摘要
目的探讨3-脱氧葡糖醛酮（3-DG）对人血管内皮细胞（VECs）存活的影响及作用机制。方法从人脐静脉分离血管内皮细胞并与不同浓度的3-DG在体外共同培养。细胞调亡用形态学和原位末端标记（TUNEL）法证实，凋亡或死亡细胞数量用FITCAnnexin-V及碘化丙碇（PI）染色后，以流式细胞仪检测。细胞内氧化水平以氧化敏感的荧光染料2，7-二氢二氯荧光素（DCFH）染色，用流式细胞仪测定。结果3-DG刺激后，内皮细胞出现凋亡的形态学变化且TUNEL阳性，调亡与死亡细胞数量随3-DG作用时间和浓度的增加而增多；同时，3－DG刺激后细胞内氧化水平升高；抗氧化剂N-乙酰半胱氨酸和羰基化合物清除剂氨基胍抑制3－DG引起的细胞氧化应缴并减少细胞凋亡与死亡。结论3－DG能够诱导VECs凋亡，其作用机制可能是通过增加细胞内氧化应激。这些结果提示糖尿病和慢性肾功能衰竭循环3－DG蓄积可能是这类患者血管并发症的发生机制之一。
Objective To elucidate the effect of 3-deoxyglucosone (3-DG) on viability of human vascular endothelial cells(VECs). Methods VECs were isolated from human umbilical vein and cultured with s-DC in vitro. Apoptosis was observed by light microscope and TUNEL assay, and quantitated by flow cytomoter using FITC Annexin-V and propidium iodid (PI) staining. Intracellular oxidative levels were rneasured by flow cytometric assay using an oxidant sensitive dye-2, 7-dichlorefluoresin (DCFH). Results Apoptosis occured when VECs were incubated with 3-DG, evidenced by morphological changes and increased proportion of TUNEL positive cells. Apoptosis and death induced by 3-DG were both in a time- and dose-dependent manner. The levels of intracellular oxidation increased simultaneously. The effects of 3-DG on viability and oxidation of VECs were inhibited when a dicarbonyl scavanger aminogUanidine(AG) were included into the culture, or pre-incubated VECs with an oxidative inhibitor, N -acetylcysteine (NAC ).Conclusions 3-DG induces apoptosis of VECs by increasing intracellular oxidation level, which can he blocked by NAC and AG. These results indicate that accumulation of 3-DG in plasma may be involved in the pathogeneses of vascular diseases seen in diabetes and chronic renal failure.
出处
《中华肾脏病杂志》
CAS
CSCD
北大核心
2000年第2期76-79,共4页
Chinese Journal of Nephrology
基金
国家自然科学基金!39970341
