中文摘要：

目的：探讨晚期糖基化终产物通过MAPK信号转导通路引起人脐静脉内皮细胞骨架结构改变的作用机制。方法：用胰酶消化法分离培养人脐静脉内皮细胞，以不同浓度的晚期糖基化终产物修饰的人血清白蛋白（AGE-HSA）与之作用不同时间，并选用不同的试剂以及主导激活或抑制的腺病毒突变体以阻断或激活MAPK通路，与对照组进行比较，采用免疫荧光染色法显示细胞骨架肌动蛋白F-actin的结构改变。结果：AGE-HSA以剂量和时间依赖的方式引起内皮细胞骨架蛋白F-actin结构的改变，未经修饰的HSA无此作用；p38通路的特异性抑制剂SB203580和ERK通路的特异性抑制剂PD98059均可抑制AGEs对细胞骨架的作用，而JNK通路抑制剂SP600125则无此作用；MKK6b、MEK1结构活性型突变体的重组腺病毒可明显诱导细胞骨架改变，MKK7活性型则无此作用；且MKK6b、MEK1无活性型突变体可明显抑制AGEs对细胞骨架的作用；p38α亚型的无活性突变体可阻断MKK6b结构活性型突变体诱导的细胞骨架改变。结论：AGE修饰蛋白通过MAPK家族中的p38和ERK信号转导通路引起内皮细胞骨架肌动蛋白F-actin的重组和再分布。

英文摘要：

AIM： To investigate the effect of advanced glycosylation end products （AGEs） modified protein on morphological changes of aetin eytoskeleton in primary endothelial cells and the role of MAPK signaling pathways in this pathological procedure. METHODS： Human umbilical vein endothelial cells （HUVECs） were incubated with AGEs modified human serum albumin （ AGE - HSA） at concentrations of 12. 5,25, 50, and 100 mg/L, respectively, for 2, 4, 8, 12 and 24 hours. The cells were treated with different chemical compounds of inhibitors, or adenoviral deliver approach （ either dominant positive or negative adenoviral constructs） of MAP kinases to specifically block or activate certain signal transduction pathways under above situations. As control, HSA of the same concentration was administered to cells at the same time. The treated cells were incubated with FITC - phalloidin to stain F - actin. RESULTS： F - actin in HUVECs was rearranged greatly by AGEs in a concentration and time - dependent manners. The unmodified HSA did not influence F - actin distributions. The AGEs - induced changes were blocked by pretreatment with SB203580, PD98059 for 30 rain, or pre - infection with recombinant virus of dominant negative form of MKK 6b [ MKK6b （ A ） ], MEK1 [ MEK1 （ A ） ] for 24 h, while SP600125 and dominant negative form of MKK7 [ MKK7 （A） ] failed to inhibit the effects of AGEs. Furthermore, the infecion of recombinant virus of constitutive active form of MKK6b [ MKK6b （E） ] or MEKI [ MEK1 （E） 1 could also induced rearrangement of F- actin, respectively, while the effect elicited by [ MKK6b （E）] was abolished by co-infection with recombinant adeno -virus of dominant negative p38α. CONCLUSION： AGEs -induced morphological changes of F -actin in endothelial cells are mediated by p38 -and ERK MAPK signal pathways.