Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Nov 15;27(22):7871.
doi: 10.3390/molecules27227871.

Immunomodulatory Activity of Punicalagin, Punicalin, and Ellagic Acid Differs from the Effect of Pomegranate Peel Extract

Miodrag Čolić  1   2 , Dušan Mihajlović  1 , Marina Bekić  3 , Milan Marković  3 , Branka Dragišić  4 , Sergej Tomić  3 , Nataša Miljuš  4 , Katarina Šavikin  5 , Ranko Škrbić  4
Affiliations

Immunomodulatory Activity of Punicalagin, Punicalin, and Ellagic Acid Differs from the Effect of Pomegranate Peel Extract

Miodrag Čolić et al. Molecules. .

Abstract

Background: Our recent study has shown that pomegranate peel extract (PEx) showed significant immunomodulatory activity, which might be caused by ellagitannins. The aim of this work was to test the hypothesis that ellagitannin components act synergistically in the modulation of cytokine production.

Methods: Human peripheral blood mononuclear cells (PBMCs) from healthy donors were stimulated with phytohemagglutinin and treated with different concentrations of PEx or punicalagin (PG), punicalin (PN), and ellagic acid (EA), alone or with their combinations. Cytotoxicity, cell proliferation, and cytokine production were determined.

Results: Non-cytotoxic concentrations of all compounds significantly inhibited cell proliferation. IC50 values (μg/mL) were: EA (7.56), PG (38.52), PEx (49.05), and PN (69.95). PEx and all ellagitannins inhibited the levels of TNF-α, IL-6, and IL-8, dose-dependently, and their combinations acted synergistically. PEx and all ellagitannins inhibited Th1 and Th17 responses, whereas the lower concentrations of PEx stimulated the production of IL-10, a Treg cytokine, as did lower concentrations of EA. However, neither component of ellagitannins increased Th2 response, as was observed with PEx.

Conclusions: The combination of PG, PN, and EA potentiated the anti-inflammatory response without any significant synergistic down-modulatory effect on T-cell cytokines. The increased production of IL-10 observed with PEx could be attributable to EA, but the examined ellagitannins are not associated with the stimulatory effect of PEx on Th2 response.

Keywords: culture; cytokines; ellagitannins; lymphocytes; pomegranate.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Cytotoxicity of pomegranate peel extract (PEx), punicalagin (PG), punicalin (PN) and ellagic acid (EA) in culture with PBMC. Cytotoxicity of PBMC was determined after 24 using the MTT test as described in Materials and methods. Values are shown as mean ± SD of five donors. *** p < 0.005, compared to control non-treated PBMC.
Figure 2
Figure 2
The effects of pomegranate peel extract (PEx), punicalagin (PG), punicalin (PN) and ellagic acid (EA) on PHA-stimulated proliferation of PBMC. PBMC pre-labeled with Cell Trace Far-Red were cultured with different concentrations of the compounds (1–320 μg/mL) or without them in the presence of PHA (10 μg/mL) for 4 days, followed by the analysis of Far-Red dilution by flow cytometry. Values are given as % proliferation relative to control used as 100%. The summarized data are shown as mean ± SD from 5 donors ** p < 0.01, *** p < 0.005, compared to control non-treated PHA-PBMC.
Figure 3
Figure 3
Representative histograms of PBMC proliferation. (A) control PHA-stimulated PBMC. (B) PEx = 40 μg/mL (C) PG = 40 μg/mL (D) PN = 40 μg/mL (E) EA = 5 μg/mL (F) EA = 10 μg/mL. (PI-proliferation index, DI-division index, div-percentage of division).
Figure 4
Figure 4
The effects of pomegranate peel extract (PEx), punicalagin (PG), punicalin (PN), ellagic acid (EA), and combinations of these ellagitannin components (C1 and C2) on pro-inflammatory cytokine production in PHA-stimulated PBMC cultures. PBMC cultures were treated with different concentrations of the compounds as indicated or with two different combinations of PG, PN and EA as indicated in Materials and methods. * p < 0.05, ** p < 0.01, *** p < 0.005, compared to control non-treated PBMC. # p < 0.05, ## p < 0.01, ### p < 0.005, compared to indicated individual concentrations of ellagitannins.
Figure 5
Figure 5
The effects of pomegranate peel extract (PEx), punicalagin (PG), punicalin (PN), ellagic acid (EA) and combinations of these ellagitannin components (C1 and C2) on T-cell cytokine production in PHA-stimulated PBMC cultures. PBMC cultures were treated with different concentrations of the compounds or with two different combinations of PG, PN and EA as indicated in Materials and methods. * p < 0.05, ** p < 0.01, *** p < 0.005, compared to control non-treated PBMC. # p < 0.05, ## p < 0.01, ### p < 0.005, compared to indicated concentrations of individual ellagitannins.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Baradaran Rahimi V., Ghadiri M., Ramezani M., Askari V.R. Antiinflammatory and anti-cancer activities of pomegranate and its constituent, ellagic acid: Evidence from cellular, animal, and clinical studies. Phytother. Res. 2020;34:685–720. doi: 10.1002/ptr.6565. - DOI - PubMed
    1. Ismail T., Sestili P., Akhtar S. Pomegranate peel and fruit extracts: A review of potential anti-inflammatory and anti-infective effects. J. Ethnopharmacol. 2012;143:397–405. doi: 10.1016/j.jep.2012.07.004. - DOI - PubMed
    1. Omer H.A.A., Abdel-Magid S.S., Awadalla I.M. Nutritional and chemical evaluation of dried pomegranate (Punica granatum L.) peels and studying the impact of level of inclusion in ration formulation on productive performance of growing Ossimi lambs. Bull. Natl. Res. Cent. 2019;43:182. doi: 10.1186/s42269-019-0245-0. - DOI
    1. Venusova E., Kolesarova A., Horky P., Slama P. Physiological and Immune Functions of Punicalagin. Nutrients. 2021;13:2150. doi: 10.3390/nu13072150. - DOI - PMC - PubMed
    1. Ismail T., Akhtar S., Sestili P., Riaz M., Ismail A., Labbe R.G. Antioxidant, Antimicrobial and Urease Inhibitory Activities of Phenolics-Rich Pomegranate Peel Hydro-Alcoholic Extracts. J. Food Biochem. 2016;40:550–558. doi: 10.1111/jfbc.12250. - DOI

LinkOut - more resources