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Protective Effect of Modified Human Acidic Fibroblast Growth Factor against Actinomycin D-Induced NRK52E Cells Apoptotic Death
Abstract
Purpose: To investigate whether modified acidic fibroblast growth factor (MaFGF) can protect NRK52E cell against apoptotic death induced by actinomycin D (Act D) and the effect of MaFGF on PI3K/Akt signaling pathway.
Methods: NRK52E cell apoptotic death was measured by several methods including cell morphologic observation, Hoechst 33342 staining and flow cytometry. In addition, the levels of phosphorylated-Akt protein were analyzed by Western blotting method.
Results: The results showed that 0.75 mg/L Act D-treated NRK52E cell for 20 h was the optimal conditions for establishing NRK52E cell apoptotic model. Different doses of MaFGF (0.01, 0.03, 0.1, 0.3 and 1.0 mg/L) decreased apoptotic rate but enhanced the expression of phosphorylated Akt protein. However, MaFGF’s protection against Act D-induced apoptosis was significantly (p < 0.05) prevented when NRK52E cells were exposed to wortmannin.
Conclusion: These results reveal that MaFGF can reduce the level of ActD-induced apoptotic cell death in 20 h, and the protective mechanism of MaFGF may be associated with the activation of PI3K/Akt signaling pathway by up-regulation of expression of phosphorylated Akt protein.
Keywords: Modified acidic fibroblast growth factor (MaFGF), Renal injury, Apoptotic death, Actinomycin D (Act D)