Functional analysis of the antioxidant activity of immune-challenged Bombyx mori hemolymph extracts in the human epithelial Caco-2 cell line

  • Kim, Seong Ryul (Sericultural and Apicultural Materials Division, National Institute of Agricultural Sciences, RDA) ;
  • Kim, Kee-Young (Sericultural and Apicultural Materials Division, National Institute of Agricultural Sciences, RDA) ;
  • Kim, Seong-Wan (Sericultural and Apicultural Materials Division, National Institute of Agricultural Sciences, RDA) ;
  • Park, Seung-Won (Department of Biomedical Science, Daegu Catholic University)
  • Received : 2020.02.24
  • Accepted : 2020.03.13
  • Published : 2020.03.31


Humans use insects as food and traditional medicine for many years. Hemolymph is the circulating fluid of insects and is a key component of their immune system. However, limited information is available regarding hemolymph identification, development, and differentiation, as well as the related cellular immune responses. In a previous study, hemolymph extracts prepared from Bombyx mori larvae were found to exert anti-inflammatory effects. In this study, we aimed to identify and compare the antioxidant activity of immune-challenged and unchallenged B. mori hemolymph extracts in vitro. For this purpose, human epithelial Caco-2 cells were first exposed to oxidative stress and then treated with various concentrations and incubation times of either immune-challenged or unchallenged B. mori hemolymph extracts. Next, we determined the effect of treatment on the relative expression of GPX-1, SOD-1, and SOD-2 antioxidant marker genes. We found that the expression rates of the three marker genes were markedly higher at a immune-challenged hemolymph extract concentration of 80 ppm compared to those at other concentrations, and the antioxidant effects were enhanced after treatment for 48 hr. Thus, B. mori hemolymph extracts showed antioxidant activity within the limited time and dose. Especially, the immune-challenged B. mori hemolymph extracts showed higher the antioxidant activities than unchallenged one. The activity of silkworm hemolymph extracts could facilitate the development of new types of functional foods, feed additives, and biomaterials with antioxidant properties.


Supported by : Rural Development Administration


  1. Aono A, Hazama M, Notoya K, Taketomi S, Yamasaki H, Tsukuda R, et al. (1995) Potent ectopic bone-inducing activity of bone morphogenetic protein-4/7 heterodimer. Biochem Bioph Res Co 210, 670-677.
  2. Balakrishnan D, Kandasamy D, Nithyanand P (2014) A review on antioxidant activity of marine organisms. Int J Chem Tech Res 6(7), 3431-3436.
  3. Barbehenn RV (2002) Gut-based antioxidant enzymes in a polyphagous and a graminivorous grasshopper. J Chem Ecol 28, 1329-1347.
  4. Baynes JW (1991) Role of oxidative stress in development of complications in diabetes. Diabetes 40(4), 405-412.
  5. Chen Y, Riese MJ, Killinger MA, Hoffmann FM (1998) A genetic screen for modifiers of Drosophila decapentaplegic signaling identifies mutations in punt, Mothers against dpp and the BMP-7 homologue, 60A. Development 125, 1759-1768.
  6. Chew BP, Park JS (2004) Carotenoid action on the immune response. J Nutr 134(1), 257S-261S.
  7. Dhinaut J, Balourdet A, Teixeira M, Chogne M, Moret Y (2017) A dietary carotenoid reduces immunopathology and enhances longevity through an immune depressive effect in an insect model. Sci Rep 7, 12429.
  8. Felton GW, Summers CB (1995) Antioxidant systems in insects. Arch Insect Biochem Physiol 29(2), 187-197.
  9. Fridovich I (1978) The biology of oxygen radicals. Science 201(4359), 875-880.
  10. Kim SR, Park JW, Kim S, Kim SB, Jo Y, Kim KY, et al. (2018) Antibacterial effects of two ceropin type peptide isolated from the silkworm against Salmonella species. Int J Indust Entomol 37(2), 95-99.
  11. Kim SR, Hong SJ, Choi K, Kim SW, Jeong ST, Park SW (2019) Antibacterial and anti-inflammatory activities of the immune-challenged silkworm (Bombyx mori) hemolymph with Lactobacillus cell wall extracts. Entomol Res 49, 354-362.
  12. Kim YI, Choi K, Kim SR, Goo TW, Park SW (2017) Bombyx mori hemocyte extract has anti-inflammatory effects on human phorbol myristate acetate-differentiated THP-1 cells via TLR4-mediated suppression of the NF-${\kappa}B$ signaling pathway. Mol Med Rep 16, 4001-4007.
  13. Kunnapuu J, Shimmi O (2010) Evolutional imprints on the sequences of BMP2/4/DPP type proteins. Fly (Austin) 4, 21-23.
  14. Landis GN, Tower J (2005) Superoxide dismutase evolution and life span regulation. Mech Ageing Dev 126(3), 365-379.
  15. Lettieri-Barbato D, Tomei F, Sancini A, Morabito G, Serafini M (2013) Effect of plant foods and beverages on plasma non-enzymatic antioxidant capacity in human subjects: a meta-analysis. Br J Nutr 109, 1544-1556.
  16. Magrone T, Perez de Heredia F, Jirillo E, Morabito G, Marcos A, Serafini M (2013) Functional foods and nutraceuticals as therapeutic tools for the treatment of diet-related diseases. Can J Physiol Pharmacol 91, 387-396.
  17. Mason JB, Black R, Booth SL, Brentano A, Broadbent B, Connolly, et al. (2018) Fostering strategies to expand the consumption of edible insects: the value of a tripartite coalition between academia, industry, and government. Curr Dev Nutr 2, 1-5.
  18. Mattia CD, Battista N, Sacchetti G, Serafini M (2019) Antioxidant activities in vitro of water and liposoluble extracts obtained by different species of edible insects and invertebrates. Front Nutr 6, 106.
  19. Mittapalli O, Neal JJ, Shukle RH (2007) Antioxidant defense response in a galling insect. Proc Natl Acad Sci U S A 104(6), 1889-1894.
  20. Mookherjee N, Brown KL, Bowdish DM, Doria S, Falsafi R, Hokamp K, et al. (2006) Modulation of the TLR-mediated inflammatory response by the endogenous human host defense peptide LL-37. J Immunol 176, 2455-2464.
  21. Morath S, Geyer A, Hartung T (2001) Structure-function relationship of cytokine induction by lipoteichoic acid from Staphylococcus aureus. J Exp Med 193, 393-397.
  22. Munford RS, Pugin J (2001) Normal responses to injury prevent systemic inflammation and can be immunosuppressive. Am J Respir Criti Care Med 163, 316-321.
  23. Oghenesuvwe EE, Paul C (2019) Edible insects bio-actives as antioxidants: Current status and perspectives. J Complement Med Res 10(2), 89-102.
  24. Pardini RS (1995) Toxicity of oxygen from naturally occurring redoxactive pro-oxidants. Arch Insect Biochem Physiol 29(2), 101-118.
  25. Pimentel D, Pimentel M (2003) Sustainability of meat-based and plant-based diets and the environment. Am J Clin Nutr 78(Suppl.).
  26. Serafini M, Bugianesi R, Maiani M, Valtuena S, De Santis S, Crozier A (2003) Plasma antioxidants from chocolate. Nature 424, 1013.
  27. Serafini M, Miglio C, Peluso I, Petrosino T (2011) Modulation of plasma non-enzimatic antioxidant capacity (NEAC) by plant foods: the role of polyphenols. Curr Top Med Chem 11, 1821-1846
  28. Sosa V, Moline T, Somoza R, Paciucci R, Kondoh H, LLeonart ME (2013) Oxidative stress and cancer: an overview. Ageing Res Rev 12(1), 376-390.
  29. Van Huis A, Van Itterbeek J, Klunder H, Mertens E, Halloran A, Muir G, et al. (2013) Edible Insects. Future Prospects for Food and Feed Security. Vol. 171. Pp. 60-64. Food and Agriculture Organization of the United Nations, Rome.
  30. Yoon KY, Kim KJ, Youn HS, Oh SR, Lee BY (2015) Brazilin suppresses inflammation via the down-regulation of IRAK4 in LPS-stimulated Raw264.7 macrophage. J Food Nutr Res 3(9), 575-580.
  31. Zielinska E, Karas M, Jakubczyk A (2017) Antioxidant activity of predigested protein obtained from a range of farmed edible insects. Int J Food Sci Tech 52(2), 306-312.
  32. Zhang Y, Yang F, Jamali MA, Peng Z (2016) Antioxidant enzyme activities and lipid oxidation in rape (Brassica campestris L.) bee pollen added to salami during processing. Molecules 21(11).