Investigation of the effects of Inula viscosa L. in rainbow trout gonad cells (RTG-2) induced by lipopolysaccharide (LPS)

In this study, the anti-proliferative and anti-inflammatory effects of I. viscosa infected with LPS were investigated in gonad cell line isolated from rainbow trout (RTG-2). Preliminary studies were carried out the toxic dose determinations for the cells with different concentrations of LPS (1, 5, 10 and 20 µM/ml) and I. viscosa (IV) (1, 5, 10 and 20 µg/ml) were studied by viability tests. According to the obtained data, it was found that the concentration of 5 µM/ml LPS showed 2% proliferation compared to the control group, and decreases of 4% and 27% were recorded in 10 and 20% µM/ml, respectively. Therefore, 20 µM/ml LPS for experimental infection was selected as the active concentration. I. viscosa showed 1.1% cell proliferation with a concentration of 1µg/ml compared to the control group, reducing effects recorded with increasing effects (5, 10 and 20 µg/ml) on cell viability of 1.7%, 19% and 36%, respectively. The concentration of I. viscosa with anti-cytotoxic activity was 1 µg/ml. Then, four experimental instruments were designed. The first group was evaluated as a control group (C) the second group was infected with LPS (LPS). The third group treated with only LPS (LPS). The last group was formed by exposure of I. viscosa (LPS + IV) on gonad cells infected LPS. The essential essential oil of I. viscosa was provided by the method of water distillation. Expressions of IL-1β, IL-8 and TNF-α genes targeted for anti-inflammatory effect were determined by qRT-PCR method. ELISA method was used to detect IL-6 protein levels. All pro-inflammatory parameters investigated in the cells of RTG-2 LPS-infected cells up-regulated, increased TNF-α gene expression level (11.3 times). In the group with IV added at the end of LPS exposure, down regulation was monitored and the most effect was IL-1β with 96%. When the data for the IL-6 protein level were evaluated, the opposite was seen, it was determined that there was a 4% decrease by LPS compared to the control group and an increase of 9% in the group with IV addition at the end of LPS exposure. The efficacy of I. viscosa on pro-inflammatory cytokines was demonstrated by this study, and LPS inhibition and anti-inflammatory effects of IV against LPS were determined. However, further studies suggest that pharmacological activity of I. viscosa should be studied in more detail in many respects.