丹参乙酸镁对缺氧／复氧条件下心肌微血管内皮细胞的保护及作用机制

Protecctive effect and mechanism of mafnesium lithospermate B on hypoxia/reoxygenation-in-duced apoptosis in cardiac microvascular endothelial cells

目的探讨丹参乙酸镁（MLB）在抑制缺氧/复氧（H/R）诱导心肌微血管内皮细胞凋亡中的作用及机制。方法体外培养及建立大鼠心肌微血管内皮细胞H/R模型，MLB浓度分别为12.5、25、50、62.5 μmol/L，通过TUNEL法检测心肌微血管内皮细胞的凋亡率及MLB的最适浓度。确定MLB最适浓度后分为正常对照组、H/R组、H/R＋MLB组及H/R＋MLB＋锌原卟啉IX（ZnPPIX）组。采用Transwell法检测细胞迁移能力，流式细胞仪检测细胞凋亡率。结果与H/R组比较，不同浓度的MLB均可抑制细胞凋亡，并呈剂量依赖性，50 μmol/L MLB组出现明显保护作用（P〈0.01）。ZnPPIX干预后，细胞迁移能力明显减弱（P〈0.05），细胞凋亡率明显升高（P〈0.01）。结论丹参乙酸镁能够减少缺氧/复氧诱导的心肌微血管内皮细胞凋亡，其机制可能与上调血红素加氧酶（HO-1）水平有关。

ObjectiveTo investigate the role and possible mechanism of mafnesium lithospermate B （MLB） on hypoxia/reoxygenation-induced apoptosis of cardiac microvascular endothelial cells （CMECs）.MethodsCMECs of rat were cultured in vitro and the model of hypoxia/reoxygenation （H/R） was established. MLB with four concentrations （12.5, 25, 50, 62.5 μmol/L） was selected. TdT-mediated dUTP nick-end labeling （TUNEL） method was used to detect apoptosis index of CMECs, and determine the optimum concentration of MLB with lowest rate of apoptosis. Then CMECs were randomly divided into normal control group, H/R group, H/R ＋ MLB group and H/R＋ MLB＋ Zn-protoporphyrin IX （ZnPPIX） group. Cell migration ability was detected by transwell method and the apoptosis index was measured by flow cytometry.ResultsCompared with H/R group, different concentrations of MLB can inhibit apoptosis of CMECs in a dose-dependent menner, and there was a significant protective effect in 50 μmol/LMLB group （P〈0.01）. After the intervention of ZnPPIX, the cell migration ability was significantly decreased （P〈0.05）, and the apoptosis index was significantly increased （P〈0.01）.ConclusionsMLB can reduce apoptosis of CMECs induced by hypoxia/reoxygenation. Its mechanism may be related to the increase of heme oxygenase level.