Oral Intake of Semi-refined Carrageenan by Rats Affects Apoptosis of Lymphocytes

Document Type : Research/Original Article


1 Research Institute of Experimental and Clinical Medicine, Kharkiv National Medical University; Trinklera st. 6, 61022, Kharkiv, Ukraine

2 Department of Biochemistry, Kharkiv National Medical University; Nauky av. 4, 61022, Kharkiv, Ukraine


Introduction. The safety of generally recognized as safe food additives E407 and E407a is under rigorous debate. The aim of our research was to evaluate the effects of the orally administered food additive E407a (semi-refined carrageenan) on the viability of lymphocytes and their cell death modes.
Methods. Totally, 16 adult WAG rats were divided into two equal groups (experimental – oral intake of 140 mg/kg of E407a during 2 weeks; control – oral consumption of drinking water instead). Blood samples were used to obtain leukocyte suspensions stained with Annexin V-FITC and 7-aminoactinomycin D (7-AAD). The region of lymphocytes was analyzed after collecting data using a BD FACSCanto™ II flow cytometer.
Results. Oral administration of E407a led to a decrease in the amount of viable (Annexin V-, 7-AAD-) circulating lymphocytes. Furthermore, exposure to semi-refined carrageenan resulted in an elevated number of early apoptotic (Annexin V+, 7-AAD-) lymphocytes. Their percentage was approximately 4 times higher in rats exposed to E407a compared with the control group.
Conclusion. Our findings indicate that dietary intake of E407a promotes apoptosis of circulating lymphocytes.
Keywords: processed Eucheuma seaweed, annexin V, 7-aminoactinomycin D, flow cytometry.


  1. Golshani G, Zhang Y. Advances in immunotherapy for colorectal cancer: a review. Therapeutic Advances in Gastroenterology. January 2020. doi:10.1177/1756284820917527
  2. Alexander PG, McMillan DC, Park JH. The local inflammatory response in colorectal cancer - Type, location or density? A systematic review and meta-analysis. Cancer Treat Rev. 2020;83:101949. 
  3. Kuipers EJ, Grady WM, Lieberman D, Seufferlein T, Sung JJ, Boelens PG, et al. Colorectal cancer. Nat Rev Dis Primers. 2015;1:15065. doi: 10.1038/nrdp.2015.65.
  4. Tabung FK, Brown LS, Fung TT. Dietary patterns and colorectal cancer risk: a review of 17 years of evidence (2000-2016). Curr Colorectal Cancer Rep. 2017;13(6):440-454. doi:10.1007/s11888-017-0390-5
  5. Gultekin F, Yasar S, Gurbuz N, Ceyhan BM. Food additives of public concern for their carcinogenicty. J. Nutr. Health Food Sci. 2015;3:1-6
  6. Yermak IM, Mischchenko NP, Davydova VN, Glazunov VP, Tarbeeva DV, Kravchenko AO, et al. Carrageenans-sulfated polysaccharides from red seaweeds as matrices for the inclusion of Echinochrome. Mar Drugs. 2017;15(11):337. doi: 10.3390/md15110337.
  7. Necas J, Bartosikova L. Carrageenan: a review. Veterinarni Medicina 2013;58:187-205.
  8. Tobacman JK. Review of harmful gastrointestinal effects of carrageenan in animal experiments. Environ Health Perspect. 2001;109(10):983–994. doi:10.1289/ehp.01109983
  9. Heikenwälder H, Heikenwälder M. Krebs - Lifestyle und Umweltfaktoren als Risiko. Bindemittel, Ballaststoffe und Darmentzündungen. Springer; 2019. P. 69-78.
  10. David S, Shani Levi C, Fahoum L, Ungar Y, Meyron-Holtz EG, Shpigelman A, et al. Revisiting the carrageenan controversy: do we really understand the digestive fate and safety of carrageenan in our foods? Food Funct. 2018;9(3):1344-1352. doi: 10.1039/c7fo01721a.
  11. Tkachenko AS, Onishchenko AI, Gorbach TV, Gubina-Vakulyсk GI. O-6-methylguanine-DNA methyltransferase (MGMT) overexpression in small intestinal mucosa in experimental carrageenan-induced enteritis. Malay. J. Biochem. Mol. Biol. 2018;21(3):77-80.
  12. Tkachenko A, Marakushyn D, Kalashnyk I, Korniyenko Y, Onishchenko A, Gorbach T, et al. A study of enterocyte membranes during activation of apoptotic processes in chronic carrageenan-induced gastroenterocolitis. Med Glas (Zenica). 2018;15(2):87-92. doi: 10.17392/946-18.
  13. Gubina-Vakyulyk GI, Gorbach TV, Tkachenko AS, Tkachenko MO. Damage and regeneration of small intestinal enterocytes under the influence of carrageenan induces chronic enteritis. Comp. Clin. Path. 2015;24(6):1473–1477. https://doi.org/10.1007/s00580-015-2102-3
  14.  Bhattacharyya S, Xue L, Devkota S, Chang E, Morris S, Tobacman JK. Carrageenan-induced colonic inflammation is reduced in Bcl10 null mice and increased in IL-10-deficient mice. Mediators Inflamm. 2013;2013(10):397642.
  15.  Bhattacharyya S, Gill R, Chen M-L, Zhang F, Linhardt RJ, Dudeja PK, et al. Toll-like receptor-4 mediates induction of Bcl10-NFκB-IL-8 inflammatory pathway by carrageenan in human intestinal epithelial cells. J Biol Chem. 2008;16(283):10550–8
  16.  Martino JV, Van Limbergen J, Cahill LE. The role of carrageenan and carboxymethylcellulose in the development of intestinal inflammation. Front Pediatr. 2017;5:96. doi: 10.3389/fped.2017.00096.
  17.  Biancone L, Armuzzi A, Scribano ML, Castiglione F, D'Incà R, Orlando A, et al. Cancer risk in inflammatory bowel disease: a 6-year prospective multicenter nested case-control IG-IBD study. Inflamm Bowel Dis. 2020;26(3):450-459. doi: 10.1093/ibd/izz155.
  18.  Rizzello F, Spisni E, Giovanardi E, Imbesi V, Salice M, Alvisi P, et al. Implications of the westernized diet in the onset and progression of IBD. Nutrients 2019;11:1033.
  19.  Bhattacharyya S, Shumard T, Xie H, Dodda A, Varady KA, Feferman L, et al. A randomized trial of the effects of the no-carrageenan diet on ulcerative colitis disease activity. Nutr Healthy Aging. 2017;4(2):181-192. doi: 10.3233/NHA-170023.
  20.  TkachenkoAS, Gubina-VakulyckGI, KlochkovVK, KavokNS, OnishchenkoAI, GorbachTV, et al. Experimental evaluation of the impact of gadolinium orthovanadate GdVO4:Eu3+ nanoparticles on the carrageenan-induced intestinal inflammation. Acta Medica (Hradec Králové). 2020;63(1):18–24 https://doi.org/10.14712/18059694.2020.11
  21.  Kopanytsia OM, Marushchak MI, Krynytska IY. Carrageenan induces cell death in rats blood. International Journal of Medicine and Medical Research. 2018;4(1):67-70. https://doi.org/10.11603/ijmmr.2413-6077.2018.1.8979
  22.  Tkachenko A. Reactive oxygen species (ROS) generation by lymphocytes in rats treated with a common food additive E407a. J Clin Med Kaz. 2020;1(55):22-26. DOI: 10.23950/1812-2892-JCMK-00744
  23.  Sokolova EV, Karetin Y, Davydova VN, Byankina AO, Kalitnik AA, Bogdanovich LN, et al. Carrageenans effect on neutrophils alone and in combination with LPS in vitro. J Biomed Mater Res A. 2016;104(7):1603-9. doi: 10.1002/jbm.a.35693.
  24.  Tkachenko AS, Onishchenko AI, LesovoyVN, Myasoedov VV. Common food additive E407a affects BCL-2 expression in lymphocytes in vitro.Studia Univ. VG, SSV, 2019;29(4):169-76.
  25.  McKim JM, Willoughby JA Sr, Blakemore WR, Weiner ML. Clarifying the confusion between poligeenan, degraded carrageenan, and carrageenan: A review of the chemistry, nomenclature, and in vivo toxicology by the oral route. Crit Rev Food Sci Nutr. 2019;59(19):3054-3073. doi: 10.1080/10408398.2018.1481822.
  26.  McKim JM Jr, Baas H, Rice GP, Willoughby JA Sr, Weiner ML, Blakemore W. Effects of carrageenan on cell permeability, cytotoxicity, and cytokine gene expression in human intestinal and hepatic cell lines. Food Chem Toxicol. 2016;96:1-10. doi: 10.1016/j.fct.2016.07.006.
  27.  Wu W, Zhen Z, Niu T, Zhu X, Gao Y, Yan J, et al. κ-Carrageenan enhances lipopolysaccharide-induced interleukin-8 secretion by stimulating the Bcl10-NF-κB Pathway in HT-29 cells and aggravates C. freundii-induced inflammation in mice. Mediators Inflamm. 2017;2017:8634865. doi: 10.1155/2017/8634865.
  28.  Ogata M, Matsui T, Kita T, Shigematsu A. Carrageenan primes leukocytes to enhance lipopolysaccharide-induced tumor necrosis factor alpha production. Infect Immun. 1999;67(7):3284–3289.