Elliot S J, Striker L J, Stetler-Stevenson W G, Jacot T A, Striker G E
Renal Cell Biology Section, Metabolic Disease Branch, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA.
J Am Soc Nephrol. 1999 Jan;10(1):62-8. doi: 10.1681/ASN.V10162.
Glomerulosclerosis is characterized by extracellular matrix accumulation and is often associated with mesangial cell proliferation. Heparin-like molecules have been shown to decrease glomerulosclerosis in vivo, although their cellular site and mechanism of action is still unclear. In this study, a line of glomerular mesangial cells derived from normal mice was used to determine whether pentosan polysulfate (PPS) inhibited proliferation and altered extracellular matrix turnover. Cells treated with PPS showed a decrease in cell number beginning 24 h after treatment, which was maintained for 5 d. For matrix accumulation and degradation studies, cells were treated for 5 d and collagen types I and IV protein were measured by enzyme-linked immunosorbent assay as well as matrix metalloproteinases (MMP) measured by zymography. Collagen types 1 and type IV were significantly decreased in the media (P < 0.0001) and cell layer (P < 0.005) after treatment with PPS but not after treatment with heparin. By zymography, MMP-2 was significantly increased after treatment with PPS (P < 0.001) and heparin (P < 0.05). PPS and heparin also decreased MMP-9 (P < 0.001) after treatment. Reverse zymography showed the presence of tissue inhibitors of metalloproteinases (TIMP)-1 and -2 in control mesangial cells. Treatment with PPS and heparin increased TIMP-1. In addition, TIMP-3 was found in the medium of treated but not control cells. In conclusion, PPS alters extracellular matrix turnover through the induction of MMP-2 and alterations in the TIMP profile and may be useful in decreasing progressive glomerulosclerosis.
肾小球硬化的特征是细胞外基质积聚，且常伴有系膜细胞增殖。尽管类肝素分子在体内的细胞作用位点和作用机制仍不清楚，但已表明其可减轻肾小球硬化。在本研究中，使用源自正常小鼠的肾小球系膜细胞系来确定戊聚糖多硫酸盐（PPS）是否抑制增殖并改变细胞外基质的周转。用PPS处理的细胞在处理后24小时开始细胞数量减少，并持续5天。对于基质积聚和降解研究，细胞处理5天，通过酶联免疫吸附测定法测量I型和IV型胶原蛋白，以及通过酶谱法测量基质金属蛋白酶（MMP）。用PPS处理后，培养基（P < 0.0001）和细胞层（P < 0.005）中的I型和IV型胶原蛋白显著减少，但用肝素处理后未减少。通过酶谱法，用PPS处理（P < 0.001）和肝素处理（P < 0.05）后，MMP-2显著增加。处理后PPS和肝素也降低了MMP-9（P < 0.001）。反向酶谱法显示对照系膜细胞中存在金属蛋白酶组织抑制剂（TIMP）-1和-2。用PPS和肝素处理增加了TIMP-1。此外，在处理过的细胞培养基中发现了TIMP-3，而对照细胞培养基中未发现。总之，PPS通过诱导MMP-2和改变TIMP谱来改变细胞外基质的周转，可能有助于减轻进行性肾小球硬化。
