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目的探讨铅诱导人脐静脉内皮细胞损伤效应及机制。方法用不同浓度的铅染毒人脐静脉内皮细胞,观察细胞形态和活性,通过检测细胞上清液丙二醛(MDA)、非对称性二甲基精氨酸(ADMA)、一氧化氮(NO)的含量和一氧化氮合酶(NOS)的活性等指标,观察铅对人脐静脉内皮细胞损伤的量-效关系(终浓度分别为1、2、4、8mg/L的铅处理细胞24h)和时-效关系(终浓度为4mg/L的铅分别于处理0、3、6、12、24、48h后观察检测指标)。实验分为正常对照组(加入与处理组等体积的D-Hank’s液,铅浓度为0mg/L)、铅处理组(加终浓度分别为1、2、4、8mg/L的铅)、铅&VitC组(铅终浓度为4mg/L,VitC终浓度为100mg/L)和阳性对照组(加终浓度为500μg/L的过氧化氢)。结果 2～8mg/L的铅能显著改变细胞形态、降低细胞活性;能显著性诱导内皮细胞MDA升高。2～8mg/L的铅还能显著性诱导内皮细胞NO含量降低,具有显著的时间依赖性,各处理组间两两比较差异有统计学意义(P<0.05或P<0.01)。4mg/L铅处理时细胞ADMA含量最高,且分别在处理后24h到达最高峰。结论 2～8mg/L铅能显著诱导人脐静脉内皮细胞损伤;ADMA在铅诱导人脐静脉内皮细胞损伤中有着重要作用。
Objective To investigate the effect and mechanism of lead-induced injury of human umbilical vein endothelial cells. Methods Human umbilical vein endothelial cells (HUVECs) were exposed to different concentrations of lead to observe the morphology and activity of the cells. The levels of malondialdehyde (MDA), asymmetric dimethylarginine (ADMA), nitric oxide NO) content and nitric oxide synthase (NOS) activity and other indicators to observe the dose-effect of lead on human umbilical vein endothelial cell injury (final concentrations were 1, 2, 4, 8mg / L of lead-treated cells 24h) and the time-effect relationship (the final concentration of 4mg / L of lead were treated at 0,3,6,12,24,48h after the observation indicators). The experiment was divided into normal control group (equal volume of D-Hank’s solution added and treated group, lead concentration 0mg / L), lead treated group (added lead concentration 1,2,4,8mg / L), lead & VitC group (final lead concentration 4 mg / L, final VitC concentration 100 mg / L) and positive control group (hydrogen peroxide at a final concentration of 500 μg / L). Results Lead (2 ~ 8 mg / L) could significantly change cell morphology and cell viability, and could significantly induce the elevation of MDA in endothelial cells. Lead of 2 ~ 8mg / L could significantly reduce the NO level in endothelial cells, which was significantly dependent on time (P <0.05 or P <0.01). When treated with 4 mg / L Pb, the content of ADMA was the highest, reaching the peak at 24h after treatment. Conclusion 2 ~ 8 mg / L lead can significantly induce human umbilical vein endothelial cell injury. ADMA plays an important role in lead-induced injury of human umbilical vein endothelial cells.