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Year 2025, Volume: 29 Issue: 3, 1028 - 1037, 04.06.2025
https://doi.org/10.12991/jrespharm.1694190

Abstract

References

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  • [2] Medina KL. Overview of the immune system. In: Handbook of clinical neurology. Elsevier; 2016. p. 61–76. https://doi.org/10.1016/B978-0-444-63432-0.00004-9
  • [3] Brodin P, Davis MM. Human immune system variation. Nat Rev Immunol. 2017;17(1):21–29. https://doi.org/10.1038/nri.2016.125.
  • [4] Feng X, Xu W, Li Z, Song W, Ding J, Chen X. Immunomodulatory Nanosystems. Adv Sci. 2019;6(17). https://doi.org/10.1002/advs.201900101.
  • [5] Kumar P, Rai S, Verma SK, Prakash PS, Chitara D. Classification, mode of action and uses of various immunomodulators. In: Immunomodulators and human health. Springer; 2022. p. 3–38.
  • [6] Singh N, Tailang M, Mehta SC. A revıew on herbal plants as ımmunomodulators. Int J Pharm Sci Res. 2016;7(9):3602–3610. http://dx.doi.org/10.13040/IJPSR.0975-8232.7 (9).3602-3610.
  • [7] Zebeaman M, Tadesse MG, Bachheti RK, Bachheti A, Gebeyhu R, Chaubey KK. plants and plant‐derived molecules as natural ımmunomodulators. Biomed Res Int. 2023;2023(1):7711297. https://doi.org/10.1155/2023/7711297
  • [8] Atanassova M, Martorell M, Sharopov F, Atanassov L, Kumar P, Sharifi-Rad J, Tejada-Gavela S, Iriti M, Pezzani R, Varoni EM. Cocoa as immunomodulatory agent: an update. Nat Prod Res. 2024;38(23):4196-4207. https://doi.org/10.1080/14786419.2023.2272025.
  • [9] Ben Lagha A, Maquera Huacho P, Grenier D. A cocoa (Theobroma cacao L.) extract impairs the growth, virulence properties, and inflammatory potential of Fusobacterium nucleatum and improves oral epithelial barrier function. PLoS One. 2021;16(5):e0252029. https://doi.org/10.1371/journal.pone.0252029.
  • [10] Sorrenti V, Ali S, Mancin L, Davinelli S, Paoli A, Scapagnini G. Cocoa polyphenols and gut microbiota interplay: Bioavailability, prebiotic effect, and impact on human health. Nutrients. 2020;12(7):1908. https://doi.org/10.3390/nu12071908.
  • [11] Ramos-Escudero F, Casimiro-Gonzales S, Fernández-Prior Á, Chávez KC, Gómez-Mendoza J, de la Fuente-Carmelino L, Munoz AM. Colour, fatty acids, bioactive compounds, and total antioxidant capacity in commercial cocoa beans (Theobroma cacao L.). LWT. 2021;147:111629. https://doi.org/10.1016/j.lwt.2021.111629.
  • [12] Lee KW, Kim YJ, Lee HJ, Lee CY. Cocoa has more phenolic phytochemicals and a higher antioxidant capacity than teas and red wine. J Agric Food Chem. 2003;51(25):7292–7295. https://doi.org/10.1021/jf0344385.
  • [13] Osman H, Nasarudin R, Lee SL. Extracts of cocoa (Theobroma cacao L.) leaves and their antioxidation potential. Food Chem. 2004;86(1):41–46. https://doi.org/10.1016/j.foodchem.2003.08.026.
  • [14] Othman A, Ismail A, Abdul Ghani N, Adenan I. Antioxidant capacity and phenolic content of cocoa beans. Food Chem. 2007;100(4):1523–1530. https://doi.org/10.1016/j.foodchem.2005.12.021.
  • [15] Zeng H, Locatelli M, Bardelli C, Amoruso A, Coisson JD, Travaglia F, Arlorio M, Brunelleschi S. Anti-inflammatory properties of clovamide and Theobroma cacao phenolic extracts in human monocytes: evaluation of respiratory burst, cytokine release, NF-κB activation, and PPARγ modulation. J Agric Food Chem. 2011;59(10):5342-5350. https://doi.org/10.1021/jf2005386.
  • [16] Okwori AEJ, Olayanju OA, Wali CC, Kolawole TA, Oluwatayo BO. Possible antioxidant and haematinic properties of the stem bark of Theobroma cacao L. in Wistar Rats. Int J Biochem Res Rev. 2021;18–26. https://doi.org/10.9734/ijbcrr/2021/v30i330256.
  • [17] Vargas-Munévar L, Borja-Fajardo J, Sandoval-Aldana A, García WQ, Moreno EM, Henriquez JC, Stashenko E, García LT, García-Beltrán O. Microencapsulation of Theobroma cacao L polyphenols: A high-value approach with in vitro anti-Trypanosoma cruzi, immunomodulatory and antioxidant activities. Biomed Pharmacother. 2024;173:116307. https://doi.org/10.1016/j.biopha.2024.116307.
  • [18] Lee HW, Choi IW, Ha SK. Immunostimulatory activities of theobromine on macrophages via the activation of MAPK and NF-κB signaling pathways. Curr Issues Mol Biol. 2022;44(9):4216–4228. https://doi.org/10.3390/cimb44090289.
  • [19] Rahayu YC, Setiawatie EM, Rahayu RP, Sakinah NN, Kusumawardani B, Gunadi A, Ryandhana PH. Effects of cocoa pod husk extract (Theobroma cacao L.) on alveolar bone in experimental periodontitis rats. Trends Sci. 2023;20(6):6535. https://orcid.org/0000-0002-9915-0468.
  • [20] Rahayu YC, Setiawatie EM, Rahayu RP, Cholid Z, Rachmawati D, Sulistyani E, Hamidah S. Wound healing potential of cocoa pod husk (Theobroma cacao L.) extract gel on tooth extraction socket in rats. Mal J Med Health Sci 2024; 20(SUPP5): 11-16.
  • [21] Aprila Fajrin F, Holidah D, Nurhidayah H, Suci Wulansari P, Pudji Restanto D, Azkiyah L, Witono Y, Satia Nugraha A. The ethanol extract of cocoa pod husk minimizes hyperalgesia and blood glucose levels in diabetic neuropathy model through transient receptor protein vanilloid (TRPV)-1. Saudi Pharm J. 2024;32(6):102097. https://doi.org/10.1016/j.jsps.2024.102097.
  • [22] Jacob S, Nair AB, Shah J. Emerging role of nanosuspensions in drug delivery systems. Biomater Res. 2020;24(1):3. https://doi.org/10.1186/s40824-020-0184-8.
  • [23] Dewangan HK. The emerging role of nanosuspensions for drug delivery and stability. Curr Nanomed. 2021;11(4):213–223. https://doi.org/10.2174/2468187312666211222123307.
  • [24] Chinthaginjala H, Abdul H, Reddy APG, Kodi K, Manchikanti SP, Pasam D. Nanosuspension as promising and potential drug delivery: A review. Int J Life Sci Pharm Res. 2020;11(1):P59-66. http://dx.doi.org/10.22376/ijpbs/lpr.2021.11.1.P59-66.
  • [25] Wagner H. Immunostimulants from higher plants (recent advances). Biologically active natural products. K Hostettmann and PJ Lea (Editors), 1987.
  • [26] Aulia ISI, Wijayanti T, Saptarini O. Immunomodulatory activity of ethanol extract and Kabau (Archidendron bubalinum) leaf fractions on the macrophage phagocytosis ındex in male white rats (Rattus Novergicus). Jurnal Multidisiplin Madani. 2024;4(6):830–842. https://doi.org/10.55927/mudima.v4i6.9588.
  • [27] Koyama Y, Tomoda Y, Kato M, Ashihara H. Metabolism of purine bases, nucleosides and alkaloids in theobromine-forming Theobroma cacao leaves. Plant Physiol Biochem. 2003;41(11–12):977–984. https://doi.org/10.1016/j.plaphy.2003.07.002.
  • [28] Abulude FO, Ogunkoya MO, Adenibuyan GB, Arifalo KM, Akinusotu A, Samuel A, Adamu A, Kenni AM, Bello LJ. Phytochemical assessment of the extracts of stem (Bark) and leaves of Theobroma cocoa materials: Experimental procedure and its comparison to literature. ASEAN J Sci Eng Mater. 2022;1(2):85–92.
  • [29] Kementerian Kesehatan RI. Farmakope Herbal Indonesia Edisi II . Jakarta: Kementerian Kesehatan RI; 2017.
  • [30] Gao B. Basic liver immunology. Cell Mol Immunol. 2016;13(3):265–266. https://doi.org/10.1038/cmi.2016.09.
  • [31] Bronte V, Pittet MJ. The spleen in local and systemic regulation of ımmunity. Immunity. 2013; 39(5):806–818. https://doi.org/10.1016/j.immuni.2013.10.010.
  • [32] Raphael I, Nalawade S, Eagar TN, Forsthuber TG. T cell subsets and their signature cytokines in autoimmune and inflammatory diseases. Cytokine. 2015;74(1):5–17. https://doi.org/10.1016/j.cyto.2014.09.011.
  • [33] Kato H, Perl A. Double-Edged Sword: Interleukin-2 promotes T regulatory cell differentiation but also expands Interleukin-13- and Interferon-γ-Producing CD8+ T Cells via STAT6-GATA-3 axis in Systemic Lupus Erythematosus. Front Immunol. 2021;12:635531. https://doi.org/10.3389/fimmu.2021.635531.
  • [34] Sim GC, Radvanyi L. The IL-2 cytokine family in cancer immunotherapy. Cytokine Growth Factor Rev. 2014;25(4):377–390. https://doi.org/10.1016/j.cytogfr.2014.07.018.

Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice

Year 2025, Volume: 29 Issue: 3, 1028 - 1037, 04.06.2025
https://doi.org/10.12991/jrespharm.1694190

Abstract

This study intends to examine the immunomodulatory effects of cocoa (Theobroma cacao L.) leaves in vivo in mice using two types of comparison between the crude extract form and the nanosuspension form. Cocoa leaf extract is made from young leaves (YL) and old leaves (OL), where the extraction of active compounds from cocoa leaves was carried out by maceration using 96% ethanol. YL and OL in crude extract and nanosuspensions were tested for their immunomodulatory effects in vivo in BALB/c mice through tests against nonspecific and specific immune responses. Tests for nonspecific immune responses include carbon clearance tests, organ index determination, and testing against specific immune responses, including measurement of antibody titers as humoral immune responses and cytokine levels as a cellular immune response. Results showed that the three doses of OL and YL extract (i.e., 50,100,200 mg/kg BW), both in crude form extract and nanosuspensions, had immunosuppressive effects characterized by phagocytic index <1. YL and OL have the same immunosuppressive effect. Extract in nanosuspensions show a more substantial immunosuppressive effect than crude extract. In testing against specific immune responses, all three doses of extract in both crude form extract and nanosuspension inhibited antibody, IFN-γ, and IL-2 formation. Based on those results, it can be concluded that all three doses of YL/OL in both crude and nanosuspension forms are immunosuppressive.

References

  • [1] Nicholson LB. The immune system. Essays Biochem. 2016;60(3):275–301. https://doi.org/10.1042/EBC20160017.
  • [2] Medina KL. Overview of the immune system. In: Handbook of clinical neurology. Elsevier; 2016. p. 61–76. https://doi.org/10.1016/B978-0-444-63432-0.00004-9
  • [3] Brodin P, Davis MM. Human immune system variation. Nat Rev Immunol. 2017;17(1):21–29. https://doi.org/10.1038/nri.2016.125.
  • [4] Feng X, Xu W, Li Z, Song W, Ding J, Chen X. Immunomodulatory Nanosystems. Adv Sci. 2019;6(17). https://doi.org/10.1002/advs.201900101.
  • [5] Kumar P, Rai S, Verma SK, Prakash PS, Chitara D. Classification, mode of action and uses of various immunomodulators. In: Immunomodulators and human health. Springer; 2022. p. 3–38.
  • [6] Singh N, Tailang M, Mehta SC. A revıew on herbal plants as ımmunomodulators. Int J Pharm Sci Res. 2016;7(9):3602–3610. http://dx.doi.org/10.13040/IJPSR.0975-8232.7 (9).3602-3610.
  • [7] Zebeaman M, Tadesse MG, Bachheti RK, Bachheti A, Gebeyhu R, Chaubey KK. plants and plant‐derived molecules as natural ımmunomodulators. Biomed Res Int. 2023;2023(1):7711297. https://doi.org/10.1155/2023/7711297
  • [8] Atanassova M, Martorell M, Sharopov F, Atanassov L, Kumar P, Sharifi-Rad J, Tejada-Gavela S, Iriti M, Pezzani R, Varoni EM. Cocoa as immunomodulatory agent: an update. Nat Prod Res. 2024;38(23):4196-4207. https://doi.org/10.1080/14786419.2023.2272025.
  • [9] Ben Lagha A, Maquera Huacho P, Grenier D. A cocoa (Theobroma cacao L.) extract impairs the growth, virulence properties, and inflammatory potential of Fusobacterium nucleatum and improves oral epithelial barrier function. PLoS One. 2021;16(5):e0252029. https://doi.org/10.1371/journal.pone.0252029.
  • [10] Sorrenti V, Ali S, Mancin L, Davinelli S, Paoli A, Scapagnini G. Cocoa polyphenols and gut microbiota interplay: Bioavailability, prebiotic effect, and impact on human health. Nutrients. 2020;12(7):1908. https://doi.org/10.3390/nu12071908.
  • [11] Ramos-Escudero F, Casimiro-Gonzales S, Fernández-Prior Á, Chávez KC, Gómez-Mendoza J, de la Fuente-Carmelino L, Munoz AM. Colour, fatty acids, bioactive compounds, and total antioxidant capacity in commercial cocoa beans (Theobroma cacao L.). LWT. 2021;147:111629. https://doi.org/10.1016/j.lwt.2021.111629.
  • [12] Lee KW, Kim YJ, Lee HJ, Lee CY. Cocoa has more phenolic phytochemicals and a higher antioxidant capacity than teas and red wine. J Agric Food Chem. 2003;51(25):7292–7295. https://doi.org/10.1021/jf0344385.
  • [13] Osman H, Nasarudin R, Lee SL. Extracts of cocoa (Theobroma cacao L.) leaves and their antioxidation potential. Food Chem. 2004;86(1):41–46. https://doi.org/10.1016/j.foodchem.2003.08.026.
  • [14] Othman A, Ismail A, Abdul Ghani N, Adenan I. Antioxidant capacity and phenolic content of cocoa beans. Food Chem. 2007;100(4):1523–1530. https://doi.org/10.1016/j.foodchem.2005.12.021.
  • [15] Zeng H, Locatelli M, Bardelli C, Amoruso A, Coisson JD, Travaglia F, Arlorio M, Brunelleschi S. Anti-inflammatory properties of clovamide and Theobroma cacao phenolic extracts in human monocytes: evaluation of respiratory burst, cytokine release, NF-κB activation, and PPARγ modulation. J Agric Food Chem. 2011;59(10):5342-5350. https://doi.org/10.1021/jf2005386.
  • [16] Okwori AEJ, Olayanju OA, Wali CC, Kolawole TA, Oluwatayo BO. Possible antioxidant and haematinic properties of the stem bark of Theobroma cacao L. in Wistar Rats. Int J Biochem Res Rev. 2021;18–26. https://doi.org/10.9734/ijbcrr/2021/v30i330256.
  • [17] Vargas-Munévar L, Borja-Fajardo J, Sandoval-Aldana A, García WQ, Moreno EM, Henriquez JC, Stashenko E, García LT, García-Beltrán O. Microencapsulation of Theobroma cacao L polyphenols: A high-value approach with in vitro anti-Trypanosoma cruzi, immunomodulatory and antioxidant activities. Biomed Pharmacother. 2024;173:116307. https://doi.org/10.1016/j.biopha.2024.116307.
  • [18] Lee HW, Choi IW, Ha SK. Immunostimulatory activities of theobromine on macrophages via the activation of MAPK and NF-κB signaling pathways. Curr Issues Mol Biol. 2022;44(9):4216–4228. https://doi.org/10.3390/cimb44090289.
  • [19] Rahayu YC, Setiawatie EM, Rahayu RP, Sakinah NN, Kusumawardani B, Gunadi A, Ryandhana PH. Effects of cocoa pod husk extract (Theobroma cacao L.) on alveolar bone in experimental periodontitis rats. Trends Sci. 2023;20(6):6535. https://orcid.org/0000-0002-9915-0468.
  • [20] Rahayu YC, Setiawatie EM, Rahayu RP, Cholid Z, Rachmawati D, Sulistyani E, Hamidah S. Wound healing potential of cocoa pod husk (Theobroma cacao L.) extract gel on tooth extraction socket in rats. Mal J Med Health Sci 2024; 20(SUPP5): 11-16.
  • [21] Aprila Fajrin F, Holidah D, Nurhidayah H, Suci Wulansari P, Pudji Restanto D, Azkiyah L, Witono Y, Satia Nugraha A. The ethanol extract of cocoa pod husk minimizes hyperalgesia and blood glucose levels in diabetic neuropathy model through transient receptor protein vanilloid (TRPV)-1. Saudi Pharm J. 2024;32(6):102097. https://doi.org/10.1016/j.jsps.2024.102097.
  • [22] Jacob S, Nair AB, Shah J. Emerging role of nanosuspensions in drug delivery systems. Biomater Res. 2020;24(1):3. https://doi.org/10.1186/s40824-020-0184-8.
  • [23] Dewangan HK. The emerging role of nanosuspensions for drug delivery and stability. Curr Nanomed. 2021;11(4):213–223. https://doi.org/10.2174/2468187312666211222123307.
  • [24] Chinthaginjala H, Abdul H, Reddy APG, Kodi K, Manchikanti SP, Pasam D. Nanosuspension as promising and potential drug delivery: A review. Int J Life Sci Pharm Res. 2020;11(1):P59-66. http://dx.doi.org/10.22376/ijpbs/lpr.2021.11.1.P59-66.
  • [25] Wagner H. Immunostimulants from higher plants (recent advances). Biologically active natural products. K Hostettmann and PJ Lea (Editors), 1987.
  • [26] Aulia ISI, Wijayanti T, Saptarini O. Immunomodulatory activity of ethanol extract and Kabau (Archidendron bubalinum) leaf fractions on the macrophage phagocytosis ındex in male white rats (Rattus Novergicus). Jurnal Multidisiplin Madani. 2024;4(6):830–842. https://doi.org/10.55927/mudima.v4i6.9588.
  • [27] Koyama Y, Tomoda Y, Kato M, Ashihara H. Metabolism of purine bases, nucleosides and alkaloids in theobromine-forming Theobroma cacao leaves. Plant Physiol Biochem. 2003;41(11–12):977–984. https://doi.org/10.1016/j.plaphy.2003.07.002.
  • [28] Abulude FO, Ogunkoya MO, Adenibuyan GB, Arifalo KM, Akinusotu A, Samuel A, Adamu A, Kenni AM, Bello LJ. Phytochemical assessment of the extracts of stem (Bark) and leaves of Theobroma cocoa materials: Experimental procedure and its comparison to literature. ASEAN J Sci Eng Mater. 2022;1(2):85–92.
  • [29] Kementerian Kesehatan RI. Farmakope Herbal Indonesia Edisi II . Jakarta: Kementerian Kesehatan RI; 2017.
  • [30] Gao B. Basic liver immunology. Cell Mol Immunol. 2016;13(3):265–266. https://doi.org/10.1038/cmi.2016.09.
  • [31] Bronte V, Pittet MJ. The spleen in local and systemic regulation of ımmunity. Immunity. 2013; 39(5):806–818. https://doi.org/10.1016/j.immuni.2013.10.010.
  • [32] Raphael I, Nalawade S, Eagar TN, Forsthuber TG. T cell subsets and their signature cytokines in autoimmune and inflammatory diseases. Cytokine. 2015;74(1):5–17. https://doi.org/10.1016/j.cyto.2014.09.011.
  • [33] Kato H, Perl A. Double-Edged Sword: Interleukin-2 promotes T regulatory cell differentiation but also expands Interleukin-13- and Interferon-γ-Producing CD8+ T Cells via STAT6-GATA-3 axis in Systemic Lupus Erythematosus. Front Immunol. 2021;12:635531. https://doi.org/10.3389/fimmu.2021.635531.
  • [34] Sim GC, Radvanyi L. The IL-2 cytokine family in cancer immunotherapy. Cytokine Growth Factor Rev. 2014;25(4):377–390. https://doi.org/10.1016/j.cytogfr.2014.07.018.
There are 34 citations in total.

Details

Primary Language English
Subjects Pharmacology and Pharmaceutical Sciences (Other)
Journal Section Articles
Authors

Vita Olivia Siregar

Rachmat Mauludın

Maria Immaculata Iwo

Publication Date June 4, 2025
Submission Date May 3, 2024
Acceptance Date August 1, 2024
Published in Issue Year 2025 Volume: 29 Issue: 3

Cite

APA Siregar, V. O., Mauludın, R., & Iwo, M. I. (2025). Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice. Journal of Research in Pharmacy, 29(3), 1028-1037. https://doi.org/10.12991/jrespharm.1694190
AMA Siregar VO, Mauludın R, Iwo MI. Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice. J. Res. Pharm. June 2025;29(3):1028-1037. doi:10.12991/jrespharm.1694190
Chicago Siregar, Vita Olivia, Rachmat Mauludın, and Maria Immaculata Iwo. “Immunomodulator Activity of Nanosuspension of Cocoa (Theobroma Cacao L.) Leaf Extract in Balb C Mice”. Journal of Research in Pharmacy 29, no. 3 (June 2025): 1028-37. https://doi.org/10.12991/jrespharm.1694190.
EndNote Siregar VO, Mauludın R, Iwo MI (June 1, 2025) Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice. Journal of Research in Pharmacy 29 3 1028–1037.
IEEE V. O. Siregar, R. Mauludın, and M. I. Iwo, “Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice”, J. Res. Pharm., vol. 29, no. 3, pp. 1028–1037, 2025, doi: 10.12991/jrespharm.1694190.
ISNAD Siregar, Vita Olivia et al. “Immunomodulator Activity of Nanosuspension of Cocoa (Theobroma Cacao L.) Leaf Extract in Balb C Mice”. Journal of Research in Pharmacy 29/3 (June2025), 1028-1037. https://doi.org/10.12991/jrespharm.1694190.
JAMA Siregar VO, Mauludın R, Iwo MI. Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice. J. Res. Pharm. 2025;29:1028–1037.
MLA Siregar, Vita Olivia et al. “Immunomodulator Activity of Nanosuspension of Cocoa (Theobroma Cacao L.) Leaf Extract in Balb C Mice”. Journal of Research in Pharmacy, vol. 29, no. 3, 2025, pp. 1028-37, doi:10.12991/jrespharm.1694190.
Vancouver Siregar VO, Mauludın R, Iwo MI. Immunomodulator activity of nanosuspension of cocoa (Theobroma cacao L.) leaf extract in Balb/c mice. J. Res. Pharm. 2025;29(3):1028-37.