摘要
【目的】探讨NOX4信号通路在胰高血糖素样肽-1(GLP-1)拮抗糖基化终末产物(AGE)诱导的内皮细胞氧化损伤的作用机制。【方法】实验组分为对照组、AGE组、AGE+GLP-1组、AGE+NOX4 si RNA组及AGE+阴性si RNA组,RTPCR检测NOX4 m RNA表达,Western Blotting技术检测NOX4蛋白表达情况,流式细胞仪检测细胞凋亡率、ROS生成水平。【结果】与对照组相比,AGE可诱导细胞NOX4蛋白表达(P=0.001)及NOX4 m RNA表达(P<0.05)明显增高;加入GLP-1后,增高的NOX4蛋白(P=0.003)及NOX4 m RNA(P<0.05)明显下降。与AGE组相比,NOX4 si RNA可抑制AGE诱导的ROS生成(P=0.011)、细胞凋亡(P<0.05);阴性si RNA对AGE诱导的ROS生成(P=0.958)、细胞凋亡(P=0.169)无明显影响。【结论】GLP-1至少部分通过抑制NOX4蛋白表达拮抗AGE诱导的人脐静脉内皮细胞氧化损伤。
[Objective] To discuss the role of NOX4 signaling pathway in the antagonistic mechanism of Glucagon-like Peptide-1 against the oxidative damage on vascular endothelial cells induced by advanced glycation endproducts. [ Method ] Experimental group was divided into a control group, AGE group, AGE + GLP-1 group, AGE +NOX4 siRNA group and AGE + negative control siRNA group. The expression of NOX4 mRNA was detected by RT-PCR. The expression of NOX4 protein was detected by Western blotting. The apoptosis rate and the levels of ROS were detected by flow cytometry. [Results] Compared with control group, AGE significantly promoted the expression of NOX4 protein (P = 0.001 ) and NOX4 mRNA (P 〈 0.05). After added GLP-1 into the AGE group, it significantly inhibited the expression of NOX4 protein (P = 0.003 ) and NOX4 mRNA (P 〈 0.05). Compared with AGE group, NOX4 siRNA inhibited the ROS generation (P = 0.011 ), the apoptosis of endothelial cells (P 〈 0.05). Negative control RNA had no influence on the ROS generation (P = 0.958), the apoptosis of endothelial cells (P = 0.169). [ Conclusion] GLP-1 antagonize oxidative damage on human umbilical endothelial cells induced by advanced glycation endproducts at least partly through inhibiting the expression of NOX4 protein.
出处
《中山大学学报(医学科学版)》
CAS
CSCD
北大核心
2016年第2期197-201,209,共6页
Journal of Sun Yat-Sen University:Medical Sciences
基金
广东省自然科学基金(S2011010002074)
清远市科技计划(2011B011112044)