游离脂肪酸对肾小管上皮细胞TIMP-1、PAI-1表达及细胞外基质分泌的影响

目的探讨游离脂肪酸（Free fatty acids，FFAs）对人肾小管上皮细胞（renal tubular epithelial cells，RTECs）细胞外基质降解酶抑制剂[tissue inhibitor of metalloproteinase 1（TIMP-1）、plasminogen activator inhibitor 1（PAI-1）]表达及细胞外基质[纤维连接蛋（fibronectin，FN）、胶原]分泌的影响。方法用去脂牛血清白蛋白（defatty bovine serum albumin，d-BsA）为油酸（Oil acids，OA）载体，用不同浓度OA刺激培养RTECs，以不加d-BSA及只加普通培养液的细胞作对照组，应用反转录多聚酶链反应（RT-PCR）检测RTECs TIMP-1mRNA、PAI-1mRNA的表达水平，酶联免疫吸附法（FLISA法）、羟脯氨酸消化法分别检测培养上清FN、胶原的蛋白水平。结果d-BSA组与空白对照组RTECs的细胞外基质降解酶抑制剂（TIMP-1mRNA、PAI-1mRNA）表达及细胞外基质蛋白（FN、胶原）分泌比较差异无显著性（P＞0．05），而不同浓度OA组与空白对照组比较差异有显著性（P＜0．05）；且随着OA浓度增加，对RTECs TIMP-1mRNA、PAI-1mRNA表达以及对FN、胶原蛋白分泌的促进作用有增强趋势。结论FFAs通过促进细胞外基质（FN、胶原）的生成及抑制其降解，使其沉积在肾间质，进而在肾小管间质纤维化发生发展中起着重要的作用。

The effects of free fatty acids on the gene expression of extracellular matrix degradation enzyme inhibitors and the secretion of extracellular matrix in renal tubular epithelial cells

Objective To study the effects of free fatty acids(FFAs) on gene expression of extracellular matrix degradation enzyme inhibitors(tissue inhibitor of metalloproteinase-1(TIMP-1),and plasminogen activator inhibitor-1(PAI-1)) and the secretion of extracellular matrix(fibronectin(FN),collagen) protein in proximal renal tubular epithelial cells(RTECs).Methods OA were carried by defatty bovine serum albumin(d-BSA).Cells were exposed to normal medium and different concentrations of OA which using d-BSA as vector.The gene expression of TIMP-1,PAI-1 were detected by reverse transcripase polymerase chain reaction(lRT-PCR),the excretion of FN and collagen protein were assessed by ezyme link immunosordent assay(ELISA) and Hydroxyproline digestion method respectively.Results There was no differences in the gene expression of TIMP-1,PAI-1 and the excretion of FN and collagen protein between cells exposed to normal medium and medium just with OA vector.OA can significantly up-regulated the gene expression of TIMP-1 and PAI-1 as well as the secretion of FN and collagen protein(P<0.05).Conclusion FFAs may promote ECM deposition via promoting ECM production and inhibiting ECM catabolism by RTECs.