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    Antioxidant effects of oleuropein on hydrogen peroxide-induced neuronal stress-an in vitro study
    (Bentham Science Publishers, 2020) Kucukgul, Altug; Isgor, Mehmet M.; Duzguner, Vesile; Atabay, Meryem N.; Kucukgul, Azime
    Background: Persistent oxidative stress can lead to chronic inflammation and mediate most chronic diseases including neurological disorders. Oleuropein has been shown to be a potent antioxidant molecule in olive oil leaf having antioxidative properties. Objective: The aim of this study was to investigate the protective effects of oleuropein against oxidative stress in human glioblastoma cells. Methods: Human glioblastoma cells (U87) were pretreated with oleuropein (OP) essential oil 10 µM. After 30 minutes, 100 µM H2 O2 was added to the cells for three hours. Cell survival was quantified by colorimetric MTT assay. Glutathione level, total oxidant capacity, total antioxidant capacity and nitric oxide levels were determined by using specific spectrophotometric methods. The relative gene expression level of iNOS was performed by qRT-PCR method. Results: According to viability results, the effective concentration of H2 O2 (100µM) significantly decreased cell viability and oleuropein pretreatment significantly prevented the cell losses. Oleuropein regenerated total antioxidant capacity and glutathione levels decreased by H2 O2 exposure. In addition, nitric oxide and total oxidant capacity levels were also decreased after administration of oleuropein in treated cells. Conclusion: Oleuropein was found to have potent antioxidative properties in human glioblastoma cells. However, further studies and validations are needed in order to understand the exact neuroprotective mechanism of oleuropein. © 2020 Bentham Science Publishers.
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    Apoptotic effects of artificial feed supplemented with Thymus vulgaris on Oncorhynchus mykiss against Yersinia ruckeri
    (Natl Inst Science Communication-Niscair, 2017) Kucukgul, Altug; Kucukgul, Azime
    Chemical composition of the essential oil was analyzed by GC-MS, and found to contain the phenol (40.95 %) as major component. Results showed that Y. ruckeri (3 x 10(7)cfit fish(-1)) infection caused the characteristic symptoms on rainbow trout (petechial hemorrhages on the surface of body and liver). Y. ruckeriinducedinhibitions of Cas-3, Cas-8, p53 and HSP-70. After the administration of thyme essential oil (10 mg kg(-1) diet) to infected fish tissues, Cas-3, Cas-8, p53 and HSP-70 mRNAs were up-regulated by 9.31, 9.18, 9.58, and 6,1 fold change, respectively. According to the results of the study, the inhibitions seen all analyzed genes were due to Y. ruckeri infection. However, they were entirely increased by thyme treatment. Thus, targeting proapoptotic proteases by agents such as T. vulgaris could improve therapeutic options for infectious disease in fish.
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    Investigation of the anti-apoptotic activity of ozone therapy in rainbow trout macrophages infected with Yersinia ruckeri
    (Springer, 2019) Kucukgul, Azime; Kucukgul, Altug; Gonenci, Ramazan; Ozsoy, Sule Yurdagul; Kutlu, Banu; Isgor, 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 24h after being incubated with the ozonized medium at nontoxic concentration. Effective concentrations of nontoxic ozone (3 gamma) and bacteria (1.5x10(8)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.
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    Investigation on the effects of Inula viscosa L. on rainbow trout gonad cells induced by lipopolysaccharide
    (Bdfish, 2021) Kucukgul, Azime; Kucukgul, Altug; Korkmaz, Tulay Metin
    Fish cells have been accepted as an alternative to in vivo assay for inflammatory effects of therapeutic experimental systems. To reveal the anti-inflammatory effects of Inula viscosa (IV) fish cell line RTG-2, derived from the gonadal tissue of rainbow trout, was infected with lipopolysaccharide (LPS). Effective concentrations for different concentrations of LPS (1, 5, 10, 20 mu M) and IN (1, 5, 10, 20 mu g ml(-1)) were determined. While the 20 mu M concentration of LPS, which was effectively selected from preliminary tests, caused 27% cell loss, the effective 1 mu g ml-1 concentration of IN caused 1.1% proliferation in the cells when compared to the control group. All pro-inflammatory parameters investigated in LPS-induced RTG-2 cells showed up-regulation, with the highest increase in TNF-alpha gene expression level (11.3 fold changes). Down-regulation was determined in the IN together with LPS administered group and IL-1 beta had the highest effect with 96%. IL-6 protein level decreased by LPS at a rate of 4% and IN together with LPS increased by 9%. The IN exhibited significant efficacy against inflammation caused by LPS. However, further studies are needed to determine pharmacological activity of I. viscosa in details.