RSS-Feed abonnieren
DOI: 10.1055/a-0637-9936
Magnoflorine Enhances LPS-Activated Pro-Inflammatory Responses via MyD88-Dependent Pathways in U937 Macrophages
Publikationsverlauf
received 29. März 2018
revised 17. Mai 2018
accepted 28. Mai 2018
Publikationsdatum:
15. Juni 2018 (online)
![](https://www.thieme-connect.de/media/plantamedica/201817/lookinside/thumbnails/10-1055-a-0637-9936_pmc0260-1.jpg)
Abstract
Magnoflorine, a major bioactive metabolite isolated from Tinospora crispa, has been reported for its diverse biochemical and pharmacological properties. However, there is little report on its underlying mechanisms of action on immune responses, particularly on macrophage activation. In this study, we aimed to investigate the effects of magnoflorine, isolated from T. crispa on the pro-inflammatory mediators generation induced by LPS and the concomitant NF-κB, MAPKs, and PI3K-Akt signaling pathways in U937 macrophages. Differentiated U937 macrophages were treated with magnoflorine and the release of pro-inflammatory mediators was evaluated through ELISA, while the relative mRNA expression of the respective mediators was quantified through qRT-PCR. Correspondingly, western blotting was executed to observe the modulatory effects of magnoflorine on the expression of various markers related to NF-κB, MAPK and PI3K-Akt signaling activation in LPS-primed U937 macrophages. Magnoflorine significantly enhanced the upregulation of TNF-α, IL-1β, and PGE2 production as well as COX-2 protein expression. Successively, magnoflorine prompted the mRNA transcription level of these pro-inflammatory mediators. Magnoflorine enhanced the NF-κB activation by prompting p65, IκBα, and IKKα/β phosphorylation as well as IκBα degradation. Besides, magnoflorine treatments concentration-dependently augmented the phosphorylation of JNK, ERK, and p38 MAPKs as well as Akt. The immunoaugmenting effects were further confirmed by investigating the effects of magnoflorine on specific inhibitors, where the treatment with specific inhibitors of NF-κB, MAPKs, and PI3K-Akt proficiently blocked the magnoflorine-triggered TNF-α release and COX-2 expression. Magnoflorine furthermore enhanced the MyD88 and TLR4 upregulation. The results suggest that magnoflorine has high potential on augmenting immune responses.
Key words
magnoflorine - Tinospora crispa - Menispermaceae - immunomodulation - MyD88 - NF-κB - MAPKs - AktSupporting Information
- Supporting Information
1H NMR and 13C NMR spectra of MAG are available as Supporting Information.
-
References
- 1 Huang TT, Chong KY, Ojcius DM, Wu YH, Ko YF, Wu CY, Martel J, Lu CC, Lai HC, Young JD. Hirsutella sinensis mycelium suppresses interleukin-1β and interleukin-18 secretion by inhibiting both canonical and non-canonical inflammasomes. Sci Rep 2013; 3: 1374
- 2 Kim TH, Lee SJ, Rim HK, Shin JS, Jung JY, Heo JS, Kim JB, Lee MS, Lee KT. In vitro and in vivo immunostimulatory effects of hot water extracts from the leaves of Artemisia princeps Pampanini cv. Sajabal. J Ethnopharmacol 2013; 149: 254-262
- 3 Han EH, Choi JH, Hwang YP, Park HJ, Choi CY, Chung YC, Seo JK, Jeong HG. Immunostimulatory activity of aqueous extract isolated from Prunella vulgaris . Food Chem Toxicol 2009; 47: 62-69
- 4 Bauerfeld CP, Rastogi R, Pirockinaite G, Lee I, Hüttemann M, Monks B, Birnbaum MJ, Franchi L, Nuñez G, Samavati L. TLR4-mediated AKT activation is MyD88/TRIF dependent and critical for induction of oxidative phosphorylation and mitochondrial transcription factor A in murine macrophages. J Immunol 2012; 188: 2847-2857
- 5 Jang SE, Hyam SR, Jeong JJ, Han MJ, Kim DH. Penta-O-galloyl-β-D-glucose ameliorates inflammation by inhibiting MyD88/NF-κB and MyD88/MAPK signalling pathways. Br J Pharmacol 2013; 170: 1078-1091
- 6 Shin S, Kwon J, Lee S, Kong H, Lee S, Lee CK, Cho K, Ha NJ, Kim K. Immunostimulatory effects of Cordyceps militaris on macrophages through the enhanced production of cytokines via the activation of NF-κB. Immune Netw 2010; 10: 55-63
- 7 Patel MB, Mishra S. Hypoglycemic activity of alkaloidal fraction of Tinospora cordifolia . Phytomedicine 2011; 18: 1045-1052
- 8 Patel MB, Mishra SM. Magnoflorine from Tinospora cordifolia stem inhibits α-glucosidase and is antiglycemic in rats. J Funct Foods 2012; 4: 79-86
- 9 El Tahir KE. Pharmacological actions of magnoflorine and aristolochic acid-1 isolated from the seeds of Aristolochia bracteata . Int J Pharmacogn 1991; 29: 101-110
- 10 Rashid MA, Gustafson KR, Kashman Y, Cardellina JH, McMahon JB, Boyd MR. Anti-HIV alkaloids from Toddalia asiatica . Nat Prod Lett 1995; 6: 153-156
- 11 Sharma U, Bala M, Kumar N, Singh B, Munshi RK, Bhalerao S. Immunomodulatory active compounds from Tinospora cordifolia . J Ethnopharmacol 2012; 141: 918-926
- 12 Ahmad W, Jantan I, Kumolosasi E, Bukhari SNA. Standardized extract of Tinospora crispa stimulates innate and adaptive immune responses in Balb/c mice. Food Funct 2016; 7: 1380-1389
- 13 Ahmad W, Jantan I, Kumolosasi E, Bukhari SNA. Immunostimulatory effects of the standardized extract of Tinospora crispa on innate immune responses in Wistar Kyoto rats. Drug Des Dev Ther 2015; 9: 2961
- 14 Haque MA, Jantan I, Bukhari SNA. Tinospora species: an overview of their modulating effects on the immune system. J Ethnopharmacol 2017; 207: 67-85
- 15 Ahmad W, Jantan I, Bukhari SN. Tinospora crispa (L.) Hook. f. & Thomson: a review of its ethnobotanical, phytochemical, and pharmacological aspects. Front Pharmacol 2016; 7: 59
- 16 Huskisson EC, Scott J, Balme HW, Dieppe PA, Trapnell J, Willoughby DA. Immunostimulant therapy with levamisole for rheumatoid arthritis. Lancet 1976; 307: 393-395
- 17 Ahmad W, Jantan I, Kumolosasi E, Haque MA, Bukhari SNA. Immunomodulatory effects of Tinospora crispa extract and its major compounds on the immune functions of RAW 264.7 macrophages. Int Immunopharmacol 2018; 60: 141-151
- 18 Pierce JW, Schoenleber R, Jesmok G, Best J, Moore SA, Collins T, Gerritsen ME. Novel inhibitors of cytokine-induced IκBα phosphorylation and endothelial cell adhesion molecule expression show anti-inflammatory effects in vivo . J Biol Chem 1997; 272: 21096-21103
- 19 Wasser S. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. Appl Microbiol Biotechnol 2002; 60: 258-274
- 20 del Carmen Juárez-Vázquez M, Josabad Alonso-Castro A, García-Carrancá A. Kaempferitrin induces immunostimulatory effects in vitro . J Ethnopharmacol 2013; 148: 337-340
- 21 Mohamed SIA, Jantan I, Haque MA. Naturally occurring immunomodulators with antitumor activity: an insight on their mechanisms of action. Int Immunopharmacol 2017; 50: 291-304
- 22 Wang G, Zhu L, Yu B, Chen K, Liu B, Liu J, Qin G, Liu C, Liu H, Chen K. Exopolysaccharide from Trichoderma pseudokoningii induces macrophage activation. Carbohydr Polym 2016; 149: 112-120
- 23 Lee SJ, Rim HK, Jung JY, An HJ, Shin JS, Cho CW, Rhee YK, Hong HD, Lee KT. Immunostimulatory activity of polysaccharides from Cheonggukjang . Food Chem Toxicol 2013; 59: 476-484
- 24 Kim HS, Kim YJ, Lee HK, Ryu HS, Kim JS, Yoon MJ, Kang JS, Hong JT, Kim Y, Han SB. Activation of macrophages by polysaccharide isolated from Paecilomyces cicadae through toll-like receptor 4. Food Chem Toxicol 2012; 50: 3190-3197
- 25 Zhang CX, Dai ZR. Immunomodulatory activities on macrophage of a polysaccharide from Sipunculus nudus L. Food Chem Toxicol 2011; 49: 2961-2967
- 26 Rosenberger CM, Finlay BB. Phagocyte sabotage: disruption of macrophage signalling by bacterial pathogens. Nat Rev Mol Cell Biol 2003; 4: 385-396
- 27 Twigg HL. Macrophages in innate and acquired immunity. Semin Respir Crit Care Med 2004; 25: 21-31
- 28 Mishra K, Padwad Y, Jain M, Karan D, Ganju L, Sawhney R. Aqueous extract of Rhodiola imbricata rhizome stimulates proinflammatory mediators via phosphorylated IκB and transcription factor nuclear factor-κB. Immunopharmacol Immunotoxicol 2006; 28: 201-212
- 29 Hoshino K, Takeuchi O, Kawai T, Sanjo H, Ogawa T, Takeda Y, Takeda K, Akira S. Cutting edge: toll-like receptor 4 (TLR4)-deficient mice are hyporesponsive to lipopolysaccharide: evidence for TLR4 as the Lps gene product. J Immunol 1999; 162: 3749-3752
- 30 Li C, Meng M, Chen D, Wang Z, Han H, Chen H, Zhang J, Li X. The immunostimulatory effect of novel immunostimulator CH2b with a thiazolidin-4-one ring on the functions of LPS-activated RAW 264.7 macrophages in vitro . Int Immunopharmacol 2013; 17: 698-703
- 31 Tham CL, Liew CY, Lam KW, Mohamad AS, Kim MK, Cheah YK, Zakaria ZA, Sulaiman MR, Lajis NH, Israf DA. A synthetic curcuminoid derivative inhibits nitric oxide and proinflammatory cytokine synthesis. Eur J Pharmacol 2010; 628: 247-254
- 32 Bertholet S, Tzeng E, Felley-Bosco E, Mauel J. Expression of the inducible NO synthase in human monocytic U937 cells allows high output nitric oxide production. J Leukoc Biol 1999; 65: 50-58
- 33 Haque MA, Jantan I, Harikrishnan H. Zerumbone suppresses the activation of inflammatory mediators in LPS-stimulated U937 macrophages through MyD88-dependent NF-κB/MAPK/PI3K-Akt signaling pathways. Int Immunopharmacol 2018; 55: 312-322
- 34 Kopp EB, Ghosh S. NF-κB and Rel proteins in innate immunity. Adv Immunol 1995; 58: 1-27
- 35 Phan HH, Cho K, Sainz-Lyon KS, Shin S, Greenhalgh DG. CD14-dependent modulation of NF-kappaB alternative splicing in the lung after burn injury. Gene 2006; 371: 121-129
- 36 Kefaloyianni E, Gaitanaki C, Beis I. ERK1/2 and p 38-MAPK signalling pathways, through MSK1, are involved in NF-κB transactivation during oxidative stress in skeletal myoblasts. Cell Signal 2006; 18: 2238-2251
- 37 Smits HA, de Vos NM, Wat JW, van der Bruggen T, Verhoef J, Nottet HS. Intracellular pathways involved in TNF-α and superoxide anion release by Aβ (1–42)-stimulated primary human macrophages. J Neuroimmunol 2001; 115: 144-151
- 38 Manning BD, Cantley LC. AKT/PKB signaling: navigating downstream. Cell 2007; 129: 1261-1274
- 39 Bai D, Ueno L, Vogt PK. Akt-mediated regulation of NFκB and the essentialness of NFκB for the oncogenicity of PI3K and Akt. Int J Cancer 2009; 125: 2863-2870
- 40 Laird MH, Rhee SH, Perkins DJ, Medvedev AE, Piao W, Fenton MJ, Vogel SN. TLR4/MyD88/PI3K interactions regulate TLR4 signaling. J Leukoc Biol 2009; 85: 966-977
- 41 Xie Q, Kashiwabara Y, Nathan C. Role of transcription factor NF-kappa B/Rel in induction of nitric oxide synthase. J Biol Chem 1994; 269: 4705-4708
- 42 Bonaiuto C, McDonald PP, Rossi F, Cassatella MA. Activation of nuclear factor-κB by β-amyloid peptides and interferon-γ in murine microglia. J Neuroimmunol 1997; 77: 51-56
- 43 Gorina R, Font-Nieves M, Marquez-Kisinousky L, Santalucia T, Planas AM. Astrocyte TLR4 activation induces a proinflammatory environment through the interplay between MyD88-dependent NFkappaB signaling, MAPK, and Jak1/Stat1 pathways. Glia 2011; 59: 242-255
- 44 Hamid HA, Yusoff MM, Liu M, Karim MR. α-Glucosidase and α-amylase inhibitory constituents of Tinospora crispa: Isolation and chemical profile confirmation by ultra-high performance liquid chromatography-quadrupole time-of-flight/mass spectrometry. J Funct Foods 2015; 16: 74-80
- 45 Harikrishnan H, Jantan I, Haque MA, Kumolosasi E. Anti-Inflammatory effects of hypophyllanthin and niranthin through downregulation of NF-κB/MAPKs/PI3K-Akt signaling pathways. Inflammation 2018; 41: 984-995
- 46 Okoko T, Oruambo IF. Inhibitory activity of quercetin and its metabolite on lipopolysaccharide-induced activation of macrophage U937 cells. Food Chem Toxicol 2009; 47: 809-812
- 47 Aluwi MFFM, Rullah K, Haque MA, Yamin BM, Ahmad W, Amjad MW, Leong SW, Fahmizar NA, Jalil J, Abas F. Suppression of PGE2 production via disruption of MAPK phosphorylation by unsymmetrical dicarbonyl curcumin derivatives. Med Chem Res 2017; 26: 3323-3335