Publication: Evaluation of the effects of porphyromonas gingivalis gingipain on zebrafish embryo
Abstract
Amaç: Periodontitisin oluşumunda rol oynayan esas patojenlerden olan Porphyromonas gingivalis, olumsuz gebelik sonuçları da dahil çok sayıda sistemik hastalık ile ilişkilendirilmektedir. Bu çalışmada, gingipainin zebra balığı embriyoları üzerindeki etkisinin, embriyotoksisite biyobelirteçleri olarak kabul edilen bazı genlerin (gfap, erg, igf2 ve gclc) ekspresyonlarındaki ve oksidan-antioksidan sistemi üzerindeki etkilerinin incelenmesi amaçlandı. Gereç ve Yöntem: Zebra balığı embriyoları 72 hpf'ye kadar 100 ng/ mL ve 200 ng/ mL konsantrasyonlarda gingipain RgpA'ya maruz bırakıldıktan sonra homojenize edildi. Gingipainin, büyüme/ gelişim ile ilişkili bazı genlerin (erg, igf2, gclc ve gfap) ekspresyonları üzerindeki etkisi RT-PCR yöntemi ile değerlendirildi. Oksidan-antioksidan sistem komponentlerinin düzeyleri spektrofotometrik yöntemlerle belirlendi. Bulgular: Gingipaine maruz bırakılan embriyolarda morfolojik malformasyonlar gözlemlendi. Her iki gingipain dozuna maruz kalan gruplarda nitrik oksit seviyelerinde artış, glutatyon-S-transferaz aktivitesinde ve süperoksit dismutaz aktivitelerinde azalma gözlendi. gfap, gclc, erg genlerinin ekspresyonlarında değişiklik gözlemlenirken, igf-2'nin ekspresyonunda farklılık görülmedi. Sonuç: Bu çalışmanın sonuçları, P. gingivalisin en önemli endotoksinlardan olan gingipainin, zebra balığı embryoları üzerinde toksik etkiye sahip olduğu ve gelişimsel bozukluklara neden olabileceği gösterilmiştir. Bu bulgular, gingipainler ile olumsuz gebelik sonuçları arasındaki olası ilişkiyi göstermekte olup bu konuda yapılacak daha ileri çalışmaların önünü açacaktır.
Objective: Porphyromonas gingivalis, which is the main pathogen involved in the onset and progression of periodontitis, has long been associated with numerous systemic diseases including adverse pregnancy outcomes. In this study, it was aimed to investigate the effects of gingipain on zebrafish embryos, which have become a popular vertebrate model in recent years, and its effects on certain genes recognized as embryotoxicity biomarkers, gfap, erg, igf2 and gclc, as well as the oxidant-antioxidant system. Materials and Methods: Zebrafish embryos were exposed to RgpA gingipain at 100 ng/ mL and 200 ng/ mL concentration up to 72 hpf and then homogenized. The effects on expression of some developmental genes (erg, igf2, gclc, and gfap) were assessed using RT-PCR methods, and the levels of oxidant and antioxidant system components were determined by spectrophotometric methods. Results: Morphologic malformations were observed in zebrafish embryos exposed to gingipain. Not only an increase in nitric oxide and lipid peroxidation levels, in addition a decrease in glutathione-S-transferase activity and superoxide dismutase activities were observed in both gingipain exposed groups. The expressions of gfap, gclc, erg genes were dysregulated while that of igf-2 did not show any significant change. Conclusion: Our study demonstrated the embryotoxicity nature of gingipains, which is the strongest endotoxin of P. gingivalis, and that exposure to gingipain could lead to developmental disturbances in the embryonic period in zebrafish. These findings indicate the possible relationship between gingipains and adverse pregnancy outcomes as a subject worth investigating.
Objective: Porphyromonas gingivalis, which is the main pathogen involved in the onset and progression of periodontitis, has long been associated with numerous systemic diseases including adverse pregnancy outcomes. In this study, it was aimed to investigate the effects of gingipain on zebrafish embryos, which have become a popular vertebrate model in recent years, and its effects on certain genes recognized as embryotoxicity biomarkers, gfap, erg, igf2 and gclc, as well as the oxidant-antioxidant system. Materials and Methods: Zebrafish embryos were exposed to RgpA gingipain at 100 ng/ mL and 200 ng/ mL concentration up to 72 hpf and then homogenized. The effects on expression of some developmental genes (erg, igf2, gclc, and gfap) were assessed using RT-PCR methods, and the levels of oxidant and antioxidant system components were determined by spectrophotometric methods. Results: Morphologic malformations were observed in zebrafish embryos exposed to gingipain. Not only an increase in nitric oxide and lipid peroxidation levels, in addition a decrease in glutathione-S-transferase activity and superoxide dismutase activities were observed in both gingipain exposed groups. The expressions of gfap, gclc, erg genes were dysregulated while that of igf-2 did not show any significant change. Conclusion: Our study demonstrated the embryotoxicity nature of gingipains, which is the strongest endotoxin of P. gingivalis, and that exposure to gingipain could lead to developmental disturbances in the embryonic period in zebrafish. These findings indicate the possible relationship between gingipains and adverse pregnancy outcomes as a subject worth investigating.