Research On Protective Effects Of Afgf On Vascular Endothelial Cells Damaged By Cisplatin In Vitro

Objective: To explore the protective effects of acidic Fibrolast Growth Factor on the injury of cultured vascular endothelial cells induced by DDP.Method: The vascular endothelial cells of rats were obtained through tissue mass and identified by immunocytochemistry ofâ…§factor related antigen. After propagation and purification,the cells were inoculated in 96 well plates: The cells were divided into three groups randomly when they grow in logarithmic growth phase:Control group, DDP group and aFGF+DDP group. Main procedures are as follows:(1) After 24h cultivation,DDP with different concentration was added into the cultures,and MTT assay was performed to test the cell viability after 24h.(2) After 24h cultivation,aFGF with certain concentrations and DDP with different concentration were added into the cultures at the same time,and MTT assay was used to test the cell viability.(3)The protective effects of acidic Fibrolast Growth Factor on the injury of vascular endothelial cells induced by DDP was observed by using inverted phase contrast microscope and some biochemical and enzymological indexes changes such as MDA,NO,SOD and GSH-Px.Result: (1)The concentrations of DDP that inhibited 50% cell growth(IC₅₀) increased because of the effects of aFGF.(2) Added 2.59mg/L DDP afte 24h, vascular endothelial cells became abnormal shape, which lost the original single paving stones-like shape and became rounded shrinkage,three dimensional sense became worse,cells gap wider and connections reduced,cell borders became fuzzier or even disappeared,some cells even were sheded and suspended in culture medium.But when add 10μg/LaFGF and DDP together in it,after 24 hours,the cell morphology improvement,cells gap narrower and connections increased,the cell shapes became clearer,a small number of cells recovered for the short spindle.(3) Compared DDP group and control group, the content of MDA was increased while the activity of SOD,NO and GSH-Px were decreased,the differences were significant statistically (P<0.01);Compared aFGF + DDPgroup and DDP group, the content of MDA was decreased while the activity of SOD,NO and GSH-Px was increased,the differences were significant statistically (P<0.01);Compared aFGF +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: aFGF can exert certain protective effects on the vascular endothelial cells damaged by DDP.The protective effects of aFGF on vascular toxicity was caused by DDP related to they bind to cellular surface receptors,inhibit lipid over-oxidization,antagonize the damages of free radicals,antagonize free radicals directly or indirectly,and stabilize cellular enzyme activities to exert protective effects.