胰升血糖素样肽1对晚期氧化蛋白产物诱导的心肌细胞凋亡保护作用的研究

Protective role of GLP-1 on AOPPs-induced apoptosis of cardiomyocytes

目的探讨胰升血糖素样肽1(GLP-1)对晚期氧化蛋白产物(AOPPs)诱导的心肌细胞凋亡的保护作用及机制。方法以H9C2心肌细胞为研究对象,用空白对照、RSA对照、AOPPs、GLP-1、AOPPs+GLP-1及AOPPs+GLP-1+LY294002分别干预细胞24 h。采用cck-8法检测细胞的增殖活力DCFH-DA荧光探针检测细胞内活性氧的产生;Annexin V-FITC/PI双染色法检测细胞凋亡;Western blot检测细胞内p-Akt、p-Bad、Bcl-2、Bax、active-caspase-3蛋白的表达。结果 GLP-1抵抗AOPPs引起的细胞增殖活力下降[(1.409±0.099)vs(0.929±0.083),P<0.01],减少AOPPs诱导的活性氧簇产生[(47.817±0.878)%vs(25.413±2.597)%,P<0.01]及细胞凋亡[(15.773±3.130)%vs(9.715±0.757)%,P<0.05];GLP-1能恢复被AOPPs抑制的Akt及Bad的磷酸化,增加细胞抗凋亡蛋白Bcl-2、减少促凋亡蛋白Bax及凋亡执行子active-caspase-3的表达。结论 GLP-1通过PI3K/Akt/Bad信号通路改变促凋亡及抗凋亡蛋白间的平衡,保护AOPPs诱导的H9C2心肌细胞凋亡。

Objective To explore the protective role and mechanisms of glucagon-like peptide-1 （GLP-1） on advanced oxidation protein products （AOPPs）-induced apoptosis of cardiomyocytes. Methods The H9C2 cells were selected in this study and divided into blank control group, RSA control group,and groups treated with indicated concentrations of AOPPs with or without GLP-1, and AOPPs ＋ GLP-1＋LY294002 for 24 hours respectively. Cell viability was detected by CCK-8 assay. The ROS level was detected by DCFH DA fluorescent probe. The cell apoptosis was tested by fluorescent staining and flow cytometry. The expression of p-Akt, p-Bad, Bcl-2, Bax, and active-caspase-3 proteins were evaluated by Western blot. Results GLP-1 attenuated AOPPs induced cytotoxicity[（0. 929±0. 083） vs （1. 409±0. 099）,P〈0. 01],decreased AOPPs induced ROSE（47. 817±0. 878）% vs （25. 413±2. 597）% ,P〈0. 01] and apoptosis[（15. 773 ± 3. 130） % vs （9. 715 ± 0. 757） %, P〈0.01]. GLP-1 improved AOPPs-induced phosphorylation of Akt and Bad,increased the expression of Bcl- 2, and decreased the expression of Bax and the activation of caspase-3. Conclusion GLP-1 protects cardiomyocytes against AOPP-induced apoptosis,predominantly via the PI3K/Akt/Bad pathway.