中文摘要：

目的探讨P13-K／Akt、有丝分裂原活化蛋白激酶（mitogen—activatedproteinkinases，MAPKs）信号通路在氧化应激诱导人脐静脉内皮细胞凋亡中的作用。方法体外分离培养人脐静脉内皮细胞（HUVEC），采用不同浓度的过氧化氢（H2O2）刺激细胞建立氧化应激损伤的模型，采用M1Tr法检测细胞活力，用原位末端标记（TUNEL）法检测细胞凋亡，双氢罗丹明123（DHR）染色检测细胞内活性氧（ROS）水平，Westernblot检测P13-K／Akt和MAPKs信号通路关键蛋白的表达情况。结果各浓度H2O2组细胞活力（OD值）较对照组明显降低（P〈0．01），各浓度H2O2组不同作用时间点OD值差异无统计学意义（P〉0．05）；200μmol／LH2O2刺激HU-VECs24h后，细胞凋亡率及胞内ROS水平较对照组明显升高（P〈0．05）；H2O2组P—Akt、P—C—Jun、p—p38及p—ERKl／2蛋白表达较对照组明显增加（P〈0．05），而采用抗氧化剂白藜芦醇（RES）预处理2h后，这-作用被逆转。结论P13-K／Akt和MAPKs信号通路可能参与ROS介导的HUVEC细胞凋亡。

英文摘要：

Objective To explore the effects of PI3 - K/Akt and MAPKs signal pathway in oxidative stress induced HUVECs apoptosis. Methods Human umbilical vein endothelial cells （HUVECs） are isolated and cultured, then cells are exposed to different concentrations of hydrogen peroxide （H2O2 ） to establish the oxidative stress injury model. Ceils viability is evaluated by MTT assay. The apoptosis and intracellular ROS of HUVECs are detected by TUNEL and Rhodamine 123 staining though flow cytometry. Protein expressions of PI3 - K/Akt and MAPKs signal pathway are tested by Western blot. Results HUVECs viability （OD value ） were significantly lower in each H202 stimulation groups compared with control group （ P 〈 0.01 ）, and showed no concentration and time correlation （P 〉 0. 05 ）. 24 hours after 200 μmol/L H2O2 stimulation, the apoptosis index and intrcellular ROS level of HUVECs were significantly increased compared with control group （ P 〈 0.05 ）. The protein expressions of p - Akt, p - c - Jun, p - p38, p - ERK1/2 in H2O2 group were significantly up- regulated compared with control group （P 〈 0. 05 ）, and this effects were reversed when pre - incubating cells with antioxidant resveratrol （RES） for 2 h. Conclusion Our data suggest that PI3 - K/ Akt and MAPKs signal pathway are possibly involved in oxidative stress induced HUVECs apoptosis.