辛伐他汀对香烟烟雾提取物诱导的人脐静脉内皮细胞表达sEPCR和mEPCR的影响

Effects of simvastatin on cigarette smoke extract-induced sEPCR and EPCR expression in human umbilical vein endothelial cells

目的:研究辛伐他汀对香烟烟雾提取物(CSE)诱导的人脐静脉内皮细胞(HUVECs)表达可溶性内皮细胞蛋白C受体(sEPCR)和膜联内皮细胞蛋白C受体(mEPCR)的干预作用。方法:体外培养的4～6代HU-VECs随机分为对照组、5%CSE组、不同浓度辛伐他汀组及辛伐他汀干预组,辛伐他汀组分别加入50、100、200μmol/L辛伐他汀液孵育24 h,辛伐他汀干预组先以50、100、200μmol/L辛伐他汀预处理细胞2 h,再与5%CSE孵育24 h。收集各组细胞及上清液,ELISA法检测上清液中sEPCR蛋白含量,实时定量PCR法检测各组细胞中mEPCR mRNA的表达。结果:(1)5%CSE组sEPCR蛋白含量高于对照组,mEPCR mRNA表达低于对照组,差异均有统计学意义(均P<0.05);(2)100μmol/L与200μmol/L辛伐他汀组sEPCR蛋白含量均高于对照组,低于5%CSE组,其mEPCR mRNA表达均低于对照组,高于5%CSE组,差异均有统计学意义(均P<0.05);(3)各辛伐他汀干预组sEPCR蛋白含量均低于5%CSE组,但高于对照组及相应浓度的辛伐他汀组;相反,各辛伐他汀干预组mEPCR mRNA表达均高于5%CSE组,低于对照组及相应浓度的辛伐他汀组,差异均有统计学意义(均P<0.05)。结论:在体外,辛伐他汀通过上调HUVECs mEPCR mRNA的表达,降低sEPCR的分泌,对CSE介导的内皮细胞凝血功能障碍可能具有一定的改善作用。

AIM： To investigate the effects of simvastatin on cigarette smoke extract（CSE）-induced expression levels of soluble endothelial cell protein C receptor（sEPCR） and membrane-associated endothelial cell protein C receptor（mEPCR） in human umbilical vein endothelial cells（HUVECs）.METHODS： Cultured HUVECs at passage 4 to 6 were randomly divided into control group,5% CSE group,simvastatin groups and simvastatin＋CSE groups.In simvastatin groups,HUVECs were incubated with simvastatin at the concentrations of 50,100 and 200 μmol/L for 24 h.In simvastatin＋CSE groups,the cells were treated with simvastatin at the concentrations of 50,100 and 200 μmol/L for 2 h,and then exposed to CSE for 24 h.The protein level of sEPCR in the culture supernatants was measured by ELISA.The cells were collected for determining the mRNA expression of mEPCR by real-time PCR.RESULTS： Compared with control group,the protein level of sEPCR was significantly increased,and the mRNA expression of mEPCR was significantly decreased in 5% CSE group（both P0.05）.The protein levels of sEPCR were significantly increased,and the mRNA expression of mEPCR was significantly decreased in 100 μmol/L and 200 μmol/L simvastatin groups.However,the protein levels of sEPCR were lower,and the mRNA expression of mEPCR was significantly higher in 100 μmol/L and 200 μmol/L simvastatin groups than those in 5% CSE group.Compared with 5% CSE group,the protein levels of sEPCR in simvastatin＋CSE groups were significantly decreased,but higher than those in control group and simvastatin group with corresponding concentration.On the contrary,the mRNA expression of mEPCR in simvastatin＋CSE groups was significantly increased,but lower than that in control group and simvastatin group with corresponding concentration（all P0.05）.CONCLUSION： Simvastatin obviously increases the mRNA expression of mEPCR,decreases the protein level of sEPCR,and attenuates the CSE-induced endothelial injury in vitro.