The Impact Of Human Amniotic Homogenate Supernatant On BFGF Expression In Rabbit Corneal Epithelia Cells

Objective: To investigate the expression level of rabbit corneal epithelial cells' bFGF ( basic fibroblast growth factor) in vitro under the condition of using different concentration of Human amniotic homogenate supernatant, and to discuss the function of human amniotic homogenate supernatant in the process of corneal epithelial repair.Methods: The rabbit corneal epithelial cells were cultured in vitro and they were divided into several groups. Then the experiment group was treated in DMEM culture liquid of 10% fetal blood serum mixed with different concentration of human amniotic homogenate supernatant (the amniotic homogenate protein concentration is 40μg/ml, 80μg/ml, and 160μg/ml respectively), with the control group treated in DMEM culture liquid of 10% fetal blood serum only. By using the method of MTT colorimetry, we aim to investigate the impact of human amniotic homogenate supernatant on rabbit corneal epithelial cells proliferation at different times (24h,48h,96h). At the same time, we tested the bFGF-mRNA expression level in each group by RT-PCR.Results:1. Rabbit corneal epithelial cells can proliferate at different concentrations of human amniotic homogenate supernatant (40μg/ml, 80μg/ml, 160μg/ml). The amount of corneal epithelial cells in each experimental group was increased significantly ( 0.185±0.010 , 0.318±0.015, 0.501±0.014 respectively, P<0.05) .As the culture time prolongs, the difference will be more apparent.2. There is no significant difference about the expression of bFGF-mRNA between the control (0.708±0.013) and the amniotic homogenate supernatant with the concertration of 40μg/ml and 80μg/ml (0.750±0.007, 0.785±0.006 respectively). But the differences about the expression of bFGF- mRNA between the 160μg /ml group (1.013±0.120) and the 40μg /ml, 80μg /ml, control group are significant respectively.Conclusion:1. Human amniotic homogenate supernatant can significantly stimulate the proliferation of rabbit corneal epithelial cells and its effect shows dose-dependant.2. There is the expression of bFGF-mRNA in rabbit corneal epithelial cells that were cultured in vitro.3. Human amniotic homogenate supernatant could promote the expression of the bFGF-mRNA in rabbit corneal epithelium cells in vitro, which is concentration related.