The Inhibitive Effect Of Mycophenolic Acid On Pterygium Fibroblasts

Objective:To investigate the inhibitive effect of MPA on PFB and relative mechanism of the effect.Methods:PFB and TFB were cultivated in vitro, identified by the method of vimentin immunohistochemisty. PFB and TFB in the third growing period were inoculated, and MTT was used to detect the proliferation ability at day1,4, and6. The method of MMT colorimetry and bromodeoxyuridine (BrdU) incorporation was used to investigate the effect of MPA at different concentration on PFB.4’,6-diamidino-2-phenylindole (DAPI) staining was used to show the morphology of TFB after co-cultivated with different concentration of MPA. Flow cytometry was used to detect apoptosis and periodicity cycle of PFB and TFB after co-cultured with different concentrations of MPA. MMT colorimetry was also used to show the difference between PFB and TFB, assess the security. P65and IkB-α in the pathway of NF-KB in PFB was detected by the method of immunohistochemisty and western blot.Results:PFB had stronger proliferation ability than TFB. Three days after cultivation, more numbers of PFB were found than TFB (p<0.05), a phenomenon which was more obvious in day4(p<0.01). MPA at all different concentrations had evident inhibitive effect on PFB.DAPI immunofluorescence staining found that there was no difference in nuclear morphology between different drug groups and control groups. Flow cytometer showed that MMC promote apoptosis or necrosis of TFB, which had direct proportion to concentration. However, the same effect was not found in MPA.There were no difference in inhibition ratio between PFB and TFB at72h after co-cultured with MPA. Conversely, under similar conditions, significant differences were found after PFB or TFB co-cultured with MMC. Mmunohistochemisty and western blot found that, MPA could inhibit the activation of NF-KB pathway, discharge of inflammatory and immune factors, finally proliferation of PFB by suppressing nuclear transformation.Conclusions: MPA can suppress proliferation of PFB without inducing apoptosis or necrosis of PFB, which may be related to its inhibitive effect on NF-KB of PFB.