Gündüz Gizem, Beler Merih, Ünal İsmail, Cansız Derya, Emekli-Alturfan Ebru, Kose Kemal Naci
Department of Periodontology, Institute of Health Sciences, Marmara University, Istanbul, Turkey.
Department of Biochemistry, Institute of Health Sciences, Marmara University, Istanbul, Turkey.
Toxicol Res. 2023 May 31;39(4):625-636. doi: 10.1007/s43188-023-00190-4. eCollection 2023 Oct.
(), 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 and in vivo studies using gingipains are scarce. Zebrafish () 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 and the acute phase protein YKL-40 analogs and 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, , and and increased LPO and NO levels. Our results support the role of gingipains in the increased inflammatory response in hyperglycemia-associated diseases.
牙龈卟啉单胞菌（Porphyromonas gingivalis）是牙周疾病的关键病原体，还与包括糖尿病（DM）在内的高血糖相关全身性疾病有关。牙龈蛋白酶是牙龈卟啉单胞菌最重要的内毒素，而使用牙龈蛋白酶的体内研究很少。斑马鱼（Danio rerio）是一种与人类具有高度生理和遗传同源性的脊椎动物，其具有人类基因的多个共同直系同源物，包括炎症相关蛋白。我们研究的目的是通过评估炎症、氧化-抗氧化状态和胆碱能系统，确定牙龈蛋白酶在高血糖诱导的斑马鱼模型中的作用。成年斑马鱼被分为对照组（C）、经过15天过度喂养（OF）的高血糖诱导组、牙龈蛋白酶注射组（GP）和牙龈蛋白酶注射高血糖组（OF+GP）。在15天结束时，进行口服葡萄糖耐量试验（OGTT），并测量空腹血糖（FBG）水平。采用分光光度法测定肝胰腺中的脂质过氧化（LPO）、一氧化氮（NO）、谷胱甘肽（GSH）、谷胱甘肽S-转移酶、过氧化氢酶、乙酰胆碱酯酶（AChE）、碱性磷酸酶（ALP）和唾液酸（SA）水平。通过RT-PCR评估crp、急性期蛋白YKL-40类似物ykl-40a和ykl-40b的表达水平。经过两周的过度喂养后，体重增加、FBG和OGTT显著增加，证实斑马鱼出现了高血糖。在过度喂养组和GP组中均观察到氧化应激、炎症以及AChE和ALP活性增加。通过crp、ykl-40a、ykl-40b的表达增加以及LPO和NO水平升高，OF+GP组中炎症和氧化应激的放大明显。我们的结果支持牙龈蛋白酶在高血糖相关疾病中炎症反应增加中的作用。
