【摘要】 目的探讨红景天苷对大鼠心肌微血管内皮细胞(CMECs)缺血/再灌注损伤的影响及其可能的机制。方法分离培养大鼠CMECs,建立模拟缺血/再灌注模型,分为对照组、模拟缺血/再灌注(SI/R)组、SI/R+红景天苷组(1.0、2.5、5.0、10.0μmol/L)组。待红景天苷最适浓度确认为5μmol/L后,增加SI/R+红景天苷+LY[磷酯酰肌醇3激酶(PI3k)特异性抑制剂LY294002]组。MTT法检测细胞增殖能力,细胞划痕实验检测细胞迁移能力,TUNEL法检测细胞凋亡,采用Western blot检测Akt磷酸化水平,以及凋亡抑制蛋白生存素和Bcl-2的表达。结果与对照组相比较,SI/R组CMECs增殖能力明显降低(0.410±0.011比0.200±0.014,P=0.041),凋亡率显著上升(4.15%±0.12%比26.05%±0.97%,P=0.018),而与SI/R组相比,SI/R+红景天苷组(1.0、2.5、5.0、10.0μmol/L)细胞增殖能力明显升高并呈剂量依赖性,细胞迁移率,而凋亡率则明显下降(均为P<0.05),LY294002组凋亡指数与SI/R+红景天苷组相比显著提高(22.03%±0.98%比16.28%±1.40%,P=0.029)。与SI/R组相比较,红景天苷可显著上调Akt的磷酸化,以及上调抗凋亡蛋白生存素和Bcl-2的表达(均为P<0.05),而此作用可被LY294002显著抑制(均为P<0.05)。结论红景天苷可显著抑制缺血/再灌注损伤诱导的CMECs凋亡,促进细胞存活,改善细胞功能,其作用机制可能与激活PI3K/Akt,以及上调凋亡抑制蛋白生存素和Bcl-2表达相关。更多还原

【Abstract】 Objective To explore the protective effect of salidroside against ischemia/reperfusion injury of cardiac microvascular endothelial cells( CMECs) and the underlying mechanisms. Methods CMECs isolated from the hearts of adult rats were divided to three groups: Control group,simulated ischemia / reperfusion( SI / R) group and simulated ischemia / reperfusion + Salidroside group. Then added with SI / R + Salidroside + LY( PI3 k specific inhibitor LY294002) group after the optimal dose of Salidroside( 5 μmol / L) was identified. The cell viability of CMECs was measured by MTT assay and migration ability of CMECs was detected by cell scratch wound assay. The apoptosis of CMECs was detected by TUNEL method. The phosphorylation of Akt and the expression the anti-apoptotic proteins survivin and Bcl-2 were analyzed by Western blot. Results Both of the cell viability and migration ability were impaired after SI / R( P < 0. 05 vs. control),and the apoptosis index was increased compared with control group( 26. 05% ±0. 97% vs. 4. 15% ± 0. 12%,P = 0. 018). While administration of salidroside during reperfusion dramatically attenuate the dysfunction of CMECs and the apoptosis induced by I / RI( all P < 0. 05). WB assay proved that both phosphorylation of Akt and the expression the anti-apoptotic proteins survivin and Bcl-2 were increased when compared with SI / R group and which were all inhibited when added with LY294002 I( all P <0. 05). Conclusions Salidroside has protective effect of CMECs against ischemia / reperfusion injury through activating Akt phosphorylation and might related with up-regulation of protein survivin and Bcl-2.更多还原