Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
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Endotoxin of Porphyromonas gingivalis amplifies the inflammatory response in hyperglycemia-induced zebrafish through a mechanism involving chitinase-like protein YKL-40 analogs

  • Gizem Gunduz (Department of Periodontology, Institute of Health Sciences, Marmara University) ;
  • Merih Beler (Department of Biochemistry, Institute of Health Sciences, Marmara University) ;
  • Ismail Unal (Department of Biochemistry, Institute of Health Sciences, Marmara University) ;
  • Derya Cansiz (Department of Biochemistry, Faculty of Medicine, Istanbul Medipol University) ;
  • Ebru Emekli-Alturfan (Department of Biochemistry, Faculty of Dentistry, Marmara University) ;
  • Kemal Naci Kose (Department of Periodontology, Faculty of Dentistry, Marmara University, Marmara University Basibuyuk Medical Campus)
  • Received : 2023.01.12
  • Accepted : 2023.05.10
  • Published : 2023.10.15
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Abstract

Porphyromonas gingivalis (P. gingivalis), a key pathogen in periodontal diseases, is also associated with hyperglycemia-associated systemic diseases, including diabetes mellitus (DM). Gingipains are the most important endotoxins of P. gingivalis, and in vivo studies using gingipains are scarce. Zebrafish (Danio rerio) is a vertebrate with high physiological and genetic homology with humans that has multiple co-orthologs for human genes, including inflammation-related proteins. The aim of our study was to determine the effects of gingipain in a hyperglycemia-induced zebrafish model by evaluating inflammation, oxidant-antioxidant status, and the cholinergic system. Adult zebrafish were grouped into the control group (C), hyperglycemia-induced group subjected to 15 days of overfeeding (OF), gingipain-injected group (GP), and gingipain-injected hyperglycemic group (OF+GP). At the end of 15 days, an oral glucose tolerance test (OGTT) was performed, and fasting blood glucose (FBG) levels were measured. Lipid peroxidation (LPO), nitric oxide (NO), glutathione (GSH), glutathione S-transferase, catalase, acetylcholinesterase (AChE), alkaline phosphatase (ALP), and sialic acid (SA) levels were determined spectrophotometrically in the hepatopancreas. The expression levels of tnf-α, il-1β, ins, crp, and the acute phase protein YKL-40 analogs chia.5 and chia.6 were evaluated by RT-PCR. After two weeks of overfeeding, significantly increased weight gain, FBG, and OGTT confirmed that the zebrafish were hyperglycemic. Increased oxidative stress, inflammation, and AChE and ALP activities were observed in both the overfeeding and GP groups. Amplification of inflammation and oxidative stress was evident in the OF+GP group through increased expression of crp, il-1β, chia.5, and chia.6 and increased LPO and NO levels. Our results support the role of gingipains in the increased inflammatory response in hyperglycemia-associated diseases.

Keywords

Acknowledgement

The authors would like to thank Marmara University Scientific Research and Project Commission, Project No: TDK-2021-10420 for funding this project.

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