球状脂联素在波动性高血糖诱导人脐静脉内皮细胞凋亡中的作用

EFFECT OF GLOBULAR ADIPONECTIN ON APOPTOSIS OF HUMAN UMBILICAL VEIN ENDOTHELIAL CELLS INDUCED BY INTERMITTENT HIGH GLUCOSE

目的球状脂联素在波动性高血糖诱导人脐静脉内皮细胞凋亡中的作用。方法在不同条件下体外培养人脐静脉内皮细胞,分别或联合加入球状脂联素(gAD)、单磷酸腺苷激活蛋白激酶(AMPK)的激活剂AICAR和AMPK的阻滞剂araA。采用MTT比色法测定细胞活性,流式细胞仪检测细胞凋亡率,Western blot检测AMPKα和磷酸化AMPKα蛋白表达。结果分别与对照组和恒定高血糖组比较,波动性高血糖显著抑制细胞活性和增加细胞凋亡率。gAD明显抑制波动性高血糖诱导的细胞凋亡。AICAR和gAD可明显激活AMPK的表达。araA可明显抑制gAD诱导的AMPK蛋白表达。结论波动性高血糖比恒定性高血糖更易促进内皮细胞凋亡,gAD明显抑制波动性高血糖诱导内皮细胞凋亡,其机制可能与激活AMPK有关。

Objective To explore the effect of globular adiponectin on the apoptosis of human umbilical vein endothelial cells induced by intermittent high glucose.Methods Human umbilical vein endothelial cells were cultured in different conditions.Cells were treated respectively or conjugately by globular adiponectin（gAD）,AICAR or araA.MTT assay was used to determine the cellular viability.Annexin Ⅴ-FITC/PI double staining was used to detected apoptosis.Western blot was used to detect protein expression levels of AMPKα and P-AMPKα.Results Compared with those of the control group or the permanent high glucose group,cellular viability decreased and apoptosis increased in the alternating high glucose group.3μg/ml gAD protected against cell apoptosis in the alternating high glucose group.AMPK was activated by gAD and AICAR respectively or both in the alternating high glucose group.But AMPK was inactivated when HUVECs were pretreated by araA.Conclusion Cellular apoptosis is more obvious in the intermittent high glucose environment than in the permanent high glucose environment.That gAD protects against cellular apoptosis induced by intermittent high glucose is associated with the activation of AMPK.