脱氧胆酸钠对人脐静脉内皮细胞凋亡的影响

Effect of Sodium Deoxycholate(SD) on Human Umbilical Vein Endothelial Cell Apoptosis

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 discuss the effect of sodium deoxycholate （SD）on the apoptosis of HUVECsin vitro. Methods： （1）After HUVECs were respectively exposed to different final concentrations of SD （0,0.015 mg/mL, 0.05 mg/mL, 0.15 mg/mL, 0.5 mg/mL,1.0 mg/mLcell viability was detected by CCK-8 assay and cell apoptosis was detected by TUNEL staining. （2）After 0.15 mg/mL SD was acted on HUVECs for 4 h, 6 h, 8 h, Western blot was used to detect the activation of Caspase-3, 7, 9 and PARP.（3）Observe the effect of Z-DEVD-FMK （a Caspase-3 inhibitor） on the groups with SD at a dose of 0.15 mg/mL. Results： CCK8 assay showed that with the increasing of SD concentrations（0 - 1.0 mg/mL）and action time（0 -12h）, cell viability of HUVECs decreased accordingly. Cell viability was 80% at a dose of 0.15 mg/mL, while merely less than 10% at a dose of 1.0 mg/mL.TUNEL staining displayed that the more the concentrations of SD, the more cell apoptosis there were. From Western blot, it could be seen that after adding SD into HUVECs the activation of Caspase-3, 7, 9 and PARP rose. Z-DEVD-FMK could inhibit the effect of SD at a dose of 0.15 mg/mL on the activation of PARP. Conclusion： SD could induce HUVEC apoptosis through the activation of Caspase Proteinases.