Evaluation of Echinacea purpurea Extracts as Immunostimulants: Impact on Macrophage Activation

Abstract

Echinacea purpurea has been traditionally used to strengthen the immune system. Therefore, herein, we investigated the potential of E. purpurea aqueous extracts (AEs) obtained from flowers (F), leaves (L), or roots (R) as an immune booster in human primary monocyte‐derived macrophages (hMDMs). Additionally, to identify the main class of compounds (phenolic/carboxylic acids vs. alkylamides) responsible for the bioactivity, the three AEs were fractioned by semi-preparative high-performance liquid chromatography (HPLC). The AEs and the isolated phenolic/carboxylic acidic fractions were not cytotoxic for hMDMs for all tested concentrations, as confirmed by the metabolic activity and DNA content assays. Moreover, AE drastically induced the production of the interleukin (IL)-6 and tumor necrosis factor (TNF)-α, with a minimal effect on IL-1β and prostaglandin E2 (PGE2), supporting their potential for macrophage activation. Interestingly, in the presence of the phenolic/carboxylic acidic fractions, this efficacy considerably decreased, suggesting a complementary effect between compounds. AE also triggered the phosphorylation of the extracellular signal-regulated kinase (ERK) 1/2 and p38 signaling pathways and upregulated the cyclooxygenase (COX)-2 expression in hMDMs. Overall, AE-F was demonstrated to be the most powerful immunostimulant extract that can be related to their higher number in identified bioactive compounds compared to AE-L and AE-R. These results highlight the efficiency of E. purpurea AE to enhance the function of a key cell type of the immune system and their potential as immunostimulant formulations for patients with a compromised immune system due to certain diseases (e.g., acquired immunodeficiencies) and treatments (e.g., chemotherapy).

Keywords

Echinacea purpurea - Asteraceae - phenolic/carboxylic acids - inflammation - signaling pathways - human primary macrophages

Publikationsverlauf

Eingereicht: 12. Juni 2024

Angenommen nach Revision: 01. Oktober 2024

Artikel online veröffentlicht:
17. Oktober 2024

• References

• 1 WHO. WHO Monographs on Selected Medicinal Plants – Volume 1. Geneva: World Health Organization; 1999
• 2 Fu A, Wang Y, Wu Y, Chen H, Zheng S, Li Y, Xu X, Li W. Echinacea purpurea extract polarizes M1 macrophages in murine bone marrow-derived macrophages through the activation of JNK. J Cell Biochem 2017; 118: 2664-2671
• 3 Li Y, Wang Y, Wu Y, Wang B, Chen X, Xu X, Chen H, Li W, Xu X. Echinacea pupurea extracts promote murine dendritic cell maturation by activation of JNK, p 38 MAPK and NF-κB pathways. Dev Comp Immunol 2017; 73: 21-26
• 4 See DM, Broumand N, Sahl L, Tilles JG. In vitro effects of Echinacea and Ginseng on natural killer and antibody-dependent cell cytotoxicity in healthy subjects and chronic fatigue syndrome or acquired immunodeficiency syndrome patients. Immunopharmacology 1997; 35: 229-235
• 5 Stimpel M, Proksch A, Wagner H, Lohmann-Matthes ML. Macrophage activation and induction of macrophage cytotoxicity by purified polysaccharide fractions from the plant Echinacea purpurea . Infect Immun 1984; 46: 845-849
• 6 Yang G, Li K, Liu C, Peng P, Bai M, Sun J, Li Q, Yang Z, Yang Y, Wu H. A comparison of the immunostimulatory effects of polysaccharides from tetraploid and diploid Echinacea purpurea . Biomed Res Int 2018; 2018: 1-12
• 7 Ramasahayam S, Baraka HN, Abdel Bar FM, Abuasal BS, Widrlechner MP, Sayed KA, Meyer SA. Effects of chemically characterized fractions from aerial parts of Echinacea purpurea and E. angustifolia on myelopoiesis in rats. Planta Med 2011; 77: 1883-1889
• 8 Hwang SA, Dasgupta A, Actor JK. Cytokine production by non-adherent mouse splenocyte cultures to Echinacea extracts. Clin Chim Acta 2004; 343: 161-166
• 9 Sullivan AM, Laba JG, Moore JA, Lee TDG. Echinacea-induced macrophage activation. Immunopharmacol Immunotoxicol 2008; 30: 553-574
• 10 Randolph RK, Gellenbeck K, Stonebrook K, Brovelli E, Qian Y, Bankaitis-Davis D, Cheronis J. Regulation of human immune gene expression as influenced by a commercial blended Echinacea product: Preliminary studies. Exp Biol Med 2003; 228: 1051-1056
• 11 Benson JM, Pokorny AJ, Rhule A, Wenner CA, Kandhi V, Cech NB, Shepherd DM. Echinacea purpurea extracts modulate murine dendritic cell fate and function. Food Chem Toxicol 2010; 48: 1170-1177
• 12 Wang CY, Staniforth V, Chiao MT, Hou CC, Wu HM, Yeh KC, Chen CH, Hwang PI, Wen TN, Shyur LF, Yang NS. Genomics and proteomics of immune modulatory effects of a butanol fraction of Echinacea purpurea in human dendritic cells. BMC Genomics 2008; 9: 1-20
• 13 Zwickey H, Brush J, Iacullo CM, Connelly E, Gregory WL, Soumyanath A, Buresh R. The effect of Echinacea purpurea, Astragalus membranaceus and Glycyrrhiza glabra on CD25 expression in humans: A pilot study. Phyther Res 2007; 21: 1109-1112
• 14 Brush J, Mendenhall E, Guggenheim A, Chan T, Connelly E, Soumyanath A, Buresh R, Barrett R, Zwickey H. The effect of Echinacea purpurea, Astragalus membranaceus and Glycyrrhiza glabra on CD69 expression and immune cell activation in humans. Phytother Res 2006; 20: 687-695
• 15 Wang CY, Chiao MT, Yen PJ, Huang WC, Hou CC, Chien SC, Yeh KC, Yang WC, Shyur LF, Yang NS. Modulatory effects of Echinacea purpurea extracts on human dendritic cells: A cell- and gene-based study. Genomics 2006; 88: 801-808
• 16 Gan XH, Zhang L, Heber D, Bonavida B. Mechanism of activation of human peripheral blood NK cells at the single cell level by Echinacea water soluble extracts: Recruitment of lymphocyte–target conjugates and killer cells and activation of programming for lysis. Int Immunopharmacol 2003; 3: 811-824
• 17 Currier NL, Miller SC. Natural killer cells from aging mice treated with extracts from Echinacea purpurea are quantitatively and functionally rejuvenated. Exp Gerontol 2000; 35: 627-639
• 18 Rininger JA, Kickner S, Chigurupati P, McLean A, Franck Z. Immunopharmacological activity of Echinacea preparations following simulated digestion on murine macrophages and human peripheral blood mononuclear cells. J Leukoc Biol 2000; 68: 503-510
• 19 Yamada K, Hung P, Park TK, Park PJ, Lim BO. A comparison of the immunostimulatory effects of the medicinal herbs Echinacea, Ashwagandha and Brahmi. J Ethnopharmacol 2011; 137: 231-235
• 20 Gertsch J, Schoop R, Kuenzle U, Suter A. Echinacea alkylamides modulate TNF-alpha gene expression via cannabinoid receptor CB2 and multiple signal transduction pathways. FEBS Lett 2004; 577: 563-569
• 21 Goel V, Chang C, Slama JV, Barton R, Bauer R, Gahler R, Basu TK. Alkylamides of Echinacea purpurea stimulate alveolar macrophage function in normal rats. Int Immunopharmacol 2002; 2: 381-387
• 22 Goel V, Chang C, Slama JV, Barton R, Bauer R, Gahler R, Basu TK. Echinacea stimulates macrophage function in the lung and spleen of normal rats. J Nutr Biochem 2002; 13: 487-492
• 23 Burger RA, Torres AR, Warren RP, Caldwell VD, Hughes BG. Echinacea-induced cytokine production by human macrophages. Int J Immunopharmacol 1997; 19: 371-379
• 24 Freier DO, Wright K, Klein K, Voll D, Dabiri K, Cosulich K, George R. Enhancement of the humoral immune response by Echinacea purpurea in female Swiss mice. Immunopharmacol Immunotoxicol 2003; 25: 551-560
• 25 Sun LZ, Currier NL, Miller SC. The American coneflower: A prophylactic role involving nonspecific immunity. J Altern Complement Med 1999; 5: 437-446
• 26 Yin SY, Wang WH, Wang BX, Aravindaram K, Hwang PI, Wu HM, Yang NS. Stimulatory effect of Echinacea purpurea extract on the trafficking activity of mouse dendritic cells: Revealed by genomic and proteomic analyses. BMC Genomics 2010; 11: 1-18
• 27 Fonseca FN, Papanicolaou G, Lin H, Lau CBS, Kennelly EJ, Cassileth BR, Cunningham-Rundles S. Echinacea purpurea (L.) Moench modulates human T-cell cytokine response. Int Immunopharmacol 2014; 19: 94-102
• 28 Goel V, Lovlin R, Chang C, Slama JV, Barton R, Gahler R, Bauer R, Goonewardene L, Basu TK. A proprietary extract from the echinacea plant (Echinacea purpurea) enhances systemic immune response during a common cold. Phytother Res 2005; 19: 689-694
• 29 Hirayama D, Iida T, Nakase H. The phagocytic function of macrophage-enforcing innate immunity and tissue homeostasis. Int J Mol Sci 2017; 19: 1-14
• 30 Turner MD, Nedjai B, Hurst T, Pennington DJ. Cytokines and chemokines: At the crossroads of cell signalling and inflammatory disease. Biochim Biophys Acta 2014; 1843: 2563-2582
• 31 Dorrington MG, Fraser IDC. NF-κB signaling in macrophages: Dynamics, crosstalk, and signal integration. Front Immunol 2019; 10: 1-12
• 32 Liu T, Zhang L, Joo D, Sun SC. NF-κB signaling in inflammation. Signal Transduct Target Ther 2017; 2: 1-9
• 33 Vieira SF, Gonçalves VMF, Llaguno CP, Macías F, Tiritan ME, Reis RL, Ferreira H, Neves NM. On the bioactivity of Echinacea purpurea extracts to modulate the production of inflammatory mediators. Int J Mol Sci 2022; 23: 1-25
• 34 Medzhitov R. Origin and physiological roles of inflammation. Nature 2008; 454: 428-435
• 35 Nicholson LB. The immune system. Essays Biochem 2016; 60: 275-301
• 36 Newman DJ, Cragg GM. Natural products as sources of new drugs over the nearly four decades from 01/1981 to 09/2019. J Nat Prod 2020; 83: 770-803
• 37 Matthias A, Banbury L, Bone KM, Leach DN, Lehmann RP. Echinacea alkylamides modulate induced immune responses in T-cells. Fitoterapia 2008; 79: 53-58
• 38 Matthias A, Blanchfield JT, Penman KG, Toth I, Lang CS, De Voss JJ, Lehmann RP. Permeability studies of alkylamides and caffeic acid conjugates from echinacea using a Caco-2 cell monolayer model. J Clin Pharm Ther 2004; 29: 7-13
• 39 Matthias A, Penman K, Matovic N, Bone K, De Voss J, Lehmann R. Bioavailability of Echinacea constituents: Caco-2 monolayers and pharmacokinetics of the alkylamides and caffeic acid conjugates. Molecules 2005; 10: 1242-1251
• 40 Matthias A, Addison RS, Penman KG, Dickinson RG, Bone KM, Lehmann RP. Echinacea alkamide disposition and pharmacokinetics in humans after tablet ingestion. Life Sci 2005; 77: 2018-2029
• 41 Dietz B, Heilmann J, Bauer R. Absorption of dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamides after oral application of Echinacea purpurea tincture. Planta Med 2001; 67: 863-864
• 42 Vieira SF, Gonçalves SM, Gonçalves VMF, Llaguno CP, Macías F, Tiritan ME, Cunha C, Carvalho A, Reis RL, Ferreira H, Neves NM. Echinacea purpurea fractions represent promising plant-based anti-inflammatory formulations. Antioxidants 2023; 12: 1-25
• 43 Gonçalves SM, Duarte-Oliveira C, Campos CF, Aimanianda V, Ter Horst R, Leite L, Mercier T, Pereira P, Fernández-García M, Antunes D, Rodrigues CS, Barbosa-Matos C, Gaifem J, Mesquita I, Marques A, Osório NS, Torrado E, Rodrigues F, Costa A, Joosten LAB, Lagrou K, Maertens J, Lacerda JF, Campos jr. A, Brown GD, Brakhage AA, Barbas C, Silvestre R, van de Veerdonk FL, Chamilos G, Netea MG, Latgé JP, Cunha C, Carvalho A. Phagosomal removal of fungal melanin reprograms macrophage metabolism to promote antifungal immunity. Nat Commun 2020; 11: 1-15
• 44 Vieira SF, Ferreira H, Neves NM. Antioxidant and anti-inflammatory activities of cytocompatible Salvia officinalis extracts: A comparison between traditional and soxhlet extraction. Antioxidants 2020; 9: 1-25