Effect Of Bushen Huoxue Recipe On PEDF And Related Factors In Retinal Müller Cells Under AGEs/anoxic Conditions

Objective: To observe the effect of Bushen Huoxue Recipe on PEDF and related factors in retinal Müller cells under AGEs/anoxic conditions,and to explore its mechanism of action,to provide a certain effect for the clinical application of Bushen Huoxue method in diabetic retinopathy(DR).Methods:(1)SD rat retinal Müller cells were cultured by modified enzyme digestion method;cell identification was performed by HE staining,GFAP and GS immunocytochemical staining.(2)The serum containing Bushen Huoxue Recipe was obtained by the serum pharmacological method of traditional Chinese medicine.(3)The experiment was divided into eight groups: normal control group,normal Chinese medicine intervention group,low AGEs group,high AGEs group,low AGEs Chinese medicine intervention group,high AGEs Chinese medicine intervention group,simulated hypoxia group,and simulated hypoxia Chinese medicine intervention group.(4)After the experimental intervention for 24 h,48h,and 72 h,the cell supernatant was subjected to vascular endothelial growth factor(VEGF),pigment epithelium-derived factor(PEDF)and interleukin-1β by enzyme-linked immunosorbent assay(ELISA).Determination of(IL-1β)content.Results:(1)The concentration of VEGF in the simulated hypoxia group,low AGEs group and high AGEs group was higher than that in the normal control group(P<0.05).At the same time,the VEGF secretion in the hypoxic Chinese medicine intervention group was lower than that in the simulated hypoxia group.P <0.05);At the same time,the AGE concentration of the lower AGEs group was lower in the lower AGEs group(P <0.05).At the same time,the AGE concentration of the AGEs group was lower in the high AGEs group(P <0.05).The VEGF concentration was higher than the previous time(P <0.05).(2)The concentration of PEDF in the simulated hypoxia group,low AGEs group and high AGEs group was lower than that in the normal control group(P <0.05).At the same time,the concentration of PEDF in the hypoxic Chinese medicine intervention group was higher than that in the simulated hypoxia group(P<0.05).<0.05);The concentration of PEDF in the lower AGEs group was higher in the lower AGEs group(P <0.05).At the same time,the higher the AGEs group,the higher the AGEs group,the higher the PEDF concentration(P <0.05);The PEDF concentrations were all lower than the previous time(P < 0.05).(3)The IL-1β concentration in the simulated hypoxia group,low AGEs group and high AGEs group was higher than that in the normal control group(P <0.05).At the same time,the hypoxic Chinese medicine intervention group was compared with the simulated hypoxia group IL-1β.The concentration of IL-1β was decreased in the lower AGEs group(P <0.05).At the same time,the high AGEs group had higher IL-1β concentration in the AGEs group.P <0.05);IL-1β concentration in each group was higher than the previous time(P <0.05).(4)There was a positive correlation between VEGF and PEDF in normal control group(P<0.05).There was no significant correlation between VEGF and PEDF in hypoxia group and low AGEs group(P>0.05).VEGF and PEDF were negatively correlated in high AGEs group.(5)Compared with the normal control group,the ratio of VEGF/PEDF in the hypoxia group,the low AGEs group and the high AGEs group was higher than that in the normal control group.At the same time,the hypoxia group was compared with the hypoxia group.The ratio of PEDF decreased.The ratio of VEGF/PEDF in the high and low AGEs groups was higher.Conclusion:(1)In this experiment,AGEs(final concentrations of 50 mg/L and 100 mg/L)and hypoxia all resulted in increased secretion of VEGF,decreased PEDF,and increased IL-1β in retinal Müller cells,and this change was higher in the high AGEs group.The lower AGEs group is more pronounced.(2)Under the condition of AGEs(final concentration 50 mg/L and 100 mg/L)and hypoxia,the serum containing Bushen Huoxue Recipe can reduce the expression of VEGF and the secretion of IL-1β in retinal Müller cells,and increase the expression of PEDF protein.This may be one of the drug interventions for the prevention and treatment of DR by Bushen Huoxue.(3)In this experiment,VEGF and PEDF in the retinal Müller cells were dynamically balanced in normal state;AGEs(50 mg/L and 100 mg/L final concentration)and hypoxic conditions not only made the dynamic balance of VEGF and PEDF in retinal Müller cells It was broken,and the degree of damage of AGEs to the dynamic balance of VEGF and PEDF in retinal Müller cells was related to the concentration of AGEs.(4)Bushen Huoxue TCM medicated serum can alleviate the imbalance of VEGF and PEDF in AGEs(final concentration 50 mg/L and 100 mg/L)and hypoxia,which may be an important mechanism for the prevention and treatment of DR.