Inhibition of TNF-αα/IFN-γγ induced RANTES expression in HaCaT cell by naringin

Objective: The effect of naringin on production of RANTES (regulated upon activation normal T-cell expressed and secreted) in human HaCaT cells was investigated here for the first time. Materials and methods: The HaCaT cells were cultured in Dulbecco''s modified Eagle''s medium (DMEM) and the proliferation of cell was determined by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT). The cells were divided into three groups including control group, tumor necrosis factor alpha (TNF-alpha alpha)/interferon gamma (IFN-gamma gamma)-stimulated group, and naringin pretreatment group (first incubated in the presence of naringin and then exposed to TNF-alpha alpha/IFN-gamma gamma). The concentration of RANTES in the supernatants was determined by enzyme-linked immunosorbent assay (ELISA). The expression of RANTES mRNA was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The expression of nuclear factor kappa B (NF-kappa kappa B) P65 protein was detected with immunocytochemical method and western blot method. Results: Naringin hardly inhibits HaCaT cells growth at concentrations rising from 0.25 to 1 mmol/L. However, RANTES expression detected in supernatant stimulated with TNF-alpha alpha/IFN-gamma gamma reduced 15 and 16%, respectively, when cultured with 0.25, 0.5 mmol/L naringin. Furthermore, 1 mmol/L naringin significantly decreased RANTES mRNA level. Finally, naringin decreased the expression of NF-kappa kappa B P65 protein in nuclei. Discussion and conclusion: Naringin can inhibit the increased production of RANTES, which is partially via NF-kappa kappa B-dependent signal pathway.