摘要
目的探讨基质金属蛋白酶9（MMP-9）及其组织抑制剂-1（TIMP-1）在脂多糖（LPS）诱导内皮细胞损伤中的表达，并观察肝素对其水平的影响。方法LPS10txg／ml刺激人肺微血管内皮细胞（HPMEC）诱导损伤，肝素预处理组分别于LPS刺激前15min加入0．1U／ml及1U／ml普通肝素，对照组加入等量磷酸盐缓冲液（PBS）。分别在刺激2、6、12h收集细胞提取RNA，应用实时荧光定量逆转录-聚合酶链反应（qRT—PCR）检测各组细胞中MMP～9及TIMP-1的mRNA表达。结果与对照组比较，LPS刺激2h组MMP-9和TIMP-1的mRNA表达明显增高，12h达高峰（MMP-9mRNA：4．26±0．81比1．00±0．46，TIMP-1mRNA：4．93±0．08比1．00±0．13，均P〈0．05），以TIMP-1增高明显。预防性应用肝素可明显降低MMP-9和TIMP-1的mRNA表达（0．1U／ml组：MMP-9mRNA2．74±0．30，TIMP-1mRNA2．96±0．13；1U／ml组iMMP-9mRNA3．08±0．48，TIMP-1mRNA2．93±0．27，均P〈0．05）。不同剂量肝素组间基因表达水平差异无统计学意义。结论LPS诱导内皮细胞损伤时MMP-9、TIMP-1表达增加，肝素可能通过调节MMP-9、TIMP-1表达水平发挥其保护作用。
Objective To investigate the expressions of matrix metalloproteinase-9 and tissue inhibitors of metalloproteinase-1 （TIMP-1 ） in injured endothelial cells induced by lipopolysaccharide （LPS） and the effects of unfraetionated heparin （UFH） on the level of expressions. Methods The human pulmonary microvascular endothelial cells （HPMECs） were injured by LPS （10μg/ml）. In UFH pretreatment group, the cells were interfered with 0.1 U/ml or 1 U/ml UFH within 15 minutes before stimulus of LPS. In control group, the cells were cultured in equal volume of phosphate buffered saline （PBS）. The RNA of the respectively cells were extracted at 2, 6, 12 hours after stimulus, and the expressions of MMP-9 mRNA and TIMP-1 mRNA were detected by real-time quantitative reverse transcription-polymerase chain reaction （qRT-PCR）. Results Compared with control group, the expressions of MMP-9 mRNA and TIMP-1 mRNA were increased after stimulation of LPS, and peaked at 12 hours （MMP-9 mRNA： 4.26 ± 0.81 vs. 1.00 ± 0.46, TIMP-1 mRNA： 4.93 ± 0.08 vs. 1.00 ± 0.13, both P〈0.05）, the change in TIMP-1 was more significant. While as UFH pretreatment could significantly down-regulated the mRNA expressions of MMP-9 and TIMP-1 （UFH 0.1 U/ml group MMP-9 mRNA ： 2.74 ± 0.30, TIMP-1 mRNA ： 2.96 ± 0.13 ; UFH 1 U/ml group MMP-9 mRNA： 3.08 -± 0.48, TIMP-1 mRNA： 2.93 ± 0.27, all P〈O.05）. There were no significant differences in mRNA expressions between two UFH groups. Conclusions The expressions of MMP-9 and TIMP-1 of HPMEC injured by LPS were obviously increased, UFH might attenuate the injury via inhibiting the expressions of MMP-9 and TIMP-1.
出处
《中国危重病急救医学》
CAS
CSCD
北大核心
2012年第8期490-492,共3页
Chinese Critical Care Medicine
基金
辽宁省科技厅药物源头创新研究课题（20071027-7）
辽宁省医学高峰建设工程项目（2010-1067）
辽宁省沈阳市科技计划项目（1710-205-1-01）
关键词
脂多糖
内皮细胞
普通肝素
基质金属蛋白酶9
金属蛋白酶组织抑制剂-1
Lipopolysaccharide
Endothelial cell
Unfractionated heparin
Matrix metalloproteinase-9
Tissue inhibitors of metalloproteinase-1
