CAJ | 학술논문

目的研究蛋白激酶c(PKC)对脂多糖(LPS)诱导内皮细胞β-1,4-半乳糖基转移酶-Ⅰ(β-1,4-galactosyltransferase-Ⅰ,β-1,4-GalT-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)30 min,LPS刺激HUVECs 4 h后,RT-PCR、Western blot方法检测β-1,4一GalT-Ⅰ表达变化,细胞荧光染色观察β-1,4-GalT-Ⅰ催化的糖链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地抑制LPS刺激HUVECs引起的β-1,4-GalT-Ⅰ表达的上调,PKC激动剂能够使上调的β-1,4-GalT-Ⅰ的表达进一步增加;在HUVECs中β-1,4-GalT-Ⅰ与细胞骨架有共同定位,PKC抑制剂显著抑制LPS诱导的内皮细胞骨架蛋白的重构和β-1,4-GalT-Ⅰ细胞内的再分布;PKC抑制剂显著抑制LPS诱导的内皮-单核细胞黏附能力的上调.结论 PKC可能参与调节LPS诱导的HUVECs β-1,4-GalT-Ⅰ的表达,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-GalT-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.
목적 연구단백격매c(PKC)대지다당(LPS)유도내피세포β-1,4-반유당기전이매-Ⅰ(β-1,4-galactosyltransferase-Ⅰ,β-1,4-GalT-Ⅰ)표체적조절작용,이급대내피세포골가결구개변급기점부능력적영향.방법 분별용PKC격동제혹궤충불동류형적PKC억제제예처리인제정맥내피세포(HUVECs)30 min,LPS자격HUVECs 4 h후,RT-PCR、Western blot방법 검측β-1,4일GalT-Ⅰ표체변화,세포형광염색관찰β-1,4-GalT-Ⅰ최화적당련표체변화급세포골가결구적개변,통과내피-단핵세포점부시험관찰HUVECs점부능력적개변.결과 궤충불동류형적PKC억제제균능불동정도지억제LPS자격HUVECs인기적β-1,4-GalT-Ⅰ표체적상조,PKC격동제능구사상조적β-1,4-GalT-Ⅰ적표체진일보증가;재HUVECs중β-1,4-GalT-Ⅰ여세포골가유공동정위,PKC억제제현저억제LPS유도적내피세포골가단백적중구화β-1,4-GalT-Ⅰ세포내적재분포;PKC억제제현저억제LPS유도적내피-단핵세포점부능력적상조.결론 PKC가능삼여조절LPS유도적HUVECs β-1,4-GalT-Ⅰ적표체,가능다충류형적PKC삼여료저일조절과정;PKC가능통과대β-1,4-GalT-Ⅰ적조절진이영향염증과정중내피세포골가단백적중구급내피세포여단핵세포적점부능력.
Objective To investigate the regulation of protein kinase C(PKC) to the expression of β-1,4-galactusyhransferase- Ⅰ ( β-1,4-GalT- Ⅰ ) and the influence on cytoskeleton and adherence ability of human umbilical vein endothelial cells(HUVECs) when stimulated by lipopolysaccharide (LPS). Methods Cultured HUVECs were pretreated by various PKC inhibitors or phorbol 12-myristate 13-acctate( PMA), an excitomotor of PKC respectively for 30 min, then stimulated by LPS for 4 h. β-1,4-GalT-Ⅰ expression were detected by RT-PCR and Western blot, expression of β-1,4-galactosylated carbohydrate chains and cytoskeleton were assayed by immumofluorescence, and adherence ability of HUVECs was observed by endothelialmonocyte cell adherence test. Results Up-regulated expression of β-1,4-GalT- Ⅰ and β-1,4-galactosylated carbohydrate chains in HUVECs stimulated by LPS were suppressed by PKC inhibitors and increased by PMA. F-actin and β-1,4-GalT- Ⅰ were partly co-localized in HUVECs. PKC inhibitor inhibited the effect of LPS 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 stimulated by LPS. 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.