糖尿病肾病患者蛋白氧化损伤与血清基质金属蛋白酶-2的相关性分析

目的检测糖尿病肾病患者血清蛋白氧化损伤指标的变化，探讨蛋白氧化损伤在糖尿病肾病发病中的可能机制。方法应用改良后的Witko—Sarsat法和2，4-二硝基苯肼法检测血清晚期氧化蛋白产物（AOPP）水平、蛋白质羰基（PCO）含量，应用ELISA及酶谱法分别检测血清基质金属蛋白酶-2（MMP-2）的表达水平和活性。结果DNl组、DN2组AOPP水平明显高于DM组（78．23±19．30VS61．25±12．13、101．59±30．22VS61．25±12．13，F=41．988，P〈0．01），DNl组、DN2组PCO含量亦明显高于DM组（0．84±0．03VS0．66±0．02、1．05±0．05VS0．66±0．02，F=205．763，P〈0．01），AOPP、PCO与血清MMP-2的表达水平（r=0．460，0．480，P〈0．05）和活性（r=0．385，0．560，P〈0．05）均呈正相关，多元逐步回归分析表明AOPP、PCO为血清MMP-2表达水平（P〈0．05，P〈0．01）和活性（P〈0．01，P〈0．05）的主要影响因素。结论推测蛋白氧化损伤通过改变MMP-2的表达和活性，影响肾脏基质代谢。

Relationship between oxidative protein damage and serum matrix metalloproteinase-2 in diabetic nephropathy

Objective Serum indicators of oxidative protein damage (OPD) were analyzed to explore the effect on renal MMP/TIMP system of OPD in diabetic nephropathy. Methods AOPP levels and PCO levels were measured by modified Witko-Sarsat's method and 2, 4-dinitrophenylhydrazine spectrophotometric method, expression of serum MMP-2 were determined by ELISA, and MMP-2 activity were detected by zymography . Results AOPP levels of group DN1, group DN2 were higher than those of group DM(78.23±19.30 vs 61.25±12.13,101.59±30.22 vs 61.25±12.13,F＝41.988,P＜0.01). PCO levels of group DN1 and group DN2 were higher than those of group DM (0.84±0.03 vs 0.66±0.02,1.05±0.05 vs 0.66±0.02,F＝205.763,P＜0.01). Pearson correlation analysis indicated AOPP and PCO were positively correlated with the expression (r＝0.460,0.480,P＜0.05) and activity (r＝0.385,0.560,P＜0.05) of serum MMP-2. Multiple stepwise regression analysis showed that AOPP and PCO were major influential factors of serum MMP-2 expression (P＜0.05,P＜0.01) and activity(P＜0.01,P＜0.05). Conclusions OPD might be involved in the imbalance of renal matrix metabolism, which was correlated with the development of diabetic nephropathy.