Neferine Protects Endothelial Cells Against Damages Induced By LPC And Its Mechanisms

Objective: To investigate the protective effect of neferine againstdamages of endothelial cells induced by lysophosphatidylcholine(LPC)and to study the underlying mechanisms.Methods: The human umbilical vein endothelial cells(HUVECs)were treated with LPC (10μg/ml) for 24h to establish the model ofendothelial cells injury; HUVECs were prior exposed to neferine(0.1, 1.0or 10.0μmol/l) for 1h, and then exposed to LPC in the presence of thedrugs for 24h in all; And also we set the control group and theneferine(10.0μmol/l)group; At the end of the experiment, the culturedmedium was collected for measuring the concentration of lactatedehydrogenase(LDH), nitric oxide(NO), maleic dialdehyde(MDA) aswell as asymmetric dimethylarginine(ADMA), and the cells werecollected for measuring the intracellular reactive oxygen species(ROS)level. The transudation content of LDH was determined by colorimetryreaction; The content of NO was measured by nitrate reductase method;The level of MDA was measured by thiobituric acid reaction(TBA)method; The concentration of ADMA was measured by high performanceliquid chromatography(HPLC); The intracellular ROS level was detectedby fluorospectrophotometer analysis.Results: 1.Compared with control group, exposure of endothelial cells to LPC (10μg/ml) for 24h significantly increased the concentrationof LDH(P＜0.05), MDA(P＜0.05), ADMA(P＜0.05) in the medium and theintracellular ROS level(P＜0.05), concomitantly significantly decreasedthe concentration of NO(P＜0.05) in the medium. 2.Compared with LPCgroup, neferine (0.1, 1.0 or 10.0μmol/l) significantly inhibited theelevated concentration of LDH(P＜0.05), MDA(P＜0.05), ADMA(P＜0.05)as well as the intracellular ROS level(P＜0.05), concomitantlysignificantly attenuated the decreased level of NO(P＜0.05) induced byLPC. 3. Neferine (10.0μmol/l) itself had no effect on the concentration ofLDH(P＞0.05), NO (P＞0.05), MDA(P＞0.05) as well as ADMA (P＞0.05)in the medium and the intracellular ROS level(P＞0.05).Conclusions: 1.Neferine protected endothelial cells against damagesinduced by LPC. 2.The protective effect of neferine on the endothelialcells is related to antagonize lipid peroxidation and to decrease the levelof ADMA.