高糖培养的大鼠肾小球系膜细胞中NO的变化及其对细胞外基质的影响

Changes in nitric oxide production in high glucose cultured rat mesangial cells and its effects on extracellular matrix

目的 探讨高糖培养的肾小球系膜细胞中一氧化氮（NO）合成的变化,以及上述变化对细胞外基质的影响.方法 体外培养大鼠肾小球系膜细胞,按干预方法分为正常组（NG组,5.56 mmol·L-1 DMEM）,高糖组（HG组,25 mmol·L-1 DMEM）、一氧化氮供体组（SNP组,5.56 mmol·L-1 DMEM＋100 μmol·L-1 SNP）、NO清除剂组（C-PTIO 组,25 mmol·L-1 DMEM＋200 μmol·L-1 C-PTIO）,以Griess比色法测定培养上清中NO水平;用ELISA法测定培养上清中Ⅳ型胶原、层黏蛋白含量.结果 高糖培养条件下上清中NO增多（P〈0.05）,Ⅳ型胶原、层黏蛋白含量增加（P〈0.01）,应用C-PTIO可显著降低上清中NO的含量,Ⅳ型胶原、层黏蛋白的含量也降低.结论 高糖可诱导大鼠肾小球系膜细胞产生NO,异常增多的NO又可导致系膜细胞中Ⅳ型胶原、层黏蛋白的积聚.

Objective To explore changes in nitric oxide （NO） production in high glucose cultured rat mesangial cells （MCs） and its effect on extraeellular matrix. Methods The rat MCs were cultured in vitro and then divided into a normal group （ NG group, 5.56 mmol/L DMEM） , a high glucose group （ HG group, 25 mmol/L DMEM）, a NO donator group （SNP group, 5.56 retool/1 DMEM ＋ 100 μ mol/L SNP） and a MO clearance group （C -PTIO group, 25 mmol/L DMEM ＋ 200 μmol/L C -PTIO ）. The NO level in the supernatant was determined by the Griess method. The levels of laminin and collagen 1V was determined by the ELISA method. Results Under high glucose condition, the amounts of NO, collagen IV and laminin in the supernatant were increased （P 〈 0.05 or P 〈0. 01 ） , which were then reduced by the addition of C - PTIO. Conclusion High glucose can induce rat MCs to produce more amounts of NO. Excessive NO can result in the accumulation of collagen IV and laminin in the cells.