Anti-Inflammatory Properties of Piperlactam S: Modulation of Complement 5a-Induced Chemotaxis and Inflammatory Cytokines Production in Macrophages

Abstract

Macrophages infiltrate tissues in response to chemoattractants including complement 5a (C5a). Infiltrating macrophages clear microorganisms but also can cause tissue damage. We hypothesized that prevention of macrophages from excessive recruitment into infected sites may underlie the anti-inflammatory effects of piperlactam S, an alkaloid isolated from Piper kadsura (Choisy) Ohwi. To test this hypothesis, chemotactic migration of RAW264.7 macrophages was induced by C5a and the effects of piperlactam S were studied. The results showed that piperlactam S (1 - 30 μM) concentration-dependently suppressed C5a-induced migration across a fibrinogen-coated barrier with an IC₅₀ of 4.5 ± 0.3 μM. At 30 μM, piperlactam S inhibited chemotaxis by more than 95 % and also decreased phagocytosis by 25 % without reducing macrophage viability and adherent capacity. Furthermore, piperlactam S treated cells adhered but failed to spread and elongate as in control cells. Finally, piperlactam S inhibited the C5a-stimulated release of tumor necrosis factor-α and interleukin-1β. We conclude that retardation of macrophage recruitment by interfering with the migration process and suppression of cytokines production might underlie the potential usefulness of piperlactam S as an anti-inflammatory agent.

Key words

Piper kadsura (Choisy) Ohwi - Piperaceae - piperlactam S - anti-inflammation - chemotaxis - cell spreading - cytokines

References

• 1 Chung C Y, Funamoto S, Firtel R A. Signaling pathways controlling cell polarity and chemotaxis.  Trends Biochem Sci. 2001;  26 557-66
• 2 Paczkowski N J, Finch A M, Whitmore J, Short A J, Wong A K, Monk P N, Cain S A, Fairlie D P, Taylor S M. Pharmacological characterization of antagonists of the C5a receptor.  Br J Pharmacol. 1999;  128 1461-6
• 3 Czermak B J, Sarma V, Pierson C L, Warner R L, Huber-Lang M, Bless N M, Schmal H, Friedl H P, Ward P A. Protective effects of C5a blockade in sepsis.  Nat Med. 1999;  5 788-92
• 4 Heller T, Hennecker M, Baumann U, Gessner J E, Zu Vilsendorf A S, Baensch M, Boulay F, Kola F, Klos A, Bautsch W, Kohl J. Selection of a C5a receptor antagonist from phage libraries attenuating the inflammatory response in immune complex disease and ischemia/reperfusion injury.  J Immunol. 1999;  163 985-94
• 5 Han G Q, Dai P, Xu L, Ma J, Ki C L, Zheng Q T. PAF inhibitors: Neolignans from Piper kadsura .  Planta Med. 1990;  56 583-4
• 6 Kuo Y C, Yang N S, Chou C J, Lin L C, Tsai W J. Regulation of cell proliferation, gene expression, production of cytokines, and cell cycle progression in primary human T lymphocytes by piperlactam S isolated from Piper kadsura .  Mol Pharmacol. 2000;  58 1057-66
• 7 Jones G E. Cellular signaling in macrophage migration and chemotaxis.  J Leuko Biol. 2000;  68 593-602
• 8 Fine R, Shaw J O, Rogers W R. Effects of C5a on baboon alveolar macrophage migration.  Am Rev Respir Dis. 1981;  123 110-4
• 9 Shen Y C, Chou C J, Chiou W F, Chen C F. Anti-inflammatory effects of the partially purified extract of Radix Stephaniae tetrandrae: comparative studies of its active principles tetrandrine and fanchinoline on human polymorphonuclear leukocyte functions.  Mol Pharmacol. 2001;  60 1083-90
• 10 Shen Y C, Chen C F, Wang S Y, Sung Y J. Impediment to calcium influx and reactive oxygen production accounts for the inhibition of neutrophil Mac-1 up-regulation and adhesion by tetrandrine.  Mol Pharmacol. 1999;  55 186-93
• 11 Del Rio M, Ruedas G, Medina S, Victor V M, De la Fuente M. Improvement by several antioxidants of macrophage function in vitro .  Life Sci. 1998;  63 871-81
• 12 Warskulate U, Zhang F, Haussinger D. Modulation of phagocytosis by anisoosmolarity and betaine in rat liver macrophages (Kupffer cells) and RAW264.7 mouse macrophages.  FEBS Letters. 1996;  391 287-92
• 13 Sanchez-Madrid F, Del Pozo M A. Leukocyte polarization in cell migration and immune interactions.  EMBO J. 1999;  18 501-11
• 14 Haynes D R, Harkin D G, Bignold L P, Hutchens M J, Taylor S M, Fairlie D P. Inhibition of C5a-induced neutrophil chemotaxis and macrophage cytokine production in vitro by a new C5a receptor antagonist.  Biochem Pharmacol. 2000;  60 729-33
• 15 Kravchenko V V, Pan Z, Han J, Herbert J M, Ulevitch R J, Ye R D. Platelet-activating factor induces NF-kappa B activation through a G protein-coupled pathway.  J Biol Chem. 1995;  270 14 928-34
• 16 Katz Y, Nadiv O, Beer Y. Interleukin-17 enhances tumor necrosis factor alpha-induced synthesis of interleukins-1, -6, and -8 in skin and synovial fibroblasts: a possible role as a ”fine-tuning cytokine” in inflammation processes.  Arth Rheu. 2001;  44 2176-84
• 17 Pearce G J, Chikanza I C. Targeting tumor necrosis factor in the treatment of rheumatoid arthritis.  Biodrugs. 2001;  15 139-49
• 18 Tominaga K, Alstergren P, Kurita H, Matsukawa A, Fukuda J, Kopp S. Interleukin-1beta in antigen-induced arthritis of the rabbit temporomandibular joint.  Arch Oral Biol. 2001;  46 539-44
• 19 Sasaki M, Kashima M, Ito T, Watanabe A, Izumiyama N, Sano M, Kagaya M, Shioya T, Miura M. Differential regulation of metalloproteinase production, proliferation and chemotaxis of human lung fibroblasts by PDGF, interleukin-1beta and TNF-alpha.  Med Inflamm. 2000;  9 155-60
• 20 Ojo-Amaize E A, Kapahi P, Kakkanaiah V N, Takahashi T, Shalom-Barak T, Cottam H B, Adesomoju A A, Nchekwube E J, Oyemade O A, Karin M, Okogun J I. Hypoestoxide, a novel anti-inflammatory natural diterpene, inhibits the activity of IkappaB kinase.  Cellular Immunol. 2001;  209 149-57