Regulation Of NF-Kappa B Inhibitor PDTC On The Uranium Mineral Dust Induced Macrophage Inflammation

Objective: To investigate the regulation effect of nuclear transcription factor-kappa B(NF-?B) on inflammation related active substances tumor necrosis factor-?(TNF-?), interleukin-6(IL-6), monocyte chemotactic protein-1(MCP-1), inducible nitric oxide synthase(i NOS) and nitric oxide(NO) in the uranium mineral dust induced rat macrophage RAW264.7 inflammatory reaction, and to analyze the role of NF- kappa B in the mechanism of uranium mineral dust induced lung injury for seeking the important target on the inhibition of uranium dust induced pulmonary fibrosis.Methods: The MTT method was used to detect the survival rate and time-efficiency of macrophage RAW264.7 exposed with different concentration of uranium dust for different time. The influence of expression of NF-kappa B p65 protein in nucleus and i NOS protein in cytoplasm induced by uranium mineral dust was test by Western Blot. The expression of TNF-?, IL-6, MCP-1, i NOS m RNA in macrophage was detected by Q-PCR. The expression of TNF-?, IL-6 and MCP-1 in cell culture supernatant was test by ELISA. And the expression of NO in cell culture supernatant was detected by nitrate reductase method.Results: The outcomes of MTT showed that after exposed to the uranium dust with the concentration of 0, 15, 30, 60, 120, 240?g/cm2 for 12, 24, 48 h, the survival rate of RAW264.7 presented a drop in a time-dose effective relationship. Treated with 60?g/cm2 uranium mineral dust, the expression of NF-kappa B p65 protein in nucleus presented a rising trend with the extension of time, and the expression of NF-kappa B p65 protein in uranium dust group was 5.10 times of the control group at 12h; The expression levels of TNF-?, IL-6, MCP-1 and i NOS m RNA in uranium dust group were higher than those in the control group; The content of TNF-?, IL-6 and MCP-1 in cell culture supernatant showed a upward trend with the extension of uranium dust treating time, and at 24 h their expression respectively reached(12827.78±518.77) pg/ml,(224.63±10.38) pg/ml,(4178.59±26.04) pg/ml,(202.30±10.62) ?mol/L in uranium dust group, which significantly higher than the control group; The expression of i NOS protein in cytoplasm showed a rising trend with the extension of uranium dust treating time, and the expression of i NOS protein in uranium dust group was 4.50 times of the control group at 24 h. After intervened with 25?mol / L PDTC, the expression levels of nucleus NF-kappa B p65 protein in PDTC group was close to the control group and significantly lower than that in uranium ore dust group; The content of TNF-?, IL-6 and NO in cell culture supernatant in PDTC group was significantly lower than that of uranium dust group and close to the control group; The content of MCP-1 in the supernatant showed no statistically significant differences between the PDTC group and the control group, and they were significantly higher than the control group; The expression levels of cytoplasm i NOS protein in PDTC group was close to the control group and significantly lower than that in uranium ore dust group.Conclusions:1.15-240?g/cm2 uranium dust can inhibit the cell proliferation of rat macrophage RAW264.7and induce the cell express NF-kappa B, TNF-?, IL-6, MCP-1 and i NOS and NO;2. 25?mol/L NF-kappa B inhibitor PDTC can inhibit macrophage RAW264.7 inflammatory injury induced by uranium dust in a certain extent.