Effect Of Calycosin On The Synthesis Of PGI2 And TXA2 In Vascular Endothelial Cells Induced By Angâ…¡

Objective:Chronic kidney disease(CKD) has been paid more and more attentions because of its high incidence,high medical costs and poor prognosis. Modern medical studies have shown that endothelial dysfunction plays an important role in the occurrence and development of CKD.Moreover,endothelial dysfunction is also involved in the pathophysiology of cardiovascular disease. Therefore, the positive and effective protection of endothelial function not only slows the progress of CKD, but also reduces the incidence of cardiovascular complications of CKD patients. The research about prevention of endothelial injury by Chinese medicine has become an important research area in recent years, and studies have reported that astragalus has a protection of endothelial cells. Prostacyclin (PGI2) and thromboxane A2 (TXA2) secreted by the endothelial cells are a pair of prostaglandin with opposite effect, which are related with thromboembolic disease and the occurrence of cardiovascular events. AngⅡis a major effector molecule in RAS system, which can cause endothelial dysfunction. Therefore,cultured human umbilical vein endothelial cells (HUVECs) were induced by angiotensinⅡ(AngⅡ).To investigate the effect of calycosin on the synthesis of prostacyclin (PGI2) and thromboxane A 2(TXA 2) in human umbilical vein endothelial cells (HUVECs) induced by AngⅡ,and explicit the protection of calycosin on the endothelial,in order to supply the evidence for the further development of Astragalus.Methods:Human umbilical endothelium derived cells were used in this study, which were cultured with L-DMEM medium in 5%CO2,37℃to sufficient number. HUVECs were digested with 0.25% trypsin after washing with PBS and terminated by serum-free L-DMEM medium. Then HUVECs were seeded in 24-well culture plate (500ul/well) by 2×105/mL and incubated in 5% CO2,37℃, finally supernatants were removed after 24 hours. HUVECs were grouped as follows①control;②AngⅡ(1×10-6 moL·L-1);③AngⅡ(1×10-6 moL·L-1)+ calycosin (0.1 mg·L-1);④AngⅡ(1×10-6 moL·L-1)+calycosin (1mg·L-1);⑤AngⅡ(1×10-6 moL·L-1)+calycosin (10mg·L-1).Supernatants were collected after cultured 0,6,12,18,24h. The levels of PGI2 and TXA2 were detected by ELISA respectively. Results:①Compared with controls, AngⅡ(1×10-6 moL·L-1) promoted the synthesis of PGI2 and TXA2,futhermore the synthesis were in time-dependent manner. (r=0.891 P=0.043) (r=0.95 P=0.013) (P<0.05);②Compared with AngⅡgroup, calycosin (0.1 mg·L-1)inhibited the production of PGI2 and TXA2 in HUVECs induced by AngⅡ; calycosin (1mg·L-1) promoted the production of PGI2,while inhibited the production of TXA2 in HUVECs induced by AngⅡ; calycosin (10mg·L-1) promoted the production of PGI2 and TXA2 in HUVECs induced by AngⅡ.Conclusion:HUVECs can synthesize PGI2 and TXA2 induced by AngⅡ. Calycosin extracted from Astragalus may inhibit the synthesis of PGI2 and TXA2 in HUVECs induced by AngⅡ. Possibly effects are mediated partly through reducing the activity of NF-κB. It is proposed that calycosin may affect the production of PGI2 and TXA2 in normal endothelial cells.