。CucB显著减轻咪喹莫特诱导的银屑病小鼠的皮肤损伤和炎症，其潜在分子机制可能与下调cGAS-STING通路有关。
