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目的:观察高糖慢性处理对血管内皮细胞内质网应激标志蛋白葡萄糖调节蛋白78(GRP78)表达的影响及其作用机制。方法:培养的HUVECs细胞用含正常糖(5.5mmol/L)或高糖(30mmol/L)的培养基处理不同时间后,用MTT法测定细胞存活率,流式细胞仪测定细胞凋亡,Western blotting法测定蛋白表达情况。结果:与相应对照组相比,HUVECs细胞经高糖孵育24-48h后,GRP78蛋白表达量先增加后减少;而环加氧酶-2(COX-2)蛋白表达量逐渐增加。COX-2选择性抑制剂尼美舒利与高糖共孵育后,可明显抑制高糖引起的GRP78蛋白表达改变;同时尼美舒利可抑制高糖孵育48h后诱导的细胞凋亡。结论:高糖慢性处理可诱导HUVECs细胞GRP78表达先增加后减少,此过程依赖于COX-2蛋白的上调。
Objective: To observe the effect of chronic high glucose treatment on the expression of endoplasmic reticulum stress marker glucose regulated protein 78 (GRP78) in vascular endothelial cells and its mechanism. Methods: The cultured HUVECs were treated with normal glucose (5.5mmol / L) or high glucose (30mmol / L) for different time. The cell viability was measured by MTT assay. The apoptosis was measured by flow cytometry blotting method to determine protein expression. Results: Compared with the corresponding control group, the expression of GRP78 protein increased first and then decreased in HUVECs after being incubated with high glucose for 24-48 hours, while the expression of cyclooxygenase-2 (COX-2) protein gradually increased. COX-2 selective inhibitor nimesulide co-incubated with high glucose, can significantly inhibit GRP78 protein expression caused by high glucose changes; nimesulide at the same time inhibit the high glucose-induced apoptosis after 48h. CONCLUSION: Chronic high glucose treatment can induce the expression of GRP78 in HUVECs to increase first and then decrease, which depends on the up-regulation of COX-2 protein.