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SS-GAG对高糖状态下血管内皮细胞NO释放及ET-1分泌的影响

王瑞鑫1,王春波2,罗世滨1,李裕强1   

  1. 1. 山东大学威海分校海洋学院, 山东 威海 264209; 2. 青岛大学医学院, 山东 青岛 266071
  • 收稿日期:2007-12-11 修回日期:1900-01-01 出版日期:2008-04-16 发布日期:2008-04-16
  • 通讯作者: 李裕强

Effect of SS-GAG on ET1 and NO secretion of vascular endothelial cells damaged by highly concentrated glucose

WANG Rui-xin1,WANG Chun-bo2,LUO Shi-bin1,LI Yu-qiang1   

  1. 1. Marine College of Shandong University at Weihai, Weihai 264209, Shandong, China;2. Medical College of Qingdao University, Qingdao 266071, Shandong, China
  • Received:2007-12-11 Revised:1900-01-01 Online:2008-04-16 Published:2008-04-16
  • Contact: LI Yu-qiang

摘要: 目的观察高糖状态下扇贝裙边糖胺聚糖(scallop skirtglycosaminoglycan, SS-GAG)对体外培养的血管内皮细胞(ECV-304)一氧化氮(NO)释放及内皮素-1(ET-1)分泌的影响。方法在体外培养生长良好的ECV-304分别加入不同浓度葡萄糖,建立高糖损伤细胞模型,选择葡萄糖终浓度为55.5mmoL/L的培养液作为高糖环境,加入不同浓度SS-GAG继续培养至12、24、36、48h,倒置显微镜下观察细胞生长状态,特异性硝酸还原酶法检测细胞内NO,放射免疫法测定ET-1。结果① 高糖能抑制ECV-304细胞生长,细胞形态发生改变。 SS-GAG减轻高糖对ECV-304损伤,SS-GAG保护组细胞形态基本完好;② 高浓度葡萄糖促进ET-1的分泌,与正常对照比较差异有统计学意义(P<0.01)。而NO含量则随作用时间的延长表现出先升高后下降的动态变化过程,与正常对照比较差异有统计学意义(P<0.01)。SS-GAG对高糖状态下ECV-304细胞分泌ET1、NO具有调节作用,使其趋于正常,与正常对照比较差异无统计学意义(P>0.05),与高糖比较差异有统计学意义(P<0.01)。结论高浓度葡萄糖对ECV-304具有明显的损伤作用,可直接导致ECV-304形态和功能异常,引起ET-1分泌增多以及损伤内皮NO系统使NO分泌失调。SS-GAG可减轻高糖对ECV-304细胞的损伤,通过调节 ET和NO平衡,起到保护ECV-304作用。

Abstract: To study the effects of scallop skirt glycosaminoglycan (SS-GAG) on ET1 and NO secretion of ECV304 damaged by highly concentrated glucose. Methods The vascular endothelial cell ECV304 was cultured in vitro, and the damage mode1 of endothelia1 cells was established by highly concentrated glucose.Different concentrations of SS-GAG was mixed with cell culture fluid containing highly concentrated glucose (55.5mmol/L), and the cell morphology was observed after cultured for 12h, 24h, 36h and 48h, respectively. Nitric oxide (NO) inside cells were determined by the assay of nitric acid reductase. Radioimmunoassay was used to determine ET-1. ResultsCell morphology was changed and proliferation was depressed under the highly concentrated glucose, and a dose-dependent reaction occurred. SSGAG weakened the damage effects of highly concentrated glucose, and made these cells keep their normal morphology. The concentration of ET-1 in the cell culture supernatant was increased compared with the normal control group. Differences had statistics meaning compared with the normal control group (P<0.01). And the NO inside cells initially increased, and then reduced. Differences had statistics meaning compared with the normal control group (P<0.01). There were obvious differences between the SS-GAG group and the negative control group. With modulation of SS-GAG, ET-1 and NO inclined to the normal level,difference occurred between the SSGAG and the highly concentrated glucose groups. Difference had statistics meaning compared with the highly concentrated glucose group (P<0.01) but not with the normal control group (P>0.05). ConclusionsHighly concentrate glucose has obvious damaged effects on vascular endothelial cells, which may induce the cell morphology change, dysfunction, ET1 increase and NO secretion. SS-GAG will weaken this damage and protect the cells by modulating the balance of ET1 and NO.

Key words: Scallop, Glycosaminoglycan, Glucose, Vascular endothelial cell, Endothelin 1, Nitric oxide

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