Protective Effect of Huoxue Huayu Prescription on Retinal Capillary Pericytes of Diabetic Rats

OBJECTIVE To determine the protective effect of Huoxue Huayu prescription on retinal capillary pericytes(RCPs) of diabetic rats. METHODS Forty SD rats were conducted and randomly separated into 4 groups, ten of them were in the normal group, while other 30 rats were divided into Huoxue Huayu prescription high-dose diabetes group(6.75 g·kg^-1, n=10), low-dose diabetes group(3.375 g·kg^-1, n=10) and model group(n=10) after successfully established diabetic model by intraperitoneal injection of streptozotocin. Retina was taken after gastric administration for 60 d for RCPs immunohistochemical study, while Western blot assay was adopted to test the Bax, Bak, Bcl-2, caspase-3 protein expression. In vitro assay, RCPs were incubated with high glucose(30 mmol·L^-1) and intervened by different concentrations of Huoxue Huayu prescription. CCK-8 assay was employed to measure the relative growth rate. Apoptosis of RCPs was observed with a fluorescence microscope using JC-1 stain, and quantitied by flow cytometry using Annexin V-FITC stain. RESULTS More obvious RCPs apoptosis were observed in diabetic model rats than that in normal group, and the Huoxue Huayu prescription could inhibit the apoptosis of RCPs. Compared with 0 μg·mL^-1, the proliferation in 25-100 μg·mL^-1 of Huoxue Huayu prescription increased significantly(P<0.05 or P<0.01). There was a significant difference between 25-100 μg·mL^-1 and 0 μg·mL^-1 in anti-apoptotic effect induced by high glucose. CONCLUSION Huoxueguayu prescription can inhibited high glucose induced apoptosis in RCPs, and the preliminary mechanism may be associated with major outer membrane proteins/BCL-2 pathway.