CAJ | 학술논문

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、10.0 umol/L）组。待红景天苷最适浓度确认为5 umol/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、10.0 umol/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。
목적：탐토홍경천감대대서심기미혈관내피세포( CMECs）결혈/재관주손상적영향급기가능적궤제。방법분리배양대서CMECs,건립모의결혈/재관주모형,분위대조조、모의결혈/재관주(SI/R）조、SI/R+홍경천감조(1.0、2.5、5.0、10.0 umol/L）조。대홍경천감최괄농도학인위5 umol/L후,증가SI/R+홍경천감+LY[린지선기순3격매(PI3k）특이성억제제LY294002]조。 MTT법검측세포증식능력,세포화흔실험검측세포천이능력,TUNEL법검측세포조망,채용Western blot검측Akt린산화수평,이급조망억제단백생존소화Bcl-2적표체。결과여대조조상비교, SI/R조CMECs증식능력명현강저(0.410±0.011비0.200±0.014,p=0.041）,조망솔현저상승(4.15%±0.12%비26.05%±0.97%, p=0.018）,이여SI/R조상비, SI/R+홍경천감조(1.0、2.5、5.0、10.0 umol/L）세포증식능력명현승고병정제량의뢰성,세포천이솔,이조망솔칙명현하강(균위 p <0.05）,LY294002조조망지수여SI/R+홍경천감조상비현저제고(22.03%±0.98%비16.28%±1.40%,p=0.029）。여SI/R조상비교,홍경천감가현저상조Akt적린산화,이급상조항조망단백생존소화Bcl-2적표체(균위p<0.05）,이차작용가피LY294002현저억제(균위p<0.05）。결론홍경천감가현저억제결혈/재관주손상유도적CMECs조망,촉진세포존활,개선세포공능,기작용궤제가능여격활PI3K/Akt,이급상조조망억제단백생존소화Bcl-2표체상관。
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 ( PI3k specific inhibitor LY294002 ) group after the optimal dose of Salidroside ( 5 umol/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.