FIGURE 7.

NF-κB inhibitor(JSH23) reversed the up-regulation inflammation expression in Klotho knockdown cells and Klotho interacts with the NF-κB inhibitor IκBa in MH-S cells. A, klotho knockdown cells were treated with JSH23 for 24 h. The protein expressions of MMP-9 were detected by Western blotting. The mRNA expression of MMP-9 was detected by PCR. B, Klotho knockdown cells were treated with JSH23 for 24 h. The mRNA expression of TNF-α was detected by real time PCR. The release levels of TNF-α were determined by ELISA. C, klotho knockdown cells were treated with JSH23 for 24 h. The mRNA expression of IL-6 was detected by real time PCR. The release levels of IL-6 were detected by ELISA. D, Klotho was immunoprecipitated from MH-S cells treated with CSE (0.1%) for 1 h. Levels of IκBα were analyzed by immunoblotting. Then IκBα was immunoprecipitated from MH-S cells treated with CSE (0.1%) for 1 h. Levels of Klotho were analyzed by immunoblotting. CSE (0.1%) significantly decreased IκBα levels in the Klotho-IκBα complex. An equal amount (1 mg) of immunoprecipitated protein was used for Western blots. Data are the median of three independent experiments performed in triplicate. *, p < 0.05; **, p < 0.01, and ***, p < 0.001 compared with negative control siRNA group. #, p < 0.05; ##, p < 0.01, and ###, p < 0.001 compared with klotho siRNA group. Con, control.