CAJ | 학술논문

目的研究游离脂肪酸(FFA)水平升高导致的内皮一氧化氮合酶(eNOS)活性降低是否与蛋白激酶C(PKC)通路激活有关.方法传代培养的人脐静脉内皮细胞分为4组:对照组,油酸组(10、50、100、150、200 μmol/L),PKC抑制剂(GFX)组,油酸+GFX组.分别检测各组eNOS活性,检测对照组和油酸组PKC的表达和活性,以及各组eNOS磷酸化(p-eNOS)的表达.结果与对照组相比,油酸组eNOS活性呈剂量依赖性降低,p-eNOS(1177)的表达降低(0.0854±0.017 vs. 0.0134±0.023, P<0.05);PKC的表达出现转位,由胞浆转向胞膜,胞浆表达活性降低,胞膜表达活性增高,总活性增加.加入GFX后对正常内皮细胞eNOS活性无明显影响,但能部分改善油酸所致的内皮细胞eNOS活性的降低.结论油酸可损害内皮细胞eNOS的表达及磷酸化,其机理与PKC通路的激活有关.
목적 연구유리지방산(FFA)수평승고도치적내피일양화담합매(eNOS)활성강저시부여단백격매C(PKC)통로격활유관.방법 전대배양적인제정맥내피세포분위4조:대조조,유산조(10、50、100、150、200 μmol/L),PKC억제제(GFX)조,유산+GFX조.분별검측각조eNOS활성,검측대조조화유산조PKC적표체화활성,이급각조eNOS린산화(p-eNOS)적표체.결과 여대조조상비,유산조eNOS활성정제량의뢰성강저,p-eNOS(1177)적표체강저(0.0854±0.017 vs. 0.0134±0.023, P<0.05);PKC적표체출현전위,유포장전향포막,포장표체활성강저,포막표체활성증고,총활성증가.가입GFX후대정상내피세포eNOS활성무명현영향,단능부분개선유산소치적내피세포eNOS활성적강저.결론 유산가손해내피세포eNOS적표체급린산화,기궤리여PKC통로적격활유관.
Objective To observe whether the impairment of endothelial nitric oxide synthase (eNOS) activity by elevated level of free fatty acid (FFA) is associated with activation of protein kinase C (PKC). Methods The cultured human umbilical vein endothelial cells were used for four groups: control group, oleic acid(OA) group (10, 50, 100, 150, 200 μmol/L), GFX group and OA+GFX group. The activity of eNOS in each group was examined, and the activity and expression of PKC were determined in both control and OA groups. The method of immunocytochemistry was performed to detect the expression of p-eNOS(1177) in control, OA (100 μmol/L), GFX and OA+GFX groups respectively. Results Compared with control group, the eNOS activity of endothelial cells in OA group was significantly reduced in a dose-dependent manner, and the expression of p-eNOS(1177) were also impaired (0.0854±0.017 vs. 0.0134±0.023, P<0.05). PKC transposition from endochylema to cytomembrane was observed by confocal microscopy. The PKC activity was degraded in endochylema, while enhanced in cell membrane inversely, and the gross activity of PKC increased after all. When GFX, an inhibition of PKC, was added to endothelial cells at the present of OA or not, the impaired eNOS activity were partly improved. Conclusion Elevated level of FFA (here is oleic acid) leading to endothelial dysfunction results from a loss of eNOS activity and its serine phosphorylation, and it may be related to activation of PKC.