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4-Octyl Itaconate Inhibits Proinflammatory Cytokine Production in Behcet’s Uveitis and Experimental Autoimmune Uveitis

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Abstract

4-octyl itaconate (4-OI) is an anti-inflammatory metabolite that activates the nuclear-factor-E2-related factor 2 (NRF2) signaling. In the current work, we investigated whether 4-OI could affect the production of proinflammatory cytokines in Behcet’s uveitis (BU) and experimental autoimmune uveitis (EAU). Peripheral blood mononuclear cells (PBMCs) of active BU patients and healthy individuals with in vitro 4-OI treatment were performed to assess the influence of 4-OI on the proinflammatory cytokine production. EAU was induced and used for investigating the influence of 4-OI on the proinflammatory cytokine production in vivo. The flow cytometry, qPCR, and ELISA were performed to detect proinflammatory cytokine expression. NRF2 signaling activation was evaluated by qPCR and western blotting (WB). Splenic lymphocyte transcriptome was performed by RNA sequencing. The NRF2 expression by BU patients-derived PBMCs was lower than that by healthy individuals. After treatment with 4-OI, the proportion of Th17 cells, along with the expression of proinflammatory cytokines (IL-17, TNF-α, MCP-1, and IL-6) by PBMCs, were downregulated, and anti-inflammatory cytokine (IL-10) expression was upregulated, although IFN-γ expression was unaffected. The EAU severity was ameliorated by 4-OI in association with a lower splenic Th1/Th17 cell proportion and increased nuclear NRF2 expression. Additionally, 4-OI downregulated a set of 248 genes, which were enriched in pathways of positive regulation of immune responses. The present study shows an inhibitory effect of 4-OI on the proinflammatory cytokine production in active BU patients and EAU mice, possibly mediated through activating NRF2 signaling. These findings suggest that 4-OI could act as a potential therapeutic drug for the treatment and prevention of BU in the future study.

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Availability of Data and Material

The data sets used and/or analyzed in this study are available from the corresponding authors upon reasonable request. The dataset of RNA-seq is available in https://www.jianguoyun.com/, and the shared data link is https://www.jianguoyun.com/p/DRZEH24Qn5H8CxjW55sFIAA.

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Acknowledgements

We appreciate the technical support provided by our laboratory’s entire clinical research personnel and students. We also appreciate the cooperation and informed consent provided by the healthy donors and patients for this study.

Funding

This study was supported by the National Natural Science Foundation Key Program (82230032), National Natural Science Foundation Key Program (81930023), Natural Science Foundation Major International (Regional) Joint Research Project (81720108009), Key Project of Chongqing Science and Technology Bureau (CSTC2021jscx-gksb-N0010), National Natural Science Foundation (82101106), Foundation of Chongqing (cstc2021jcyj-bshX0154), Chongqing Outstanding Scientists Project (2019), Chongqing Chief Medical Scientist Project (2018), Chongqing Key Laboratory of Ophthalmology (CSTC, 2008CA5003), and Chongqing Science & Technology Platform and Base Construction Program (cstc2014pt-sy10002).

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  1. Qingfeng Wang and Xingsheng Ye contributed equally to this work. 

Authors and Affiliations

  1. The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, Chongqing Branch (Municipality Division) of National Clinical Research Center for Ocular Diseases, Chongqing, People’s Republic of China

    Qingfeng Wang, Xingsheng Ye, Shiyao Tan, Qingyan Jiang, Guannan Su, Su Pan, Hongxi Li, Qingfeng Cao & Peizeng Yang

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  1. Qingfeng Wang
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Contributions

QW and PY: conceived and directed the study. QW, XY, ST, HL, QJ, and SP: analyzed the data. PY: made the clinical diagnoses. QC: collected the samples. QW, XY, ST, and HL: performed the in vitro experiment and animal experiment. QW and XY: evaluated histology slides. QW and GS: drafted the manuscript. PY: reviewed the data interpretation and helped revise the final versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Peizeng Yang.

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Ethics Approval and Consent to Participate

This study was approved by the Ethics Committee of Chongqing Medical University (2018–048). All procedures were carried out in accordance with the Declaration of Helsinki. All participants signed an informed consent form at the start of the study. The animal experiments were approved by the Ethics Committee of the First Affiliated Hospital of Chongqing Medical University.

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The authors declare no competing interests.

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Supplementary Information

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Supplementary file1 (DOCX 35 KB)

10753_2023_1950_MOESM2_ESM.tif

Supplementary file2: The effects of 4-OI on the Th1/Th17 index cytokines as well as proinflammatory and anti-inflammatory cytokines mRNA expression by retinas of EAU mice. (A) Comparison of IFN-γ and IL-17 mRNA expression between 4-OI-treated group and control group. (B) Comparison of IL-6, TNF-α, MCP-1and IL-10 mRNA expression between 4-OI-treated group and control group. β-actin was used as the internal reference. Fold change is used to show the gene expression in the 4-OI-treated group relative to that in the control group. Data were represented as the relative fold change (2−∆∆Ct). Data were analyzed by unpaired t-test (n = 7 per group). *p < 0.05; **p < 0.01. (TIF 106 KB)

10753_2023_1950_MOESM3_ESM.tif

Supplementary file3: Validation of five DEGs from the RNA-seq data. β-actin was used as the internal reference. Fold change is used to show the gene expression in the 4-OI-treated group relative to that in the control group. Data were represented as the relative fold change (2−∆∆Ct). Data were analyzed by unpaired t-test (n = 7 per group). *p < 0.05; **p < 0.01. (TIF 90 KB)

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Wang, Q., Ye, X., Tan, S. et al. 4-Octyl Itaconate Inhibits Proinflammatory Cytokine Production in Behcet’s Uveitis and Experimental Autoimmune Uveitis. Inflammation (2024). https://doi.org/10.1007/s10753-023-01950-y

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  • DOI: https://doi.org/10.1007/s10753-023-01950-y

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