Abstract
Tragia benthamii (TBM) commonly called the climbing nettle is a tropical plant claimed to have numerous anti inflammatory effects in sub Saharan African ethnomedicine which lacks scientific evidence. Aqueous extracts of TBM were further prepurified on a RP-C18 parked solid phase system to obtain 20% aqueous fraction. This fraction was enzymatically and chemically analyzed (by MALDI TOF MS and MS/MS) to contain interesting low molecular weight cysteine-rich stable peptides within the range of 2.5–3.2 KDa. The 20% aqueous fraction was further tested in vivo using carrageenan-induced foot edema (acute inflammation) in seven-day old chicks with diclofenac as reference drug. The cytotoxicity of this active fraction was investigated using the brine shrimp lethality assay. The brine shrimp cytotoxicity assay produced LC50 above 1000 μg/mL. Pretreatment with the TBM extract (30–300 mg/kg, i.p) dose dependently (P<0.01) reduced foot edema with maximal inhibition of 0.253 ± 0.180 (84.3%) at 300 mg/kg body weight, which was comparable to that of diclofenac with inhibition (P<0.05) of 0.410 ± 0.271 (74.5%) at 10 mg/kg body weight. The study has therefore shown for the first time, the detection of cysteine-rich biologically active peptides in T. benthamii and the stable peptide extracts from this ethnomedicinal plant, which is not toxic to Artemia salina, exhibits anti inflammatory activity in a chick in vivo model. This may provide scientific evidence for its use in the treatment of inflammation and pain in traditional medicine. Further in-depth vivo and in vitro studies will be required to investigate its anti inflammatory activity including effect on HUVEC-TERT, the possible inhibition of ICAM-1 surface expression and the mechanism of the anti inflammatory effect.
Acknowledgement
Authors wish to thank Associate Professor Christian W. Gruber of the center for physiology and pharmacology, mass spectrometry facility, Medical University of Vienna, Austria for access to the mass spectrometry laboratory during the experiment.
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Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
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Research funding: Authors declare no funding was received for this work.
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Conflict of interest statement: The authors declare no conflicts of interest regarding this article.
References
1. Reddy, BS, Rao, NR, Vijeepallam, K, Pandy, V. Phytochemical, pharmacological and biological profiles of Tragia species (family: Euphorbiaceae). Afr J Tradit, Complementary Altern Med 2017;14:105–12. https://doi.org/10.21010/ajtcam.v14i3.11.Search in Google Scholar PubMed PubMed Central
2. Balogun, O, Oladosu, I, Liu, Z. Isolation of 2, 5-dithia-3, 6-diazabicyclo [2.2. 1] heptane and GC-MS analysis of silylated extract from Tragia benthamii. IFE J Sci 2020;22:075–80. https://doi.org/10.4314/ijs.v22i2.7.Search in Google Scholar
3. Gillespie, LJ, Cardinal-McTeague, WM, Wurdack, KJ. Monadelpha (Euphorbiaceae, Plukenetieae), a new genus of Tragiinae from the Amazon rainforest of Venezuela and Brazil. PhytoKeys 2020;169:119. https://doi.org/10.3897/phytokeys.169.59244.Search in Google Scholar PubMed PubMed Central
4. Narasimhan, S. Pharmacological potential of the stinging plant Tragia species: a review. Phcog J 2021;13:278–84. https://doi.org/10.5530/pj.2021.13.37.Search in Google Scholar
5. Oladosu, I, Balogun, S, Ademowo, G. Phytochemical screening, antimalarial and histopathological studies of Allophylus africanus and Tragia benthamii. Chin J Nat Med 2013;11:371–6. https://doi.org/10.1016/s1875-5364(13)60054-0.Search in Google Scholar
6. Fred-Jaiyesimi, AA, Ajibesin, KK. Ethnobotanical survey of toxic plants and plant parts in Ogun State, Nigeria. Int J Green Pharm 2012;6:174–9. https://doi.org/10.4103/0973-8258.104926.Search in Google Scholar
7. Grivennikov, SI, Greten, FR, Karin, M. Immunity, inflammation, and cancer. Cell 2010;140:883–99. https://doi.org/10.1016/j.cell.2010.01.025.Search in Google Scholar PubMed PubMed Central
8. Khandia, R, Munjal, A. Interplay between inflammation and cancer. Adv Protein Chem Struct Biol 2020;119:199–245. https://doi.org/10.1016/bs.apcsb.2019.09.004.Search in Google Scholar PubMed
9. Esch, T, Stefano, GB, Ptacek, R, Kream, RM. Emerging roles of blood-borne intact and respiring mitochondria as bidirectional mediators of pro-and anti-inflammatory processes. Med Sci Mon Int Med J Exp Clin Res 2020;26:e924337-1. https://doi.org/10.12659/msm.924337.Search in Google Scholar PubMed PubMed Central
10. Tam, JP, Wang, S, Wong, KH, Tan, WL. Antimicrobial peptides from plants. Pharmaceuticals 2015;8:711–57. https://doi.org/10.3390/ph8040711.Search in Google Scholar PubMed PubMed Central
11. Kumari, G, Wong, KH, Serra, A, Shin, J, Yoon, HS, Sze, SK, et al.. Molecular diversity and function of jasmintides from Jasminum sambac. BMC Plant Biol 2018;18:1–13. https://doi.org/10.1186/s12870-018-1361-y.Search in Google Scholar PubMed PubMed Central
12. Koehbach, J, Attah, AF, Berger, A, Hellinger, R, Kutchan, TM, Carpenter, EJ, et al.. Cyclotide discovery in Gentianales revisited–identification and characterization of cyclic cystine-knot peptides and their phylogenetic distribution in Rubiaceae plants. Biopolymers 2013;100:438–52. https://doi.org/10.1002/bip.22328.Search in Google Scholar PubMed PubMed Central
13. Kini, SG, Nguyen, PQ, Weissbach, S, Mallagaray, A, Shin, J, Yoon, HS, et al.. Studies on the chitin binding property of novel cysteine-rich peptides from Alternanthera sessilis. Biochemistry 2015;54:6639–49. https://doi.org/10.1021/acs.biochem.5b00872.Search in Google Scholar PubMed
14. Kini, SG, Wong, KH, Tan, WL, Xiao, T, Tam, JP. Morintides: cargo-free chitin-binding peptides from Moringa oleifera. BMC Plant Biol 2017;17:1–13. https://doi.org/10.1186/s12870-017-1014-6.Search in Google Scholar PubMed PubMed Central
15. Kumari, G, Serra, A, Shin, J, Nguyen, PQ, Sze, SK, Yoon, HS, et al.. Cysteine-rich peptide family with unusual disulfide connectivity from Jasminum sambac. J Nat Prod 2015;78:2791–9. https://doi.org/10.1021/acs.jnatprod.5b00762.Search in Google Scholar PubMed
16. Bhardwaj, G, Mulligan, VK, Bahl, CD, Gilmore, JM, Harvey, PJ, Cheneval, O, et al.. Accurate de novo design of hyperstable constrained peptides. Nature 2016;538:329–35. https://doi.org/10.1038/nature19791.Search in Google Scholar PubMed PubMed Central
17. Hellinger, R, Gruber, CW. Peptide-based protease inhibitors from plants. Drug Discov Today 2019;24:1877–89. https://doi.org/10.1016/j.drudis.2019.05.026.Search in Google Scholar PubMed PubMed Central
18. Hellinger, R, Koehbach, J, Soltis, DE, Carpenter, EJ, Wong, GK-S, Gruber, CW. Peptidomics of circular cysteine-rich plant peptides: analysis of the diversity of cyclotides from Viola tricolor by transcriptome and proteome mining. J Proteome Res 2015;14:4851–62. https://doi.org/10.1021/acs.jproteome.5b00681.Search in Google Scholar PubMed PubMed Central
19. Retzl, B, Hellinger, R, Muratspahić, E, Pinto, ME, Bolzani, VS, Gruber, CW. Discovery of a beetroot protease inhibitor to identify and classify plant-derived cystine knot peptides. J Nat Prod 2020;83:3305–14. https://doi.org/10.1021/acs.jnatprod.0c00648.Search in Google Scholar PubMed PubMed Central
20. Srivastava, S, Dashora, K, Ameta, KL, Singh, NP, El-Enshasy, HA, Pagano, MC, et al.. Cysteine-rich antimicrobial peptides from plants: the future of antimicrobial therapy. Phytother Res 2021;35:256–77. https://doi.org/10.1002/ptr.6823.Search in Google Scholar PubMed
21. Pinto, MEF, Chan, LY, Koehbach, J, Devi, S, Gründemann, C, Gruber, CW, et al.. Cyclotides from Brazilian Palicourea sessilis and their effects on human lymphocytes. J Nat Prod 2021;84:81–90. https://doi.org/10.1021/acs.jnatprod.0c01069.Search in Google Scholar PubMed PubMed Central
22. Nguyen, PQ, Wang, S, Kumar, A, Yap, LJ, Luu, TT, Lescar, J, et al.. Discovery and characterization of pseudocyclic cystine-knot α-amylase inhibitors with high resistance to heat and proteolytic degradation. FEBS J 2014;281:4351–66. https://doi.org/10.1111/febs.12939.Search in Google Scholar PubMed
23. Wong, KH, Tan, WL, Xiao, T, Tam, JP. β-ginkgotides: hyperdisulfide-constrained peptides from Ginkgo biloba. Sci Rep 2017;7:1–13. https://doi.org/10.1038/s41598-017-06598-x.Search in Google Scholar PubMed PubMed Central
24. Tammineni, R, Gulati, P, Kumar, S, Mohanty, A. An overview of acyclotides: past, present and future. Phytochemistry 2020;170:112215. https://doi.org/10.1016/j.phytochem.2019.112215.Search in Google Scholar PubMed
25. Ainooson, G, Owusu, G, Woode, E, Ansah, C, Annan, K. Trichilia monadelpha bark extracts inhibit carrageenan-induced foot-oedema in the seven-day old chick and the oedema associated with adjuvant-induced arthritis in rats. Afr J Tradit, Complementary Altern Med 2012;9:8–16.10.4314/ajtcam.v9i1.2Search in Google Scholar
26. Chuku, L, Chinaka, N, Damilola, D. Phytochemical screening and anti-inflammatory properties of Henna leaves (Lawsonia inermis). Eur J Med Plants 2020;31:23–8. https://doi.org/10.9734/ejmp/2020/v31i1830340.Search in Google Scholar
27. Mensah, JK, Ibrahim, A, Jibira, Y. Co-extract mixture from Strophanthus hispidus (roots) and Aframomum meleguta (seeds) show phytochemical synergy in its anti-inflammatory activity. J Pharm Pharm Sci 2019;3:89–100.10.29328/journal.apps.1001019Search in Google Scholar
28. Adriany, A, Jéssica, S, Ana, O, Raimunda, S, Andreanne, V, Luan, S, et al.. Anti-inflammatory and antioxidant activity improvement of lycopene from guava on nanoemulsifying system. J Dispersion Sci Technol 2020;42:1–11. https://doi.org/10.1080/01932691.2020.1728300.Search in Google Scholar
29. Banti, CN, Hadjikakou, SK. Evaluation of toxicity with brine shrimp assay. Bio-protocol 2021;11:e3895-e. https://doi.org/10.21769/bioprotoc.3895.Search in Google Scholar
30. Ogbole, OO, Ndabai, NC, Akinleye, TE, Attah, AF. Evaluation of peptide-rich root extracts of Calliandria portoriscensis (Jacq.) Benth (Mimosaceae) for in vitro antimicrobial activity and brine shrimp lethality. BMC Compl Med Ther 2020;20:30. https://doi.org/10.1186/s12906-020-2836-6.Search in Google Scholar PubMed PubMed Central
31. Gobalakrishnan, R, Kulandaivelu, M, Bhuvaneswari, R, Kandavel, D, Kannan, L. Screening of wild plant species for antibacterial activity and phytochemical analysis of Tragia involucrata L. J Pharmaceut Anal 2013;3:460–5. https://doi.org/10.1016/j.jpha.2013.07.001.Search in Google Scholar PubMed PubMed Central
32. Gressent, F, Da Silva, P, Eyraud, V, Karaki, L, Royer, C. Pea albumin 1 subunit b (PA1b), a promising bioinsecticide of plant origin. Toxins 2011;3:1502–17. https://doi.org/10.3390/toxins3121502.Search in Google Scholar PubMed PubMed Central
33. Reinwarth, M, Nasu, D, Kolmar, H, Avrutina, O. Chemical synthesis, backbone cyclization and oxidative folding of cystine-knot peptides—promising scaffolds for applications in drug design. Molecules 2012;17:12533–52. https://doi.org/10.3390/molecules171112533.Search in Google Scholar PubMed PubMed Central
34. Eyraud, V, Karaki, L, Rahioui, I, Sivignon, C, Da Silva, P, Rahbé, Y, et al.. Expression and biological activity of the cystine knot bioinsecticide PA1b (Pea Albumin 1 Subunit b). PloS One 2013;8:e81619. https://doi.org/10.1371/journal.pone.0081619.Search in Google Scholar PubMed PubMed Central
35. Antonelli, M, Kushner, I. It’s time to redefine inflammation. FASEB J 2017;31:1787–91. https://doi.org/10.1096/fj.201601326r.Search in Google Scholar
36. Fereydouni, Z, Fard, EA, Mansouri, K, Motlagh, H-RM, Mostafaie, A. Saponins from Tribulus terrestris L. extract down-regulate the expression of ICAM-1, VCAM-1 and E-selectin in human endothelial cell lines. Int J Mol Cell Med 2020;9:73. https://doi.org/10.22088/IJMCM.BUMS.9.1.73.Search in Google Scholar PubMed PubMed Central
37. Deepa, M, Devi, PR, Karur, GD. In vivo evaluation of acute and chronic anti-inflammatory activity of ethanol leaf extract of Vitex negundo Linn. JRJOB 2014;9:64–9.Search in Google Scholar
38. Machado, GC, Abdel-Shaheed, C, Underwood, M, Day, RO. Non-steroidal anti-inflammatory drugs (NSAIDs) for musculoskeletal pain. BMJ 2021;372:n104. https://doi.org/10.1136/bmj.n104.Search in Google Scholar PubMed
39. García-Rayado, G, Navarro, M, Lanas, A. NSAID induced gastrointestinal damage and designing GI-sparing NSAIDs. Expet Rev Clin Pharmacol 2018;11:1031–43. https://doi.org/10.1080/17512433.2018.1516143.Search in Google Scholar PubMed
40. Guha, S, Majumder, K. Structural-features of food-derived bioactive peptides with anti-inflammatory activity: a brief review. J Food Biochem 2019;43:e12531. https://doi.org/10.1111/jfbc.12531.Search in Google Scholar PubMed
41. Wang, N. A promising plant defense peptide against citrus huanglongbing disease. Proc Natl Acad Sci 2021;118:e2026483118. https://doi.org/10.1073/pnas.2026483118.Search in Google Scholar PubMed PubMed Central
42. Bonam, SR, Manoharan, SK, Pandy, V, Raya, AR, Nadendla, RR, Jagadeesan, M, et al.. Phytochemical, in vitro antioxidant and in vivo safety evaluation of leaf extracts of Tragia plukenetii. Phcog J 2019;11:338–45. https://doi.org/10.5530/pj.2019.11.50.Search in Google Scholar
43. Pallie, MS, Perera, PK, Kumarasinghe, N, Arawwawala, M, Goonasekara, CL. Ethnopharmacological use and biological activities of Tragia involucrata L. Evid Base Compl Alternative Med 2020;2020:8848676. https://doi.org/10.1155/2020/8848676.Search in Google Scholar PubMed PubMed Central
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