Nitric oxide stimulates the activity of a 72-kDa neutral matrix metalloproteinase in cultured rat mesangial cells
Nitric oxide stimulates the activity of a 72-kDa neutral matrix metalloproteinase in cultured rat mesangial cells
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DOI：
10.1006/bbrc.1996.0125
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发表时间：
1996-01-26
影响因子：
3.1
通讯作者：
Singhai, PC
中科院分区：
文献类型：
--
作者：
Trachtman, H;Futterweit, S;Singhai, PC
我们最近发现，诱导型一氧化氮合酶（iNOS）活性的刺激减少胶原和纤维连接蛋白在培养的大鼠系膜细胞的积累。因此。我们在体外检测了一氧化氮（NO）是否影响这些细胞的72 kDa中性基质金属蛋白酶的活性。在生物素-抗生物素蛋白ELISA中和通过酶谱法评估酶活性。将肾小球系膜细胞暴露于细胞因子干扰素（IFN）-γ和脂多糖（LPS），使明胶溶解活性增加325 ± 60%（P <0.025）。与20 mM L-精氨酸共孵育导致基质金属蛋白酶水平进一步增加。添加L-NAME，一种iNOS抑制剂。逆转了IFN-γ/LPS诱导的明胶分解活性的升高。与外源性NO供体S-亚硝基-N-十六烷基-D，L-青霉胺（SNAP）孵育，金属蛋白酶活性呈剂量依赖性增加（P <0.01）。NO诱导的金属蛋白酶活性的变化也证明了酶谱。这些数据表明，NO调节活性的72 kDa的中性基质金属蛋白酶，并建议，改变NO的生产可能有助于肾小球硬化和肾小管间质纤维化的发展，在慢性肾脏疾病状态。(C)出版社：Academic Press，Inc.
We recently demonstrated that stimulation of inducible nitric oxide synthase (iNOS) activity reduced the accumulation of collagen and fibronectin in cultured rat mesangial cells. Therefore. we examined whether nitric oxide (NO) influenced the activity of a 72 kDa neutral matrix metalloprotrinase by these cells in vitro. Enzyme activity was assessed in a biotin-avidin ELISA and by zymography. Exposure of mesangial cells to the cytokines, interferon (IFN)-gamma and lipopolysaccharide (LPS), increased gelatinolytic activity by 325 +/- 60% (P < 0.025). Co-incubation with 20 mM L-arginine caused a further increase in matrix metalloproteinase levels. Addition of L-NAME, an inhibitor of iNOS. reversed the IFN-gamma/LPS-induced rise in gelatinolytic activity. Incubation with the exogenous No donor, S-nitroso-N-ncetyl-D,L-penicillamine (SNAP), resulted in a dose dependent increase in metalloproteinase activity (P < 0.01). The NO-induced changes in metalloproteinase activity were also demonstrable by zymography. These data indicate that NO modulates the activity of a 72 kDa neutral matrix metalloproteinase and suggest that altered NO production may contribute to the development of glomerulosclerosis and tubulointerstitial fibrosis in chronic renal disease states. (C) 1996 Academic Press, Inc.