Inhibition of Human Neutrophil Functional Responsiveness by Cryptolepis buchanani Extract

Main Article Content

Payong Wanikiat
Ampai Panthong
VIchai Reutrakul

Abstract

            Cryptolepis buchanani Roem. & Schult. (Asclepiadaceae), a climbing tree and commonly known as “Thao En On” in Thailand, is widely used in traditional folk medicine in Southeast Asia. In Thailand, the stem of this plant is traditionally used for the treatment of inflammation, including arthritis and muscle and joint pain. Alcoholic extract from stem of C. buchanani was found to possess anti-inflammatory activities in animal models. This study aimed to investigate the in vitro effects of C. buchanani extract on human neutrophil functional responsiveness in order to elucidate the underlying cellular mechanisms of its acute anti-inflammatory effects. Human neutrophil functional responsiveness was determined by measuring N-formyl-methionyl-leucyl-phenylalanine (fMLP)-induced chemotaxis, superoxide anion generation (SAG), and release of myeloperoxidase (MPO) and elastase. Apoptosis was assessed morphologically and by flow cytometry. Neutrophil viability was assessed by trypan blue exclusion and 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)- 2H-tetrazolium-5-carboxanilide (XTT) cytotoxic assay. Although C. buchanani extract did not affect neutrophil viability and apoptosis, treatment of neutrophils with the herbal extract concentration-dependently inhibited fMLP-induced chemotaxis, SAG, and MPO release. In addition, neutrophil apoptosis was slightly stimulated by the herbal extract. In conclusion, these findings suggest that the anti-inflammatory property of C. buchanani extract is mediated, in part, by inhibition of neutrophil functional responsiveness. The results lend support to the effectiveness of C. buchanani in combating inflammation. However, further investigations are required to fully identify the biologically active compound(s) of C. buchanani and to define the underlying molecular and cellular mechanisms of action involved.

Article Details

Section
Research Articles

References

1. Sharma D, Sahu AN, Mujeeb M, Bharti A, Sharma A, Tripathi RK et al. Development of pharmacognostical profile of Cryptolepis buchanani Roem. & Schult. Int J Pharm Pharm Sci. 2012;4(2):615-8.
2. Tayung K, Saikia N. Cryptolepis buchanani - A less-known medicinal plant used in bone fracture. Indian J Tradit Know. 2003;2(4):371-4.
3. Panthong A, Kanjanapothi D, Talor WC. Ethnobotanical review of medicinal plants from Thai traditionnal books, Part I: Plants with anti-inflammatory, anti-asthmatic and antihypertensive properties. J Ethnopharmacol. 1986 Dec;18(3): 213-28.
4. Laupattarakasem P, Houghton PJ, Hoult JR, Itharat A. An evaluation of the activity related to inflammation of four plants used in Thailand to treat arthritis. J Ethnopharmacol. 2003 Apr;85(2-3):207-15.
5. Laupattarakasem P, Wangsrimongkol T, Surarit R, Hahnvajanawong. In vitro and in vivo anti-inflammatory potential of Cryptolepis buchanani. J Ethnopharmacol. 2006 Dec 6;108(3):349-54.
6. Hanprasertpong N, Teekachunhatean S, Chaiwongsa R, Ongchai S, Kunanusorn P, Sangdee C, et al. Analgesic, anti-inflammatory, and chondroprotective activities of Cryptolepis buchanani extract: in vitro and in vivo studies. Biomed Res Int. 2014;2014:978582.
7. Sittiwet C, Puangpronpitag D. Anti-bacterial activity of Cryptolepis buchanani aqueous extract. Int J Biol Chem. 2009;3(2):90-4.
8. Viniyaka KS, Prashith Ketkuda TR, Mallikarjun N, Sateesh VN. Anti-dermatophyte of Cryptolepis buchanani Roem. & Schult. Pharmacogn J. 2010 Mar;2(7):170-2.

9. Hanprasertpong N, Panthong A, Sangdee C, Kunanusorn P, Kasitanon N, Lhieochaiphant S, et al. Cryptolepis buchanani oil formulation versus indomethacin solution in topical therapy for osteoarthritis of the knee: A randomized controlled trial. J Herb Med. 2017 Mar;7:18-26.
10. Kolaczkowska E, Kubes P. Neutrophil recruitment and function in health and inflammation. Nat Rev Immunol. 2013 Mar;13(3):159-75.
11. Cuzzocrea S, Riley DP, Caputi AP, Salvemini D. Antioxidant therapy: a new pharmacological approach in shock, inflammation, and ischaemia/reperfusion injury. Pharmacol Rev. 2001 Mar;53(1):135-59.
12. Walker A, Ward C, Taylor EL, Dransfield I, Hart SP, Haslett C, et al. Regulation of neutrophil apoptosis and removal of apoptotic cells. Curr Drug Targets Inflamm Allergy. 2005 Aug;4(4):447-54.
13. Haslett C. Granulocyte apoptosis and its role in the resolution and control of lung inflammation. Am J Respir Crit Care Med. 1999 Nov;160(5 Pt 2):S5-11.
14. Gilroy DW, Lawrence T, Perretti M, Rossi AG. Inflammatory resolution: new opportunities for drug discovery. Nat Rev Drug Discov. 2004 May;3(5)::401-16.
15. Kankaanranta H, Moilanen E, Vapatalo H. Effects of non-steroidal anti-inflammatory drugs on polymorphonuclear leukocyte functions in vitro: focus on fenamates. Naunyn Schmiedebergs Arch Pharmacol. 1994 Dec;350(6):685-91.
16. Wipke BT, Allen PM. Essential role of neutrophils in the initiation and progression of a murine model of rheumatoid arthritis. J Immunol. 2001 Aug 1; 167(3):1601-8.
17. Bombini G, Canetti C, Rocha FA, Cunba FQ. Tumour necrosis factor-alpha mediates neutrophil migration to the knee synovial cavity during immune inflammation. Eur J Pharmacol. 2004 Aug 2;496(1-3:197-204.
18. Wright HL, Moots RJ, Edwards SW. The multifactorial role of neutrophils in rheumatoid arthritis. Nat Rev Rheumatol. 2014 Oct;10(10):593-601.
19. Chen W, Wang Q, Ke Y, Lin J. Neutrophil function in an inflammatory milieu of rheumatoid arthritis. J Immunol Res. 2018 Dec 3;2018:8549329.
20. Kandahari A, Yang X, Dighe A, Pan D, Cui Q. Recognition of immune response for the early diagnosis and treatment of osteoarthritis. J Immunol Res. 2015; 2015;2015:192415.
21. Kraan MC, Koster BM, Elferink JG, Post WJ, Breedveld FC, Tak PP. Inhibition of neutrophil migration soon after initiation of treatment with leflunomide or methotrexate in patients with rheumatoid arthritis: Finding in a prospective, randomized, double-blind clinical trial in fifteen patients. Arthritis Rheum. 2000 Jul;43(7):1488-95.
22. den Broeder AA, Wanten GJ, Oyen WJ, Naber T, van Riel PL, Barrera P. Neutrophil migration and production of reactive oxygen species during treatment with a fully human anti-tumor necrosis factor-α monoclonal antibody in patients with rheumatoid arthritis. J Rheumatol. 2003 Feb;30(2):232-7.
23. Cavin A, Hostettmann K, Dyatmyko W, Pototterat O. Antioxidant and lipophilic constituents of Tinospora crispa. Planta Med. 1998 Jun;64(5):393-6.
24. Roehm NW, Rodgers GH, Hatfield SM, Glasebrook AL. An improved colori-metric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J Immunol Methods. 1991 Sep 13;142(2):257-65.
25. Wanikiat P, Woodward DF, Armstrong RA. Investigation of the role of nitric oxide and cyclic GMP in both the activation and inhibition of human neutrophils. Br J Pharmacol. 1997 Nov;122(6):1135-45.
26. Talpain E, Armstrong RA, Coleman RA, Vardey CJ. Characterisation of the PGE receptor subtype mediating inhibition of superoxide production in human neutrophils. Br J Pharmacol. 1995 Apr;114(7):1459-65.
27. Suzuki K, Ota H, Sasagawa S, Sakatani T, fujikura T. Assay method for myelo-peroxidase in human polymorphonuclear leukocytes. Anal Biochem. 1983 Jul 15;132(2):345-52.
28. Barrett AJ. Leukocyte elastase. Method Enzymol. 1981;80 Pt C:581-8.
29. Ward C, Chilvers ER, Lawson MF, Pryde JG, Fujihara S, Farrow SN, et al. NF-kappaB activation is a critical regulator of human granulocyte apoptosis in vitro. J Biol Chem. 1999 Feb 12;274(7):4309-18.
30. Ward C, Dransfield I, Chilvers ER, Haslett C, Rossi AG. Pharmacological manipulation of granulocyte apoptosis: potential therapeutic targets. Trends Pharmacol Sci. 1999 Dec;20(12):503-9.
31. Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, Van Oers MH. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood. 1994 Sep 1;84(5):1415-20.
32. Sabroe I, Prince LR, Jones EC, Horsburgh MJ, Foster SJ, Vogel SN, et al. Selective roles for Toll-like receptor (TLR)2 and TLR4 in the regulation of neutrophil activation and life span. J Immunol. 2003 May 15;170(10)::5268-75.
33. Karlsson A, Nixon JB, McPhail LC. Phorbol myristate acetate induces neutrophil NADPH-oxidase activity by two separate signal transduction pathways: dependent or independent of phosphatidyl-inositol 3-kinase. J Leukoc Biol. 2000 Mar;67(3): 396-404.
34. Ashwini SK, Kiran R, Soumyn KV, Sudharshan SJ, Prashith Kekuda TR, Vinayaka KS, et al. Insecticidal and in vitro antioxidant potency of extracts of Cryptolepis buchanani Roem. & Schult. Int J Ph Sci. 2010;2(1);418-25.
35. Ramos CL, Pou S, Rosen GM. Effects of anti-inflammatory drugs on myelo-peroxidase-dependent hydroxyl radical generation by human neutrophils. Biochem Pharmacol. 1995 Apr 18;49(8):1079-84.
36. Meagher LC, Cousin JM, Seckl JR, Haslett C. Opposing effects of gluco-corticoids on the rate of apoptosis in neutrophilic and eosinophlic granulocytes. J Immunol. 1996 Jun 1;156(11):4422-8.