Anderson S S, Wu K, Nagase H, Stettler-Stevenson W G, Kim Y, Tsilibary E C
Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis 55455, USA.
Cell Adhes Commun. 1996 Aug;4(2):89-101. doi: 10.3109/15419069609010765.
Human mesangial cells grown in either 5 or 25 mM glucose were cultured on type IV collagen which had been previously control-incubated or in vitro glycated. Northern blot analysis revealed that after 3-7 days in culture mesangial cells on glycated type IV collagen expressed approximately 25-200% more alpha 1 (IV), approximately 20-50% less matrix metalloproteinase 2 (MMP-2), and 65-75% more tissue inhibitor of metalloproteinase 1 (TIMP-1). Decreased immunoreactivity (approximately 30-40%) and collagenolytic activity (approximately 10-40%) corresponding to MMP-2 was also detected in media conditioned during the third day of culture on glycated type IV collagen. These effects on cell function were related to the extent of type IV collagen modification and were similar for cells cultured in 5 or 25mM glucose. Elevated glucose (25 vrs 5 mM) increased expression of alpha 1 (IV) mRNA (approximately 40-70%) and in conjunction with matrix glycation resulted in detectable levels of MMP-9 message by northern blot although collagenolytic activity corresponding to MMP-9 was not detectable by zymography. We conclude that glucose and matrix glycation may each alter mesangial cell function, perhaps leading to an imbalance in mesangial matrix synthesis and degradation which could contribute to mesangial expansion characteristic of diabetic renal disease.
在5 mM或25 mM葡萄糖中培养的人系膜细胞，被培养在预先经对照孵育或体外糖基化的IV型胶原上。Northern印迹分析显示，在培养3 - 7天后，在糖基化IV型胶原上的系膜细胞表达的α1(IV)多约25 - 200%，基质金属蛋白酶2(MMP - 2)少约20 - 50%，金属蛋白酶组织抑制剂1(TIMP - 1)多65 - 75%。在糖基化IV型胶原上培养第三天的条件培养基中，也检测到与MMP - 2相应的免疫反应性降低(约30 - 40%)和胶原olytic活性降低(约10 - 40%)。这些对细胞功能的影响与IV型胶原修饰程度有关，对于在5 mM或25 mM葡萄糖中培养的细胞是相似的。高糖(25 mM对5 mM)增加α1(IV) mRNA的表达(约40 - 70%)，并且与基质糖基化一起通过Northern印迹导致可检测水平的MMP - 9信息，尽管通过酶谱法未检测到与MMP - 9相应的胶原olytic活性。我们得出结论，葡萄糖和基质糖基化可能各自改变系膜细胞功能，可能导致系膜基质合成和降解失衡，这可能导致糖尿病肾病特征性的系膜扩张。
