Abstract
T helper (Th) 17 cells highly contribute to the immunopathology of rheumatoid arthritis. Morin, a natural flavonoid, owns well anti-arthritic action but unclear effect on Th17 differentiation. This study tried to solve this issue and explore the mechanisms in view of cellular metabolism. Naïve CD4+ T cells were treated with anti-CD3/CD28 along with Th17-inducing cytokines. Morin was shown to block Th17 differentiation without affecting cell viability even when Foxp3 was dampened. The mechanisms were ascribed to the limited fatty acid synthesis by restricting FASN transcription, as indicated by metabolomics analysis, nile red staining, detection of triglycerides, FASN overexpression, and addition of palmitic acid. Moreover, morin had slight effect on cell apoptosis and protein palmitoylation during Th17 differentiation, but blocked the binding of RORγt to promoter and CNS2 region of Il17a gene. Oleic acid rescued the inhibition of morin on RORγt function, and Th17-inducing cytokines could not induce RORγt function in SCD1-defficient cells, suggesting that oleic acid but not palmitic acid was the direct effector in the action of morin. Then, PPARγ was identified as the target of morin, and GW9662 or PPARγ CRISPR/Cas9 KO plasmid weakened its above-mentioned effects. The transrepression of FASN by morin was owing to physical interaction between PPARγ and Sp1, and the importance of Sp1 in Th17 differentiation was confirmed by siSp1. Finally, the effects and mechanisms for morin-dampened Th17 responses were confirmed in collagen-induced arthritis (CIA) mice. Collectively, morin inhibited Th17 differentiation and alleviated CIA by limiting fatty acid synthesis subsequent to PPARγ activation.
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Funding
This work was supported by the National Natural Science Foundation of China (81773970), Qing Lan Project of Jiangsu Province (2019), the “Double First-Class” University Project (CPU2018GY10), the University Innovation Research and Training Program of China Pharmaceutical University (202110316068), and partially supported by the Priority Academic Program Development of Jiangsu Higher Education Institutions.
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Zhifeng Wei and Yue Dai designed the study. Yumeng Miao, Xiaoqian Wu, Xinru Xue, Xingyu Ma, Ling Yang, Xi Zeng, and Yuxiao Hu performed the experiments and statistical analysis. Yumeng Miao additionally contributed to the preparation of the manuscript. All of the authors approved the final version of this manuscript.
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This study was approved by the Animal Ethics Committee of China Pharmaceutical University and followed the National Institute of Health guidelines on the ethical use of animals.
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Graphical headlights
1. Morin limits Th17 differentiation by restricting FASN-mediated fatty acid synthesis
2. Morin dampens RORγt function subsequent to the restriction of fatty acid synthesis
3. Morin inhibits FASN transcription via PPARγ-Sp1 signaling
4. Morin carries out anti-arthritic action through limiting Th17 responses
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Miao, Y., Wu, X., Xue, X. et al. Morin, the PPARγ agonist, inhibits Th17 differentiation by limiting fatty acid synthesis in collagen-induced arthritis. Cell Biol Toxicol 39, 1433–1452 (2023). https://doi.org/10.1007/s10565-022-09769-3
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DOI: https://doi.org/10.1007/s10565-022-09769-3