摘要
目的研究氯化高铁血红素(Hemin)对非对称二甲基精氨酸(ADMA)诱导的血管内皮损伤的保护作用及其与诱导型一氧化氮合酶(iNOS)和基质金属蛋白酶-9(MMP-9)的关系。方法体外培养人脐静脉内皮细胞(HUVECs),MTT法检测Hemin(5～20μmol/L)对ADMA(3～30μmol/L)诱导细胞损伤的影响,Western blot法检测胞内MMP-9蛋白表达水平,RT-PCR技术评价MMP-9、iNOS及单核细胞趋化因子-1(MCP-1)mRNA的表达量,ELISA法检测细胞上清液肿瘤坏死因子α(TNF-α)水平变化。结果 ADMA(3～30μmol/L)能使细胞活性降低,其损伤呈剂量依赖性,10μmol/L的ADMA显著升高了MMP-9、iNOS、MCP-1和TNF-α的表达水平(P<0.01)。而10μmol/L的Hemin预处理能明显干预ADMA对细胞活性的影响,并能下调ADMA诱导的MMP-9、iNOS、MCP-1和TNF-α的表达,与对照组相比差异具有显著性(P<0.01)。结论 Hemin对ADMA所致内皮细胞损伤有保护作用,且与MMP-9和iNOS水平降低有密切关系。
【Objective】 The present study aimed to test whether beneficial effect of Hemin on the endothelium induced by asymmetric dimethylarginine（ADMA） is related to the process that changing the level of matrix metalloproteinase 9（MMP-9） and nitric oxide synthase（iNOS）,and further to explore the molecular mechanism.【Methods】 Cultured human umbilical vein endothelial cells（HUVECs） in vitro.The cell viability was tested by MTT to detect the effects of Hemin（5～20 μmol/L） on cells induced by ADMA（3～30 μmol/L）.The protein expression of MMP-9 was detected by western blot.The mRNA expression of MMP-9,iNOS and monocyte chemoattractant protein 1（MCP-1） was tested by RT-PCR.The level of TNF-α in the conditioned medium was analyzed by ELISA.【Results】 The activation of endothelial cells which induced by different concentration of ADMA for 24 h decreased greatly.In the same time,the impact of ADMA on the cell viability was dose dependent.Incubation with ADMA（10 μmol/L） for 24 h elevated the mRNA level of MMP-9,iNOS and MCP-1,and the protein level of MMP-9 and TNF-α.These effects could be attenuated by pretreatment with Hemin（10 μmol/L）.【Conclusions】 The Protective effects of Hemin on ADMA-induced endothelial injury maybe related to the process that inhibiting the expression of MMP-9 and iNOS.
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2011年第15期1791-1796,共6页
China Journal of Modern Medicine
基金
国家自然科学基金资助项目(No:30871200)
江苏省高等学校大学生实践创新训练计划资助项目(No:KY208J2010002)
