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目的:探讨梓醇对高糖诱导的人脐静脉内皮细胞(HUVECs)凋亡的保护作用及其机制。方法:用33 mmol/L葡萄糖作用于人脐静脉内皮细胞24 h建立凋亡模型,分别以终浓度为20、100、500μg/ml的梓醇进行保护,MTT比色法检测细胞存活率;硫代巴比妥酸法检测细胞上清液中MDA的水平;流式细胞术检测细胞凋亡率;RT-PCR检测ET-1基因的表达情况;Western Blot法检测NF-κB信号通路相关蛋白的表达情况。结果:与正常组相比,33 mmol/L葡萄糖作用人血管内皮细胞24 h后细胞存活率明显降低;细胞上清液中MDA水平、细胞凋亡率明显升高;ET-1基因、p65蛋白的表达明显上调;IκB-α蛋白的表达则明显下调。与凋亡组相比,20、100、500μg/ml的梓醇保护组细胞存活率明显升高,细胞上清液中的MDA水平、细胞凋亡率均明显降低;ET-1基因、p65蛋白的表达明显下调;IκB-α蛋白的表达则明显上调,并且呈浓度依赖性。结论:梓醇对高糖诱导的人脐静脉内皮细胞凋亡具有保护作用,其机制可能与梓醇降低了血管内皮细胞的IκB-α的磷酸化水平,从而抑制了NF-κB信号通路的激活有关。
Objective: To investigate the protective effect of catalpol on apoptosis of human umbilical vein endothelial cells (HUVECs) induced by high glucose and its mechanism. Methods: Apoptosis models were established by treating human umbilical vein endothelial cells with 33 mmol / L glucose for 24 h. The apoptosis rate was determined by catalpol at final concentrations of 20, 100 and 500 μg / ml, respectively. Cell viability was measured by MTT assay. The level of MDA in the cell supernatant was detected by substituting barbituric acid method; the apoptosis rate was detected by flow cytometry; the expression of ET-1 gene was detected by RT-PCR; the expression of NF-κB signal pathway related protein Express the situation. Results: Compared with the normal group, the survival rate of human vascular endothelial cells decreased significantly after treated with 33 mmol / L glucose for 24 h. The levels of MDA and apoptosis in the supernatant of the cells were significantly increased. The expressions of ET-1, p65 The expression of IκB-α protein was significantly down-regulated. Compared with the apoptotic group, the cell viability was significantly increased in the 20, 100 and 500μg / ml catalpol-protective groups, while the levels of MDA and apoptosis in the cell supernatant were significantly decreased. The levels of ET-1, p65 The expression of IκB-α protein was significantly up-regulated, and in a concentration-dependent manner. CONCLUSION: Catalpol has a protective effect on the apoptosis of human umbilical vein endothelial cells induced by high glucose, and its mechanism may be related to catalpol reducing the phosphorylation of IκB-α in vascular endothelial cells and inhibiting the activation of NF-κB signaling pathway related.