The effect of hydro alcoholic extract of Nigella sativa seeds on inflammatory mediators in C6 glioma cell line

Abstract

Nigella sativa (N. sativa) is a highly valued nutritional plant, which has long been used in traditional medicine to treat a variety of human diseases. The multifaceted pharmacological impacts of N. sativa, such as attenuating oxidative stress and inflammation, make it a suitable therapeutic candidate against cardiovascular, hepatic, and neurological disorders as well as cancer. Therefore, the current study aimed to evaluate the effect of the hydroalcoholic extract of N.sativa seeds on several pro-inflammatory cytokines in the C6 glioma cell line and to compare it with the effect of the extract on the normal fibroblast cell line. C6 and fibroblast cell lines were treated with the extract of N.sativa seeds, and 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay was performed to determine the half-maximal inhibitory concentration (IC50) after 72h of treatment. Real-time polymerase chain reaction (RT-PCR) was carried out to assess the expression levels of interleukin (IL)-6, IL-10, tumor necrosis factor-alpha (TNF-α), and transforming growth factor- β1 (TGF-β1) at the mRNA level in both cell lines after 72h of treatment with non-toxic and IC50 concentrations obtained from C6 cell line. The IC50 values for the hydroalcoholic extract of N.sativa seeds were 260±20μg/mL in the C6 cell line and 398±27μg/mL in fibroblast cells. The real-time PCR results indicated that the treatment of C6 and fibroblast cells with the extract at the IC50 value of N.sativa in C6 for 72h could increase the mRNA expression levels of IL-10 and reduce the mRNA expression levels of IL-6, TNF-α, and TGF-β1 in C6 and fibroblast cells. The N.sativa extract showed a higher anti-inflammatory effect on C6 cells in comparison with fibroblast cells. Regarding the anti-inflammatory effect of Nigella sativa in C6 cell line, it may be considered a promising candidate to fortify antitumor actions in combination with other therapeutic options in the treatment of patients with GBM.