Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo
Abstract Background 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...
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| Format: | Article |
| Language: | English |
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BMC
2023-07-01
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| Series: | Journal of Translational Medicine |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12967-023-04293-2 |
| _version_ | 1827905309654384640 |
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| author | Yuqian Chang Pan Kang Tingting Cui Weinan Guo Weigang Zhang Pengran Du Xiuli Yi Sen Guo Tianwen Gao Chunying Li Shuli Li |
| author_facet | Yuqian Chang Pan Kang Tingting Cui Weinan Guo Weigang Zhang Pengran Du Xiuli Yi Sen Guo Tianwen Gao Chunying Li Shuli Li |
| author_sort | Yuqian Chang |
| collection | DOAJ |
| description | Abstract Background 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. Methods 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. Results 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. Conclusion 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. |
| first_indexed | 2024-03-13T00:39:51Z |
| format | Article |
| id | doaj.art-965a199f1d41414ca6d1b92f08574ed9 |
| institution | Directory Open Access Journal |
| issn | 1479-5876 |
| language | English |
| last_indexed | 2024-03-13T00:39:51Z |
| publishDate | 2023-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Translational Medicine |
| spelling | doaj.art-965a199f1d41414ca6d1b92f08574ed92023-07-09T11:23:07ZengBMCJournal of Translational Medicine1479-58762023-07-0121111910.1186/s12967-023-04293-2Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligoYuqian Chang0Pan Kang1Tingting Cui2Weinan Guo3Weigang Zhang4Pengran Du5Xiuli Yi6Sen Guo7Tianwen Gao8Chunying Li9Shuli Li10Department of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityDepartment of Dermatology, Fourth Military Medical UniversityAbstract Background 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. Methods 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. Results 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. Conclusion 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.https://doi.org/10.1186/s12967-023-04293-2DemethylzeylasteralVitiligoCD8+ T cellKeratinocytesJAKSTAT |
| spellingShingle | Yuqian Chang Pan Kang Tingting Cui Weinan Guo Weigang Zhang Pengran Du Xiuli Yi Sen Guo Tianwen Gao Chunying Li Shuli Li Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo Journal of Translational Medicine Demethylzeylasteral Vitiligo CD8+ T cell Keratinocytes JAK STAT |
| title | Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo |
| title_full | Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo |
| title_fullStr | Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo |
| title_full_unstemmed | Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo |
| title_short | Pharmacological inhibition of demethylzeylasteral on JAK-STAT signaling ameliorates vitiligo |
| title_sort | pharmacological inhibition of demethylzeylasteral on jak stat signaling ameliorates vitiligo |
| topic | Demethylzeylasteral Vitiligo CD8+ T cell Keratinocytes JAK STAT |
| url | https://doi.org/10.1186/s12967-023-04293-2 |
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