葡萄籽原花青素对糖基化终产物损伤内皮细胞的保护作用

Protective effects of grape seed proanthocyanidins on endothelial cells intervened by advanced glycosylation end products

目的:研究不同浓度葡萄籽原花青素(GSPC)对糖基化终产物(AGE)作用下人脐静脉内皮细胞的保护作用及其机制。方法:体外培养人脐静脉内皮细胞(HUVEC),糖孵育法制备糖基化终产物修饰牛血清白蛋白(AGE-BSA)。实验分为6组,即空白对照组、实验对照组(BSA组)、损伤组(AGE组)、损伤加入GSPC低、中、高浓度组。将200 mg/L AGE作用于不同浓度GSPC培养4 h的内皮细胞,继续培养24 h,以细胞生存活力、血管性假血友病因子(vWF)、一氧化氮(NO)为检测指标。结果:200 mg/L AGE抑制HUVEC生存活力,细胞增殖活力下降为正常组的90.53%,GSPC预孵育组细胞生存活力逐渐增加,分别为正常组的0.95、1.12、1.23倍;AGE组vWF生成量较正常对照组明显增加(P<0.01),NO含量较正常对照组明显降低(P<0.01)。GSPC预孵育组可显著降低增高的vWF水平,NO生成量显著高于AGE组(P<0.01),100 mg/L GSPC预处理组NO水平恢复至正常水平。结论:AGE会损伤内皮细胞,抑制内皮细胞的生存活力,减少NO的生成。GSPC可抑制AGE对内皮细胞的损伤作用,且可抑制AGE减少NO生成的作用,并呈浓度依赖性,提示GSPC保护内皮细胞免受AGE损伤的机制可能与增加内皮细胞NO生成量有关。

Objective：To investigate the protective effect of grape seed proanthocyanidins （GSPC） on human umbilical vein endothelial cells （HUVEC） injured by advanced glycosylation end products （AGE） and its possible mechanism. Methods： AGE-modified bovine serum albumin （AGE-BSA） was prepared by incubating the BSA with a high concentration of glucose. The cultured HUVECs were divided into six groups ： the control group, the BSA group, the AGE group, the low dose GSPC ＋ AGE group, the middle dose GSPC ＋ AGE group, and the high dose GSPC ＋ AGE group. The cell proliferation of HUVEC, the Von Willebrand factor （vWF） and nitric oxide （NO） were determined. Results： After treatment with AGE, the proliferation of HUVEC was significantly reduced. The proliferation rate was 90.53% in the control group, while the proliferations of cells pretreated with GSPC of different concentrations were 0.95-fold, 1.12-fold, and 1.23-fold that of the control group. The vWF level in AGE-treated HUVEC was higher than that of the control group （P 〈0.01）. Pre-incubation with GSPC of different concentrations could decrease the vWF level increased by AGE in a dose-dependent manner. The NO content in the AGE-treated group was significantly lower than that of the control group （P 〈0.01）, while the AGE-decreased NO level was significantly increased in the GSPC-pretreated groups. Conclusion-AGE can inhibit HUVEC proliferation, injure HUVEC and decreased the NO content. GSPC could protect HUVEC against damage induced by AGE and increase the NO level in the AGE-exposed HUVEC. It is possible that the increased NO plays a key role in protecting the GSPC against damage induced by AGE.