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目的:探讨脱氧胆酸钠(SD)对体外培养的人脐静脉内皮细胞(HUVEC)凋亡的影响。方法:(1)以不同终浓度(0、0.015mg/mL、0.05 mg/mL、0.15 mg/mL、0.5 mg/mL、1.0 mg/mL)的脱氧胆酸钠分别作用于人脐静脉血管内皮细胞,使用CCK-8检测细胞活力、TUNEL荧光染色检测细胞凋亡;(2)以终浓度为0.15 mg/mL的SD作用于HUVEC4、8、12 h后用Western blot检测Caspase-3、7、9蛋白及PARP活化情况;(3)观察Caspase-3抑制剂Z-DEVD-FMK对0.15 mg/mL脱氧胆酸钠组的影响。结果:CCK8结果显示随SD浓度(0~1.0 mg/mL)及作用时间(0~12 h)增加,HUVEC活力降低,0.15 mg/mL时活力为80%,1.0 mg/mL时细胞活力仅不到10%;Tunel检测示随着SD浓度的增加HUVEC凋亡明显增多;Western Blot结果示SD作用于HUVEC后Caspase-3、7、9蛋白及PARP活化明显增加;Z-DEVD-FMK明显抑制了0.15 mg/mLSD引起的PARP活化。结论:脱氧胆酸钠(SD)通过启动Caspase级联反应介导了人脐静脉内皮细胞的凋亡。
Objective: To investigate the effect of sodium deoxycholate (SD) on the apoptosis of cultured human umbilical vein endothelial cells (HUVECs) in vitro. Methods: (1) Sodium deoxycholate with different final concentrations (0,0.015 mg / mL, 0.05 mg / mL, 0.15 mg / mL, 0.5 mg / mL and 1.0 mg / mL) Cell viability was detected by CCK-8, and apoptosis was detected by TUNEL staining. (2) HUVECs were treated with SD at a final concentration of 0.15 mg / mL for 8, 8 and 12 h, then Western blot was used to detect Caspase-3, 9 protein and PARP activation; (3) To observe the effect of Caspase-3 inhibitor Z-DEVD-FMK on 0.15 mg / mL sodium deoxycholate group. Results: The results of CCK8 showed that the viability of HUVEC decreased with the increase of SD concentration (0-1.0 mg / mL) and the action time (0-12 h), and the viability of HUVEC was 80% at 0.15 mg / mL. To 10%; Tunel test showed that HUVEC apoptosis increased significantly with the increase of SD concentration; Western Blot results showed that the activation of Caspase-3, 7, 9 protein and PARP increased significantly after treated with SD; Z-DEVD-FMK significantly inhibited PARP activation induced by 0.15 mg / mL SD. Conclusion: Sodium deoxycholate (SD) mediates the apoptosis of human umbilical vein endothelial cells by activating Caspase cascade.