摘要
目的:研究糖基化终产物(AGEP)对主动脉平滑肌细胞信号转导环节之一二酰基甘油(Dia)的影响和维生素E、氨基胍对Dia水平的干预作用.方法:采用放射酶标记、薄层层析、放射自显影技术测定细胞中Dia含量.结果:AGEP-BSA孵育的平滑肌细胞Dia水平在15 s和10min出现两个峰值,对照组和AGEP-BSA 50,11,200 mg·L^(-1)作用SMC的Dia水平分别为(220 ±15)pmol^(-1),(328±15)pmol·L^(-1),(662±33)pmol·L^(-1)和(940±43)pmol·L^(-1). Dia水平也随AGEP-BSA糖基化的程度和修饰BSA的糖浓度增加而增高.维生素E和氨基胍可明显减弱AGEP刺激平滑肌细胞Dia水平升高的效应.结论:AGEP-BSA能升高培养的大鼠血管平滑肌细胞中的Dia水平,维生素E和氨基胍能明显抑制这一效应.
AIM: To investigate whether diacylglycerol (Dia) signaling pathway is stimulated by advanced glycosylation end products (AGEP) and to determine effect of vitamin E and aminoguanidine on Dia level induced by AGEP in cultured rat aortic vascular smooth muscle cells (VSMC). METHODS: The effect of AGEP on Dia level in cultured VSMC was measured by radio-enzymatic assay. Quantitative measurements of [32P]phosphatidic acid were performed by thin-layer chromatography and autoradiography. RESULTS: The Dia level in VSMC incubated with AGEP-BSA was markedly increased in a concentration-dependent, biphasic manner. The early phase was rapid and transient, peaking at 15 s. The late phase reached the maximal level at 10 min and decayed slowly. The Dia levels in VSMC exposed to different concentrations of AGEP-BSA and AGEP-BSA glycosylated for various periods were greatly increased compared with control group. Vitamin E 50, 100 nmol·L-1 prevented the AGEP-BSA-stimulated elevation of Dia level in VSMC, from (940±43) pmol·L-1 to (599±38), (290±21) pmol·L-1, respectively. In aminoguani-dine-treated AGEP-BSA groups, Dia level in the cells incubated with the same concentration of AGEP-BSA (100, 200 mg·L-1) were decreased to (260±8) and(289±10) pmol·L-1, respectively. Early glycosylated low-density lipoproteins (LDL) did not affect Dia level. CONCLUSION: AGEP causes a robust stimulation of Dia signaling pathway in VSMC. Vitamin E and aminoguanidine attenuate the production of Dia.
出处
《中国药理学报》
CSCD
1999年第7期617-622,共6页
Acta Pharmacologica Sinica
基金
Project supported by the National Natural Science Foundation of China, № 39470314
关键词
糖基化终产物
甘油二酯类
血管平滑肌
advanced glycosylation end products
diglycerides
vascular smooth muscle
cultured cells
atherosclerosis
signal transduction
vitamin E
guanidines
phosphatidic acids
autoradiography
