目的观察缺氧-复氧(H-R)损伤对大鼠心肌微血管内皮细胞(MMECs)PI3K、Akt和缺氧诱导因子-1α(HIF-1α)mRNA表达的影响。方法培养SD大鼠MMECs,以1×106/ml的密度接种于96孔板(100μl/孔)或直径6 cm培养皿(2 ml/皿),随机分为H-R组(缺氧2 h,复氧2 h)和正常对照组(N组)。Hoechst染色荧光显微镜覌察凋亡细胞形态,Annexin V-FITC和PI双标记法测定细胞凋亡率,MTT法测定细胞活力,RT-PCR检测PI3K、Akt、和HIF-1αmRNA转录水平。结果与N组比较,H-R组可见大量细胞坏死、脱落,MMECs活力降低,细胞凋亡率升高,HIF-1α、PI3K、Akt mRNA表达上调(P<0.05或P<0.01)。结论 H-R促使细胞凋亡,抑制细胞增殖,上调Akt和HIF-1αmRNA的表达,是MMECs H-R损伤机制之一。 Objective To observe the effects of hypoxia-reoxygenation (H-R) injury on the expression of PI3K, Akt and hypoxia inducible factor-1α (HIF-1α) mRNA in rat cardiac microvascular endothelial cells (MMECs) Methods SD rat MMECs were cultured and plated in 96-well plates (100 μl / well) or 6 cm diameter dishes (2 ml / dish) at a density of 1 × 106 / ml and were randomly divided into HR group (hypoxia 2 h, Oxygen 2 h) and normal control group (N group). Hoechst staining was used to observe the morphological changes of apoptotic cells. Annexin V-FITC and PI double labeling methods were used to determine the apoptotic rate. MTT assay was used to determine cell viability. The transcription levels of PI3K, Akt and HIF-1α mRNA were detected by RT-PCR. Results Compared with group N, a large number of cells were necrotic and detached in H-R group. The activity of MMECs was decreased and the apoptosis rate was increased. The expressions of HIF-1α, PI3K and Akt mRNA were up-regulated (P <0.05 or P <0.01). Conclusions H-R can induce cell apoptosis, inhibit cell proliferation and up-regulate the expression of Akt and HIF-1αmRNA, which is one of the mechanisms of H-R injury in MMECs.