ABSTRACT
Objective:To observe the effects of salvianolic acid B on migration and tube formation of the retinal vascular endothelial cell (RVEC) in high glucose, and explore its mechanism with network pharmacology.Methods:The cells were divided into normal group, model group and 1.0, 0.5, 0.1 μg/ml salvianolic acid B group according to the random number table method. The cells of each group were added with 5.5 mmol/L glucose for intervention, and the salvianolic acid B group was added with 1.0, 0.5, and 0.1 μg/ml salvianolic acid B for intervention. After 72 h, the cell viability of each group was detected by the CCK-8 method. The cells were divided into normal group, model group and low-, medium-, and high-dose salvianolic acid B group according to the random number table method. Then the cells of the normal group were added with 5.5 mmol/L glucose; the model group was added with 25 mmol/L glucose; the low-, medium-, and high-dose salvianolic acid B group was added with 25 mmol/L glucose and 0.062 5, 0.1250, 0.250 0 μg/ml salvianolic acid B. Then by taking Transwell test to detect the number of cell migration, and Matrigel test to analyze the total length of cells tubes. The active targets of Salvianolic acid B were screened by SuperTarget and Swiss TargetPrediction. Then, the targets of diabetic retinopathy were obtained by searching the GAD database, pharmGkb database, TTD database, DiGSeE database and OMIM database. The effective targets of drug-disease interaction were screened, and the component-target-disease interaction network was constructed by Cytoscape. Finally, the effective targets were analyzed by DAVID for GO analysis and KEGG pathway enrichment analysis. Molecular docking was performed by using Accelrys Discovery Studio Client 2.5 software.Results:The CCK-8 method showed that the cell absorbance values of 0.5 and 0.1 μg/ml salvianolic acid B group were not significantly different from those of the normal group ( P>0.05). The results of Transwell experiment and Matrigel experiment showed that compared with the model group, the relative number of migrating cells and the total length of tubule formation in each dose group of salvianolic acid B decreased ( P<0.05 or P<0.01). The interaction network revealed that salvianolic acid B acted on 46 targets and 8 signaling pathways. Conclusions:Salvianolic acid B could inhibit the migrating and tube forming ability of RVEC cultivated by high glucose. The results suggest that salvianolic acid B may play roles in preventing diabetic retinopathy.
ABSTRACT
Objective To isolate and identify 3 flavonoids (taxifolin, orobol and quercetin) from Cudrania tricuspidata, and develop a method for determining 3 flavonoid constituents in Cudrania tricuspidata. Methods Three flavonoids was isolated from ethanol extract of Cudrania tricuspidata by chromatography, and its structure was identified by nuclear magnetic resonance. The analysis was conducted on an Aglient C18 column (4.6 mm ×250 mm, 5 μm) eluted with 1％ acetic acid and methanol as mobile phases in gradient mode. The flow rate was 1 ml/min and the detection wavelength was set at 310 nm. The column temperature was 25 ℃. Results Taxifolin, orobol and quercetin were isolated from ethanol extract of Cudrania tricuspidata by chromatography. The content of taxifolin, orobol and quercetin were 0.850 mg/g, 0.518 mg/g, 0.103 mg/g. Conclusion The method can be used for the quality control of Cudrania tricuspidata as a reference.