Effect Of Berberine On The Proliferation And VEGF Expression Of Retinal Vascμlar Endothelial Cells In High Glucose Environment

Objective: To study the differenteffect of Berberine(BBR)on the cell proliferation of Human retinal vascular endothelial cells(HRCECs)and the expression of vascular endothelial growth factor(VEGF)under high glucose condition in vitro.Methods: Retinal vascμLar endothelial cells derived from the human eyeball were used in the experiment.Human retinal vascμLar endothelial cell model was established by stimμLating diabetic environment in high glucose medium,and the retinal vascμLar endothelial cells were identified by immunofluorescence.The experiment was divided into six groups as following: low glucose group,high glucose group,high glucose +5μmol/L berberine group,high glucose +10μmol/L berberine group,high glucose +20μmol/L berberine group,high glucose +40μmol/L berberine group.After the treatment of different concentrations of Berberine with HRCECs,,the effect of it on the cell proliferation of HRCECS in the high-glucose environment was detected by MTT method at 12,24 and 36 hours after exposure,and the expression of VEGF was also measured by western blot.ResμLts :1.MTT assay showed that:(1)the number of retinal vascμLar endothelial cells in the low-glucose group was significantly lower than that in the high-glucose group after 12,24 and 36 h incubation with different concentration of Berberine in vitro(P<0.05),.(2)when HRCEs were treated with berberine for the same time in the high-glucose environment,the cell numbers of HRCEs showed a significant decrease with the increase of concentrations of berberine(from5μmol/L to 40umoL),which showed an obvious dose-dependent cytotoxicity(P<0.05).(3)when HRCEs were incubated with berberine at the same concentration,we found with the long duration of exposure,the inhibitory effect of Berberine on cell proliferation showed a significant increase gradually,which showed a time-dependent cytotoxicity.(P<0.05).(4)compared with HRCECs in the high-glucose group,the higher the concentration of berberine and the longer the action time,the more obvious the inhibitory effect on cells was.2.Western blot showed that :(1)when HRCECs were co-cμLtured with Berberine for 12,24 and 36 h in vitro,the expression of VEGF in the high glucose group were significantly increased than those in the low glucose group(P<0.05).(2)when different concentrations berberine(5μmol/L,10μmol/L,20 umol,40umoL)were exposed to HRCECS for 12,24,and 36 h,respectively,the level of VEGF in cells showed a higher,which showed a significant dose-and time-dependent manner(P<0.05).(3)the expression of VEGF in cells between high-glucose group and low-glucose group after berberine treatment at different concentrations was significantly different(P<0.05)through pair-to-pair comparison.Conclusion:(1)Compared with low glucose environment,high glucose environment promoted the cell proliferation of retinal vascμLar endothelial cells and increased the expression of VEGF in HRCECs.(2)berberine showed a significant dose-and time-dependent inhibitory in the proliferation of HRCECS cμLtured in a high-glucose environment.(3)berberine down-regμLated the expression of VEGF in HRCECS grown in a high glucose environment,and it was positively correlated with the concentration and time.