The Protective Effect of Nobiletin on Lipopolysaccharides (LPS)-induced RAW 264.7 Cell Injury
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Abstract:
To investigate the protective effects of nobiletin on LPS-induced RAW 264.7 macrophage cell injury. The MTT method was used to detect the safe dosage of nobiletin. The release level of lactate dehydrogenase (LDH) was measured by the LDH assay kit. The level of nitric oxide (NO) released in the cell supernatant was evaluated by the NO assay kit. The mRNA expression levels of TNF-α, IL-1β, iNOS, TLR4 and CD14 were evaluated by real-time PCR analysis. The expression level of NF-κB nuclear protein was determined by the western blot method. Compared to the normal control group, the LDH activity and amount of released NO in the LPS-stimulated group increased by 4.12 times and 3.21 times, respectively (p<0.01), and the mRNA expression levels of iNOS, IL-1β, TNF-α, TLR4 and CD14 increased by 2.73 times, 4.91 times, 10 times, 1.84 times and 3.53 times, respectively (p<0.01). Meanwhile, the nuclear expression level of NF-κB in the LPS-stimulated group increased by 2.50 times (p<0.01). However, the treatment with nobiletin at different doses decreased the activity of LDH and amount of the released NO, while reducing the mRNA expression levels of TNF-α, IL-1β, iNOS, TLR4 and CD14 as well as the nuclear expression level of NF-κB, compared with the LPS-stimulated group (p<0.05 or p<0.01). Nobiletin attenuates LPS-induced injury in RAW 264.7 cells through mechanisms probably associated with the inhibition of TLR4-NF-κB signaling pathway.
