Effect of apigenin on amelioration of Clinopodium chinense for vascular endothelial cell injury induced by high glucose / 中草药

ABSTRACT

Objective: In this study, we specifically deleted apigenin (AP) from the active fraction of Clinopodium chinense (CCE), and aimed at identifying the effect of AP on how CCE exerted its amelioration on high glucose-induced injury in human umbilical vein endothelial cells (EA.hy926). Methods: By using Sephadex LH-20 gel column chromatography, AP was specifically deleted from CCE, and the apigenin-depleted sample of CCE (CCEAP-)was obtained. The cultured endothelial cells were divided into five groups normal control group, high glucose model group, CCE group, CCEAP- group, and AP group. The cell viability was assayed by MTT assay. Flow cytometry was used to measure the intracellular reactive oxygen species (ROS). Morphology of cell apoptosis was determined by fluorescence microscopy with Hoechst staining. The rate of apoptosis was measured by flow cytometry staining with AnnexinV-FITC. Caspase-3 activity was measured using caspase-3 colorimetric assay kit. The expression of Bax was detected by Western blotting. Results: CCE and AP could significantly improve the cell viability, reduce the generation of intracellular ROS in EA.hy926 induced by high glucose. Meanwhile, CCE and AP can reduce the ratio of apoptosis, activity of caspase-3, and expression of Bax, while CCE knocked out apigenin (CCEAP-) had a slight improvement on the high glucose-induced endothelial cell injury. Conclusion: AP is one of the principal components improving the high glucose-induced endothelial cell injury, and its anti-apoptosis effect may be related to anti-oxidative stress.