Research On Preventive Effects Of MaFGF On Rats' Renal Tubular Epithelial Cells Damaged By Gentamycin Sulfate In Vitro

Objective:To explore the preventive role of MaFGF in the injury of gentamicin on renal tubular epithelial cells of SD rats.Method:SD rat'primary renal tubular epithelial cells were sterilely colleccted, and then cultured in 6-well plates,96-well plates and culture bottles: (1) After the cells grow in logarithmic growth phase in 96-well plates, the cells were divided into five groups randomly. Control group were incubated for 36h normally; GM group were incubated with DMEM culture medium containing GM respectively and the final concentrations were followed by 1.0g/L,2.0 g/L,3.0 g/L,4.0 g/L and 5.0g/L at the latter 24 hours; MaFGF groups(Ⅰ,Ⅱ,Ⅲ) were incubated with DMEM culture medium containing MaFGF respectively and the final concentrations were followed by 5.20μg/L,6.24μg/L and 7.28μg/L at the former 12 hours, and then added GM as the above serious of drug concentrations at the latter 24 hours. Assay the activity of each group cells by MTT methord and calculate the inhibition rate and the corresponding IC50 values of GM. (2) The cells were divided into three groups randomly when they grow in logarithmic growth phase:Control group, GM group and MaFGF+GM group. Control group were incubated for 36h normally; GM group were incubated with DMEM culture medium containing 3.0g/L GM at the latter 24 hours; MaFGF+GM group were incubated with DMEM culture medium containing 7.28μg/L MaFGF at the former 12 hours, and then added 3.0 g/L GM at the latter 24 hours. And the preventive role of MaFGF in the injured renal epithelial cells was observed by using inverted phase contrast microscope and some biochemical and enzymological indexes.Result:(1) MaFGF pretreatment renal tubular epithelial cells,which can increase IC50 value of GM. (2) Added 2.0g/L or higher concentrations of GM at the latter 24 hours, renal tubular epithelial cells became abnormal shape, which lost the original multilateral cobblestone-like shape and became rounded shrinkage. Cell borders became fuzzier or even disappear.Cytoplasm became thin, and three dimensional sense became worse, and cell concentration was low. More black particles and vacuoles existed in cytoplasm. Some cells even were sheded and suspended in culture medium. But the cells which were pretreated whith MaFGF at the former 12 hours and added 3.0g/L GM at the latter 24 hours also showed that the extent of rounded shrinkage was remission, and the cell shapes became clearer, and the refraction of cell boundary was better. Cytoplasm became abundant, and three dimensional sense became better, and cell concentration was higher. A litter of black particles and vacuoles existed in cytoplasm. (3) Compared GM group and Control group, the content of MDA and NO was increased while the activity of SOD and GSH-Px was decreased, the differences were significant statistically (P<0.01); Compared MaFGF+GM group and GM group, the content of MDA and NO was decreased while the activity of SOD and GSH-Px was increased, the differences were significant statistically (P<0.01); Compared MaFGF+GM group and Control group, the content of the four indexes were not restored to the normal levels, the differences were still significant difference statistically (P<0.01).Conclusion:MaFGF could prevent the injury of renal tubular epithelial cells in some degree. The preventive effect of MaFGF on GM nephrotoxicity was related to antagonize free radicals directly or indirectly, enhance the activity of antioxidant enzymes which could scaven free radicals, and protect antioxidant enzymes.