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- MakaleYalnız künye2019Scopus SJRQ2Investigation of the anti-apoptotic activity of ozone therapy in rainbow trout macrophages infected with Yersinia ruckeri
Küçükgül, Azime; Küçükgül, Altuğ; Gönenci, Ramazan; Özsoy, Şule Yurdagül; Kutlu, Banu; Işgör, Mehmet Mustafa
Yersinia ruckeri is the causal agent of enteric redmouth (ERM) in trout. This study aimed to investigate the effectiveness of nontoxic ozone on Y. ruckeri-infected trout cells in in vitro conditions. Rainbow trout macrophages (RTS-11) were exposed to Y. ruckeri for 24 h after being incubated with the ozonized medium at nontoxic concentration. Effective concentrations of nontoxic ozone (3 gamma) and bacteria (1.5 x 108 cfu/mL) were determined by viability tests. The expression of caspases 1, 3, 8, and 12 was determined by real-time PCR and protein levels investigated by ELISA method. Apoptotic-necrotic cell ratios were determined by acridine orange and ethidium bromide staining. Bacterial stimulations of caspases (1, 3, 8, and 12) were suppressed by ozone mostly at the translational level. While bacterial infection increased the count of apoptotic and necrotic cells at different stages, ozone significantly reduced this condition. Obtained data indicate possible use of nontoxic ozone as a safe and effective treatment alternative for increasing host defenses and eliminating the infection in the case of yersiniosis in fish.
AtıfWoS1OpenAlex3
İncele - Proje2017Yersinia ruckeri ile Enfekte Gökkuşağı Alabalığı Makrofajlarında Ozonoterapinin Antiapoptotik Etkinliğinin Araştırılması
Otay, Tuba; Küçükgül, Altuğ; Küçükgül, Azime; Özsoy, Şule Yurdagül; Kutlu, Banu; Gonenci, Ramazan
Yersiniosis is a systemic disease caused by Yersinia ruckeri, which is particularly affecting Enteric red mite (ERM) disease in trout. There are still many issues regarding the mechanism of apoptosis and necrosis that play a role in the pathogen-host relationship in the disease. In the project, it was aimed to maintain the life of the cells and killing the bacteria by providing anti-apoptotic and anti-necrotic effect via application of therapeutic concentration of ozone to the macrophages infected with Y. ruckeri. In this study, the cells of the rainbow trout-originated cell line (RTS-11, mononuclear / macrophage) were exposed to the appropriate number of bacteria (Yersinia ruckeri, ATCC- 29473, serotype 1) for 24 hours after being incubated with ozonized medium (30 min) at the nontoxic concentration. For this purpose, effective concentrations of nontoxic ozone and bacteria were demonstrated by viability tests (3 Gamma, 1.5x108, respectively). After incubation times, the expression levels of caspases 1,3,8 and 12 were determined by realtime polymerase chain reaction and protein levels were investigated by ELISA method from the obtained lysates. In addition, apoptotic-necrotic cell ratios were determined in acridine orange and ethidium bromide staining in experimental groups. According to the findings, bacterial infection caused 47% loss of cells, while nontoxic ozone was found to protect the cells by 27%. Again, while bacterial infection increased apoptotic cells in different processes, ozone was found to significantly reduce this rate. At the same time, ozone suppressed the rate of necrosis caused by bacteria. It was also found that bacterial stimulation of caspases (1, 3, 8 and 12), which were effective in the initiation of apoptosis and necrotic processes and in the formation of cell loss, were suppressed by ozone pre-administration and mostly at the level of protein translation. According to the obtained data, Yersinia ruckeri infection caused cell loss in the macrophages of rainbow trout by stimulating both apoptosis and necrosis. However, nontoxic ozone significantly improved this negative situation. In this context, it has come to the conclusion that nontoxic ozone can be used as a safe and effective treatment alternative for increasing host defenses and eliminating the infection in the case of yersiniosis in fishes. However, in order for this argument to be fully understood, it is inevitable to support it with advanced molecular and in vivo studies.
