右归饮含药血清促进人血管内皮细胞增殖及抗氧化损伤的作用研究

Study of Proliferative and Antioxidant Effects of Youguiyin Containing Serum on Human Vascular Endothelial Cells

[目的]观察右归饮含药血清促进人血管内皮细胞增殖及对氧化损伤的保护作用,探讨右归饮防治激素性股骨头坏死的作用机理。[方法]取对数生长期的人血管内皮细胞,分为空白组、模型组、药物Ⅰ组、药物Ⅱ组。模型组和药物I组以600μM H2O2作用2h建立氧化损伤模型,空白组、药物II组照常培养,2h后药物I组和药物II组分别加入相同浓度右归饮含药血清,空白组和模型组则分别加入等量空白血清。各组干预24h后通过倒置相差数字成像显微镜观察细胞形态及密度、MTT比色法检测细胞OD值,并分别于24h、48h、72h通过Realtime-PCR测定空白组、模型组、药物Ⅰ组细胞中血管内皮细胞生长抑制因子(vascular endothelial growth inhibitor,VEGI)的表达水平。[结果]与空白组相比,药物Ⅱ组细胞密度显著增高,形态完整,且OD值高于空白组(P<0.001);与模型组相比,用右归饮含药血清干预后,药物Ⅰ组细胞密度增高,形态较为完整,OD值高于模型组(P<0.01);PCR结果显示药物Ⅰ组细胞的VEGI表达水平低于模型组(24h,48h,P<0.01)。[结论]右归饮含药血清能在一定程度上促进体外培养的人血管内皮细胞增殖,同时对细胞氧化损伤模型具有保护作用,其作用可能与下调VEGI基因表达有关。

[Objective]To study the prohferative and antioxidant effects of Youguiyin containing serum on human vascular endothehal ceils, and clarify its relevant mechanism treating steroid-induced necrosis of femoral head. [Method]When human vascular endothelial cells reached the logarithmic phase, they were divided into blank group, model group, drug group I and drug group II. The model group and drug group I were respectively treated with blank serum and Youguiyin containing serum, following 2h H2O2 pretreatment, while the normal group and drug group II respectively treated with blank serum and Youguiyin containing serum directly after 2h normal incubation. After 24h observation, all groups were analyzed by inversion digital imaging microscope for cell morphology and density and by MTT for cell viability. Real time PCR was apphed to test the gene expression of vascular endothehal growth inhibitor, VEGI after 24h, 48h and 72h. [Result]The drug group II showed morphological intact and remarkable increase of cell density compared with the blank group, and its MTT optical density was significantly higher than the blank group（P〈0.01）. Compared with the model group, the drug group I showed increased morphological intact arid cell density as well as increased MTT optical density（P〈0.01）. The real time PCR, results at 24h and 48h showed that VEGI expression in the drug group I was significantly lower than the model group（P〈0.01）, while the model group lower than the blank group（P〈0.01）. [Conclusion]Youguiyin containing serum can promote the prohferation of human vascular endothelial cells to a certain extent, and have protective effect to vascular oxidative damage. Such effects have relevance to the VEGI modulation.