Gizem GÜNDÜZ, Merih BELER, İsmail ÜNAL, Derya CANSIZ, Ebru EMEKLİ ALTURFAN, Kemal Naci KOSE

Gingipain Injection Affects Intestinal OxidantAntioxidant Status and Alkaline Phosphatase in Overfed Zebrafish

Objective: Porphyromonas gingivalis (P. gingivalis), a major periodontopathogen, is associated with overfeeding disorders, including metabolic syndrome. Gingipains are one of the most powerful endotoxins of P. gingivalis. Our aim was to reveal the effects of gingipain injections on the intestinal oxidant-antioxidant status and alkaline phosphatase (ALP) activity in overfed zebrafish. Materials and Methods: Four groups of healthy adult zebrafish were placed in random tanks as C: Control (n=15); GP: Gingipain (n=15); OF: Overfeeding (n=15); and OF+GP: Overfeeding+Gingipain (n=15) groups. At the end of the experiment, levels of intestinal lipid peroxidation (LPO) and ALP, glutathione S-transferase (GST), and catalase (CAT) activities were evaluated. Results: Intestinal LPO was significantly lower in the GP and OF groups compared to C. Gingipain injection in OF (OF+GP) significantly elevated LPO when compared to C, GP, and OF groups. ALP activities decreased significantly in the GP, OF, and OF+GP compared to C. GST activities increased significantly in the GP when compared to C. Decreased GST activities were observed in the OF and OF+GP. This decrease was less in OF+GP. CAT activities significantly decreased in all groups when compared to C. Conclusion: Our findings demonstrate that gingipain injection alters the ALP activity and intestinal oxidant-antioxidant status in overfed zebrafish.

Keywords:

Gingipain, zebrafish, oxidative stress,

PDF

___

1. Mysak J, Podzimek S, Sommerova P, Lyuya-Mi Y, Bartova J, Janatova T, et al. Porphyromonas gingivalis: Major periodontopathic pathogen overview. J Immunol Res 2014; 2014: 476068. google scholar
2. Xu W, Zhou W, Wang H, Liang S. Roles of Porphyromonas gingivalis and its virulence factors in periodontitis. Adv Protein Chem Struct Biol 2020; 120: 45-84. google scholar
3. Nunes JM, Fillis T, Page MJ, Venter C, Lancry O, Kell DB, et al. Gingipain R1 and lipopolysaccharide from Porphyromonas gingivalis have major effects on blood clot morphology and mechanics. Front Immunol 2020; 11: 1551. google scholar
4. Dominy SS, Lynch C, Ermini F, Benedyk M, Marczyk A, Konradi A, et al. Porphyromonas gingivalis in Alzheimer’s disease brains: Evidence for disease causation and treatment with small-molecule inhibitors. Sci Adv 2019; 5(1): eaau3333. google scholar
5. Tsuzuno T, Takahashi N, Yamada-Hara M, Yokoji-Takeuchi M, Sulijaya B, Aoki-Nonaka Y, et al. Ingestion of Porphyromonas gingivalis exacerbates colitis via intestinal epithelial barrier disruption in mice. J Periodontal Res 2021; 56(2): 275-88. google scholar
6. Mei F, Xie M, Huang X, Long Y, Lu X, Wang X, et al. Porphyromonas gingivalis and its systemic impact: Current status. Pathogens 2020; 9(11): 944. google scholar
7. Bullon P, Morillo J, Ramirez-Tortosa MC, Quiles J, Newman H, Battino M. Metabolic syndrome and periodontitis: Is oxidative stress a common link? J Dent Res 2009; 88(6): 503-18. google scholar
8. Nibali L, Tatarakis N, Needleman I, Tu Y-K, D’Aiuto F, Rizzo M, et al. Association between metabolic syndrome and periodontitis: A systematic review and meta-analysis. J Clin Endocrinol Metab 2013; 98(3): 913-20. google scholar
9. Dong Z, Lv W, Zhang C, Chen S. Correlation analysis of gut microbiota and serum metabolome with porphyromonas gingivalis-induced metabolic disorders. Front Cell Infect Microbiol 2022; 12: 858902. google scholar
10. Molnar K, Vannay Â, Szebeni B, Banki NF, Sziksz E, Cseh Â, et al. Intestinal alkaline phosphatase in the colonic mucosa of children with inflammatory bowel disease. World J Gastroenterol 2012; 18(25): 3254. google scholar
11. Panjamurthy K, Manoharan S, Ramachandran CR. Lipid peroxidation and antioxidant status in patients with periodontitis. Cell Mol Biol Lett 2005; 10(2): 255-64. google scholar
12. Karaman GE, Emekli-Alturfan E, Akyüz S. Zebrafish; an emerging model organism for studying toxicity and biocompatibility of dental materials. Cell Mol Biol (Noisy-le-grand) 2020; 66(8): 41-6. google scholar
13. Oka T, Nishimura Y, Zang L, Hirano M, Shimada Y, Wang Z, et al. Diet-induced obesity in zebrafish shares common pathophysiological pathways with mammalian obesity. BMC physiol 2010; 10(1): 1-13. google scholar
14. Dandin E, Üstündağ ÜV, Ünal İ, Ateş-Kalkan PS, Cansız D, Beler M, et al. Stevioside ameliorates hyperglycemia and glucose intolerance, in a diet-induced obese zebrafish model, through epigenetic, oxidative stress and inflammatory regulation. Obes Res Clin Pract 2022; 16(1): 23-9. google scholar
15. Zang L, Shimada Y, Nishimura Y, Tanaka T, Nishimura N. A novel, reliable method for repeated blood collection from aquarium fish. Zebrafish 2013; 10(3): 425-32. google scholar
16. Widziolek M, Prajsnar TK, Tazzyman S, Stafford GP, Potempa J, Murdoch C. Zebrafish as a new model to study effects of periodontal pathogens on cardiovascular diseases. Sci Rep 2016; 6(1): 36023. google scholar
17. Farrugia C, Stafford GP, Potempa J, Wilkinson RN, Chen Y, Murdoch C, et al. Mechanisms of vascular damage by systemic dissemination of the oral pathogen Porphyromonas gingivalis. FEBS J 2021; 288(5): 1479-95. google scholar
18. Wilensky A, Potempa J, Houri-Haddad Y, Shapira L. Vaccination with recombinant RgpA peptide protects against Porphyromonas gingivalis-induced bone loss. J Periodontal Res 2017; 52(2): 285-91. google scholar
19. Kose K, Gündüz G, Beler M, Emekli-Alturfan E. Inflammatory gene expression in response to gingipain in zebrafish. The 26th International Congress of Turkish Dental Association, September 8-11, J Clin Sci (Supplements), 2022; 11(3): 596-597. google scholar
20. Lowry OH. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193: 265-75. google scholar
21. Yagi K. Assay for blood plasma or serum. Methods Enzymol 1984; 105: 328-31. google scholar
22. Aebi H. Catalase in vitro. Methods Enzymol 1984; 105: 121-6. google scholar
23. Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases: The first enzymatic step in mercapturic acid formation. J Biol Chem 1974; 249(22): 7130-9. google scholar
24. Walter K, Schütt C. Acid and alkaline phosphatase in serum (two point method). Methods of Enzymatic Analysis. Vol. 2. Bergmeyer HU editor, Boca Raton, FL, USA: Verlag Chemie GmbH; 1974. p. 856-60. google scholar
25. Maloy KJ, Powrie F. Intestinal homeostasis and its breakdown in inflammatory bowel disease. Nature 2011; 474(7351): 298-306. google scholar
26. Tsuzuno T, Takahashi N, Yamada-Hara M, Yokoji-Takeuchi M, Sulijaya B, Aoki-Nonaka Y, et al. Ingestion of Porphyromonas gingivalis exacerbates colitis via intestinal epithelial barrier disruption in mice. J Periodontal Res 2021; 56(2): 275-88. google scholar
27. Estaki M, DeCoffe D, Gibson DL. Interplay between intestinal alkaline phosphatase, diet, gut microbes and immunity. World J Gastroenterol 2014; 20(42): 15650. google scholar
28. Röth E, Marczin N, Balatonyi B, Ghosh S, Kovacs V, Alotti N, et al. Effect of a glutathione S-transferase inhibitor on oxidative stress and ischemia-reperfusion-induced apoptotic signalling of cultured cardiomyocytes. Exp Clin Cardiol 2011; 16(3):92-6. google scholar
29. Frohnert BI, Bernlohr DA. Glutathionylated products of lipid peroxidation: a novel mechanism of adipocyte to macrophage signaling. Adipocyte 2014; 3(3): 224-9. google scholar