The Effect Of Tripterine On Proliferation And Apoptosis Of Mouse Retinal Endothelial Cells

Objective :The purpose of this study was to investigate the effect of tripterine on proliferation and apoptosis and expression of vascular endothelial growth factor of mouse retinal endothelial cells,and to provide a new clue and theory basis for developing a drug of preventing and curing retinal neovascularization in the future.Methods : MREC were primary cultured, and identified by the positive expression of FⅧr-Ag using immunocytochemistry. The cultured MREC were treated with different concentration of tripterine , the effect of cultured MREC proliferation was measured by MTT, the effect of cultured MREC apoptosis was measured by flow cytometer. The expression of VEGF was identified by using immunocytochemistry; the positive control group used angiostatin (150μg/ml).Results :1.Immunohistochemistry techniques showed that FⅧr-Ag was the positive expression in MREC;2. MTT assay indicated that tripterine significantly suppressed proliferation of MREC in dose-dependant and time-dependant. The inhibitory rate were 8.62±4.23%,24.41±6.27%,46.51±5.95%,65.62±9.21%,65.62±9.21%,74.69±11.26% after they were incubated with tripterine for 24 hours, they are 9.34±3.78%,26.21±7.35%, 51.36±6.39%,66.59±8.99%,73.19±10.24%,76.16±15.21% for 48 hours and 11.24±4.62%, 32.69±7.21%,55.28±10.25%,70.22±9.34%,75.84±13.69%,78.93±10.76%.for 72 hours.3. MREC were treated with different concentration of tripterine(0.01μg/ml,0.02μg/ml, 0.05μg/ml,0.1μg/ml)on 24 hours,PI staining flow cytometry showed that the apoptosis rate of MREC was 8.42±1.07% with tripterine concentration of 0.05μg/ml. It was different with control group disposed with DMEM (P<0.05),Compared with the An group, It was not different (P>0.05);4.The different concentration of tripterine (0.01μg/ml,0.02μg/ml,0.05μg/ml,0.1μg/ml) treated to MREC on 24 hours, Immunohisto- chemical techniques indicated that the expression of VEGF in MREC endochylema was decreased with increasing the concentration of tripterine(P<0.05) Conclusions :1. tripterine can effectively inhibit the proliferation of MREC, the inhibitory rates are increased in both time-dependent and dose -dependent ways in vitro. 2. tripterine can effectively induce the apoptosis of MREC in vitro. 3. Immunohistochemistry techniques showed that the effects of growth inhibition of tripterine on MREC are associated with down-regulating expression of VEGF protein.