PKC在TNF-α诱导内皮细胞β-1,4-半乳糖基转移酶-Ⅰ表达中的作用  

The role of PKC to TNF-α induced β-1,4-galactosyltransferase-Ⅰ expression in endothelial cells

作　　者：季玉红[1] 贲志云[1] 汪晓莺[1] 孙晓雷[1] 钱佶[1] 肖锋[1] 

机构地区：[1]南通大学医学院免疫学与微生物学教研室

出　　处：《细胞与分子免疫学杂志》2010年第7期653-656,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30770488)

摘　　要：目的：研究蛋白激酶C（PKC）对肿瘤坏死因子-α（TNF—α）诱导内皮细胞β-1，4-半乳糖基转移酶-I（β-1，4．GaIT—I）表达的调节作用，以及对内皮细胞骨架结构改变及其黏附能力的影响。方法：分别用PKC激动剂或几种不同类型的PKC抑制剂预处理人脐静脉内皮细胞（HUVECs），再用TNF一仅刺激HUVECs，应用RT—PCR、Western blot方法检测B．1，4-GaIT—I表达变化，应用细胞荧光染色观察B-1，4-GalT—I催化的糖链的表达变化及细胞骨架结构的改变，通过内皮-单核细胞黏附试验观察HUVECs黏附能力的改变。结果：几种不同类型的PKC抑制剂均能不同程度的抑制TNF-α刺激HUVECs引起的β-1，4-GaIT—I表达的上调，PKC激动剂能够使上调的β-1，4-GaIT—I的表达进一步增加；在HUVECs中β-1，4-GaIT—I与细胞骨架有共同定位，PKC抑制剂显著抑制TNF—α诱导的内皮细胞骨架蛋白的重构和β-1，4-GaIT—I细胞内的再分布；PKC抑制剂显著抑制TNF—α诱导的内皮一单核细胞黏附能力的上调。结论：PKC可能参与调节TNF—α诱导的HUVECsβ—1，4-GaIT—I的表达，并且可能多种类型的PKC参与了这一调节过程；PKC可能通过对β-1，4-GaIT—I的调节进而影响炎症过程中内皮细胞骨架蛋白的重构及内皮细胞与单核细胞的黏附能力。AIM： To investigate the role of protein kinase C （PKC） to regulate the expression of β-1,4-galactosyltransferase-I （ β-1,4-GalT-I ） and the influence to cytoskeleton and adherence ability of human umbilical vein endothelial cells（HUVECs） when stimulated by tumor necrosis factor （TNF-α）. METHODS： Cultured HUVECs were pretreated by various PKC inhibitors or PMA, an excitomotor of PKC respectively for 30 minutes, then stimulated by TNF-α for 4 hours, 15-1,4-GalT-I expression were detected by RT- PCR and Western blot, expression of beta-1, 4-galactosylated carbohydrate chains and cytoskeleton were assayed by immumofluorescence, adherence ability of HUVECs was observed by endothelial-monocyte cell adherence test. RE- SULTS： Up-regulated expression of 15-1,4-GalT-I and beta- 1,4-galactosylated carbohydrate chains in HUVECs stimulated by TNF-a were suppressed by PKC inhibitors and increased by PMA. F-actin and β-1, 4-GalT-I were partly colocalized in HUVECs, PKC inhibitor inhibited the effect of TNF-α on the distribution of f-actin and β-1, 4-GalT-I. Adherence ability of HUVECs enhanced by LPS was significantly suppressed by PKC inhibitor. CONCLUSION： PKC signal transduction pathway may participate in regulating β- 1,4-GalT-I expression in endothelial cells （EC） stimulated by TNF-α, furthermore, polytypes of PKC may participate in this regulating process; PKC might regulate cytoskeleton reorganization and adherence ability of EC through β-1, 4- GalT-I during inflammation.

关 键 词：β-1 4-半乳糖基转移酶-I 蛋白激酶C 内皮细胞 TNF-Α 

分 类 号：R392.12[医药卫生—免疫学]