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目的探讨尿毒症毒素对人肾小管上皮细胞外基质的影响及临床意义。方法无菌收集40份尿毒症患者血清和20例正常人血清,培养及鉴定人肾小管上皮细胞株(HK-2),采用消化法检测羟脯氨酸含量,ELISA方法检测纤连蛋白(FN)、金属蛋白酶-1(MMP-1)、金属蛋白酶组织抑制因子-1(TIMP-1)的蛋白分泌量,RT-PCR方法检测MMP-1、TIMP-1 mRNA的表达。结果5%～20%浓度尿毒血清组胶原蛋白、FN分泌量均较正常对照组增高(P＜0．01),且随着尿毒血清浓度的升高而增高,以分子量＞10000ku的组分增高最为显著(P＜0．01)。各浓度尿毒血清组MMP-1/TIMP-1基因表达和蛋白分泌的比值均较正常对照组降低(P＜0．01),且随着尿毒血清浓度的增高而降低,以分子量＞10000ku尿毒血清组降低最为显著,而分子量＜5000ku尿毒血清组较正常对照组增高(P＜0．01)。结论慢性肾衰竭患者体内蓄积的尿毒症毒素可能通过促进人肾小管上皮细胞外基质的分泌,减少其降解,加速肾小管-间质纤维化的进展,其中以分子量＞10000ku的尿毒症毒素作用最为显著。
Objective To investigate the effect of uremic toxins on extracellular matrix of human renal tubular epithelial cells and its clinical significance. Methods Serum samples of 40 patients with uremia and 20 normal controls were collected aseptically. Human renal tubular epithelial cell line (HK-2) was cultured and identified. The content of hydroxyproline was detected by digestion and the content of fibronectin (FN) 1, TIMP-1, MMP-1 and TIMP-1 were detected by RT-PCR and RT-PCR. Results The secretion of collagen and FN in 5% ~ 20% uremic serum group was higher than that in normal control group (P <0.01), and increased with the increase of uremic serum concentration. The fraction of molecular weight> 10000 ku increased The most significant (P <0.01). The ratio of MMP-1 / TIMP-1 gene expression and protein secretion in each concentration of uremic serum was lower than that of the normal control group (P <0.01), and decreased with the increase of serum urease concentration. Group, the most significant reduction, while the molecular weight <5000ku uremia serum group increased compared with the normal control group (P <0.01). Conclusion Uremic toxins accumulated in patients with chronic renal failure may promote the development of tubulointerstitial fibrosis by promoting the secretion of extracellular matrix of human renal tubular epithelial cells, reducing their degradation and accelerating the progress of tubulointerstitial fibrosis. The effect of uremic toxins with molecular weight> 10000ku is the most Significant.