Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo.
Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo.
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去甲基泽兰醛对JAK-STAT信号的药理抑制可改善白癜风。
DOI：
10.1186/s12967-023-04293-2
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发表时间：
2023-07-04
影响因子：
7.4
通讯作者：
Li, Shuli
中科院分区：
文献类型：
--
作者：
Chang, Yuqian;Kang, Pan;Cui, Tingting;Guo, Weinan;Zhang, Weigang;Du, Pengran;Yi, Xiuli;Guo, Sen;Gao, Tianwen;Li, Chunying;Li, Shuli
CD8+ T细胞的激活及其通过JAK-STAT信号向皮肤的运输在白癜风的发展中起着核心作用。因此，利用创新药物靶向这一关键疾病通路是治疗白癜风的有效策略。从草药中分离的天然产物是新疗法的有用来源。从雷公藤中提取的去甲基zeylastal （T-96）具有免疫抑制和抗炎作用。在小鼠白癜风模型中检测T-96的作用，并采用全挂载尾染色法定量观察表皮中CD8+ T细胞的浸润和黑色素细胞的数量。采用流式细胞术评价T-96在CD8+ T细胞中的免疫调节作用。采用拉下分析、质谱分析、分子对接、敲低和过表达等方法鉴定T-96在CD8+ T细胞和角质形成细胞中的靶蛋白。在我们的白癜风小鼠模型中，我们发现T-96通过全挂尾染色减少了表皮中CD8+ T细胞的浸润，并减轻了与托法替尼（Tofa）相当程度的色素脱色。在体外，T-96可降低白癜风患者CD8+ T细胞的增殖、CD69膜表达以及IFN-γ、颗粒酶B （GzmB）和穿孔素（PRF）水平。Pull-down实验结合质谱分析和分子对接表明，T-96在CD8+ T细胞裂解物中与JAK3相互作用。此外，在IL-2处理后，T-96降低了JAK3和STAT5的磷酸化。T-96不能进一步降低JAK3敲低后IFN-γ、GzmB和PRF的表达，也不能抑制JAK3过表达后免疫效应物的表达。此外，T-96在IFN-γ刺激的角质形成细胞中与JAK2相互作用，抑制JAK2的激活，降低STAT1的总蛋白和磷酸化蛋白水平，减少CXCL9和CXCL10的产生和分泌。T-96在JAK2下调后没有显著抑制STAT1和CXCL9/10的表达，也没有抑制JAK2过表达后STAT1-CXCL9/10信号的上调。最后，T-96降低了CXCR3的膜表达，在IFN-γ应激的角质形成细胞下，用T-96预处理的培养上清明显阻断了CXCR3+CD8+ T细胞的迁移，与体外的Tofa相似。我们的研究结果表明，T-96可能通过JAK-STAT信号传导抑制CD8+ T细胞的效应功能和皮肤运输，从而对白癜风具有积极的治疗作用。在线版本包含补充材料，可在10.1186/s12967-023-04293-2获得。
The activation of CD8+ T cells and their trafficking to the skin through JAK-STAT signaling play a central role in the development of vitiligo. Thus, targeting this key disease pathway with innovative drugs is an effective strategy for treating vitiligo. Natural products isolated from medicinal herbs are a useful source of novel therapeutics. Demethylzeylasteral (T-96), extracted from Tripterygium wilfordii Hook F, possesses immunosuppressive and anti-inflammatory properties. The efficacy of T-96 was tested in our mouse model of vitiligo, and the numbers of CD8+ T cells infiltration and melanocytes remaining in the epidermis were quantified using whole-mount tail staining. Immune regulation of T-96 in CD8+ T cells was evaluated using flow cytometry. Pull-down assay, mass spectrum analysis, molecular docking, knockdown and overexpression approaches were utilized to identify the target proteins of T-96 in CD8+ T cells and keratinocytes. Here, we found that T-96 reduced CD8+ T cell infiltration in the epidermis using whole-mount tail staining and alleviated the extent of depigmentation to a comparable degree of tofacitinib (Tofa) in our vitiligo mouse model. In vitro, T-96 decreased the proliferation, CD69 membrane expression, and IFN-γ, granzyme B, (GzmB), and perforin (PRF) levels in CD8+ T cells isolated from patients with vitiligo. Pull-down assays combined with mass spectrum analysis and molecular docking showed that T-96 interacted with JAK3 in CD8+ T cell lysates. Furthermore, T-96 reduced JAK3 and STAT5 phosphorylation following IL-2 treatment. T-96 could not further reduce IFN-γ, GzmB and PRF expression following JAK3 knockdown or inhibit increased immune effectors expression upon JAK3 overexpression. Additionally, T-96 interacted with JAK2 in IFN-γ-stimulated keratinocytes, inhibiting the activation of JAK2, decreasing the total and phosphorylated protein levels of STAT1, and reducing the production and secretion of CXCL9 and CXCL10. T-96 did not significantly inhibit STAT1 and CXCL9/10 expression following JAK2 knockdown, nor did it suppress upregulated STAT1-CXCL9/10 signaling upon JAK2 overexpression. Finally, T-96 reduced the membrane expression of CXCR3, and the culture supernatants pretreated with T-96 under IFN-γ stressed keratinocytes markedly blocked the migration of CXCR3+CD8+ T cells, similarly to Tofa in vitro. Our findings demonstrated that T-96 might have positive therapeutic responses to vitiligo by pharmacologically inhibiting the effector functions and skin trafficking of CD8+ T cells through JAK-STAT signaling. The online version contains supplementary material available at 10.1186/s12967-023-04293-2.
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影响因子：
5.6
作者：
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通讯作者：
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DOI：
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影响因子：
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作者：
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影响因子：
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DOI：
10.4049/jimmunol.2000612
发表时间：
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期刊：
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通讯作者：
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