高糖增强过氧化氢诱导的牛主动脉内皮细胞凋亡。

High glucose enhances H2O2-induced apoptosis in bovine aortic endothelial cells.

作者信息

Guo X, Liu W L, Chen L W, Guo Z G

机构信息

Laboratory of Molecular Pharmacology, Hunan Medical University, Changsha 410078, China.

出版信息

Acta Pharmacol Sin. 2000 Jan;21(1):41-5.

PMID:11263246

Abstract

AIM

To investigate the effect of high glucose on hydroperoxide (H2O2)-induced apoptosis in cultured bovine aortic endothelial cells (BAEC).

METHODS

BAEC were cultured and passaged in normal glucose (5.5 mmol.L-1, NG) and high glucose (25 mmol.L-1, HG). Morphologic changes and quantification of apoptotic cells were determined under fluorescence microscope after H2O2-treated BAEC for 24 h with Hoechst 33258 staining. DNA fragmentation was visualized by agarose gel electrophoresis. The expression of phospho-p38 Ca(2+)-calmodulin dependent protein kinase (CCDPK, formerly called MAPK) was measured by Western blotting.

RESULTS

H2O2 elicited typical apoptotic morphologic changes (chromatic condensation, nucleus fragmentation). At 100 -300 mumol.L-1, both NG- and HG-BAEC incubated with H2O2 for 24 h increased cell apoptosis and phospho-p38 CCDPK expression in a concentration-dependent manner. In HG-BAEC, H2O2 induced DNA fragmentation at a lower concentration than that in NG-BAEC, and the apoptotic cell count in HG-BAEC was also higher than that of NG-BAEC (P < 0.05). Similarly, the expression of phospho-p38 CCDPK induced by H2O2 was up-regulated in HG-BAEC (P < 0.05).

CONCLUSION

High glucose enhances H2O2-induced apoptosis in BAEC, which is related to high expression of phospho-p38 CCDPK.

摘要

目的

研究高糖对过氧化氢（H2O2）诱导的培养牛主动脉内皮细胞（BAEC）凋亡的影响。

方法

将BAEC在正常葡萄糖（5.5 mmol·L-1，NG）和高糖（25 mmol·L-1，HG）条件下进行培养和传代。用Hoechst 33258染色处理H2O2作用24小时后的BAEC，在荧光显微镜下观察凋亡细胞的形态变化并进行定量分析。通过琼脂糖凝胶电泳观察DNA片段化情况。采用蛋白质免疫印迹法检测磷酸化p38钙调蛋白依赖性蛋白激酶（CCDPK，原称MAPK）的表达。

结果

H2O2引发典型的凋亡形态学变化（染色质凝聚、细胞核碎裂）。在100 - 300 μmol·L-1浓度下，NG - BAEC和HG - BAEC与H2O2共孵育24小时均使细胞凋亡和磷酸化p38 CCDPK表达呈浓度依赖性增加。在HG - BAEC中，H2O2诱导DNA片段化的浓度低于NG - BAEC，且HG - BAEC中的凋亡细胞计数也高于NG - BAEC（P < 0.05）。同样，H2O2诱导的HG - BAEC中磷酸化p38 CCDPK的表达上调（P < 0.05）。