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Protective Effect of Saffron in Mouse Colitis Models Through Immune Modulation

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Abstract

Background

People with inflammatory bowel disease (IBD) including ulcerative colitis are at risk for colorectal cancer. Despite available effective drugs used to treat IBD, many patients fail or lose response over time with some displaying drug-induced adverse events. Saffron (Crocus sativus) has been reported to have anti-inflammatory properties. Its protective role in IBD has not been explored extensively.

Aim

To establish whether saffron treatment alleviates inflammation in experimental colitis.

Methods

Colitis was induced in C57BL/6 mice with 3% DSS and treated with either saffron doses (7.5, 15, 20, 25 mg/kg body weight) or vehicle through daily gavage. On day 11, mice were euthanized and analyzed for gross and microscopic inflammation. Distal colon segments were collected for mRNA and protein expression of HO-1 protein and GPX2, (the downstream targets of NRF-2). Nrf-2 translocation from cytosol to nucleus was confirmed by immunofluorescence, and further Nrf-2 protein expression in nuclear and cytosolic fraction of colon was analyzed by immunoblot. Immune cells were isolated from the lamina propria of mouse colon for flow cytometry-based immunophenotyping. Colitis was also induced in C57BL/6 Ahr knockout and wild type mice to explore the involvement of Ahr-dependent pathways in saffron’s protective effect(s). The therapeutic effect of saffron was further validated in another TNBS model of colitis.

Results

Saffron 20 mg/kg body weight showed improved colon gross and histology features and led to better body weight, colon length, histology score, and reduced disease activity index (DAI). Saffron significantly decreased pro-inflammatory macrophages (M1), while increasing anti-inflammatory macrophages (M2) and IL10 + dendritic cells. Saffron treatment also enhanced CD3 + T and CD3 + CD8 + T cells followed by increase in different CD3 + CD4 + T cells subsets like CD25 + T cells, FoxP3 + CD25 + regulatory T cells, and CD4 + FOXP3 + CD25-regulatory T cells. Immunoblot analysis showed a significant increase in HO-1/GPX2 protein expression. With saffron treatment, Nrf-2 translocation into nucleus from cytosol also supports the involvement of Nrf-2 and its downstream targets in the protective effect of saffron. Further, we demonstrated that saffron in part exert anti-inflammatory effect through activation of aryl hydrocarbon receptor (AhR)-nuclear factor erythroid 2–related factor 2 (Nrf2)-dependent pathways.

Conclusion

These data demonstrate saffron’s therapeutic potential and its protective role in part via Ahr/Nrf-2 pathways and regulatory innate and adaptive immune cells.

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Abbreviations

IBD:

Inflammatory bowel disease

DSS:

Dextran sodium sulfate

NRF-2:

Nuclear factor erythroid 2

Ahr:

Aryl hydrocarbon receptor

HO-1:

Hemeoxygenase-1

GPX2:

Glutathione peroxidase 2

TNF-α:

Tumor necrosis factor-alpha

DAI:

Disease activity index

DCs:

Dendritic cells

CD:

Crohn’s disease

UC:

Ulcerative colitis

SFE:

Saffron aqueous extract

TNBS:

Trinitrobenzenesulfonic acid

HBSS:

Hank's balanced salt solution (HBSS)

BCS:

Bovine calf serum

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Acknowledgments

We would like to thank Gulf Pearls SPRL (Brussels, Belgium, www.gp-food.com) that provided saffron for our research studies. In addition, we would like to thank all the saffron growers and people involved in the production of this golden spice.

Funding

This project was supported (in part) by the National Institute on Minority Health and Health Disparities of the National Institutes of Health under Award Number G12MD007597.

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Author notes

  1. Aida Habtezion, Hassan Ashktorab have contributed equally to this work.

Authors and Affiliations

  1. Gastroenterology Division and Hepatology, School of Medicine, Stanford University, Palo Alto, CA, USA

    Gulshan Singh, Yeneneh Haileselassie, Allison Ruoheng Ji, Sidhartha R. Sinha & Aida Habtezion

  2. Microbiology & Immunology, Human Immune Monitoring Center, Stanford University, Palo Alto, CA, USA

    Holden Terry Maecker

  3. Department of Medicine and Cancer Center, Howard University College of Medicine, Washington, DC, USA

    Hassan Ashktorab

  4. Department of Pathology, Howard University College of Medicine, Washington, DC, USA

    Hassan Brim

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  1. Gulshan Singh
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Correspondence to Hassan Ashktorab.

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Singh, G., Haileselassie, Y., Ji, A.R. et al. Protective Effect of Saffron in Mouse Colitis Models Through Immune Modulation. Dig Dis Sci 67, 2922–2935 (2022). https://doi.org/10.1007/s10620-021-07163-3

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