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目的观察芪丹通脉片(QDTMT)对缺氧内皮细胞的保护机制。方法原代培养、鉴定人脐静脉内皮细胞(HuVECs)。制备含QDTMT的血清并干预传代的HuVECs。将HuVECs根据不同处理因素随机分为3组:空白对照组、生理盐水组和QDTMT组,用流式细胞仪检测各组细胞的凋亡率。用免疫荧光染色法检测生理盐水组和QDTMT组细胞上血管内皮生长因子受体2(VEGFR2)的表达。结果培养的HuVECs中具有特征性的短棒状细胞器Weible-Palade小体。流式细胞仪检测结果显示,与空白对照组相比,缺氧培养的HuVECs的凋亡率增加,但QDTMT组内皮细胞的凋亡率明显低于生理盐水组(P<0.05)。免疫荧光染色检测可见QDTMT组内皮细胞VEG-FR2的表达强于生理盐水组。结论QDTMT抗缺氧的HuVECs凋亡的作用可能与其促进VEGFR2的表达有关。
Objective To observe the protective mechanism of Qi Dan Tong Mai tablets (QDTMT) on hypoxic endothelial cells. Methods Primary culture, identification of human umbilical vein endothelial cells (HuVECs). Serum containing QDTMT was prepared and interfered with passaged HuVECs. The HuVECs were randomly divided into three groups according to different treatment factors: blank control group, saline group and QDTMT group. The apoptosis rate of each group was detected by flow cytometry. Immunofluorescence staining was used to detect the expression of vascular endothelial growth factor receptor 2 (VEGFR2) in the normal saline group and the QDTMT group. RESULTS: Wells-Palade bodies characterized by short rod-shaped organelles were found in cultured HuVECs. The results of flow cytometry showed that compared with the blank control group, the apoptosis rate of HuVECs in hypoxia increased, but the apoptosis rate of endothelial cells in QDTMT group was significantly lower than that in saline group (P <0.05). Immunofluorescence staining showed that the expression of VEG-FR2 in endothelial cells of QDTMT group was stronger than that of saline group. Conclusion The anti-hypoxia effect of QDTMT on apoptosis of HuVECs may be related to its effect on the expression of VEGFR2.