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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 investigate the protective effect of glucagon-like peptide 1 (GLP-1) on cardiomyocyte apoptosis induced by advanced oxidized protein product (AOPPs) and its mechanism. Methods H9C2 cardiomyocytes were used as control. The cells were treated with AOPPs, AOPPs, GLP-1, AOPPs + GLP-1 and AOPPs + GLP-1 + LY294002 respectively for 24 h. Cell viability was detected by cck-8 method. DCFH-DA fluorescent probe was used to detect the intracellular reactive oxygen species (ROS) production. Annexin V-FITC / PI double staining was used to detect cell apoptosis. , Bcl-2, Bax, active-caspase-3 protein expression. Results Apoptosis of cells induced by AOPPs was significantly lower than that of AOPPs [(1.409 ± 0.099) vs (0.929 ± 0.083), P <0.01], and decreased the production of reactive oxygen species induced by AOPPs [(47.817 ± 0.878)% vs (25.413 ± 2.597 ), P <0.01] and apoptosis [(15.773 ± 3.130)% vs (9.715 ± 0.757)%, P <0.05]. GLP-1 restored the phosphorylation of Akt and Bad inhibited by AOPPs, Bcl-2, Bax, a pro-apoptotic protein, and active-caspase-3. Conclusions GLP-1 can protect AOPPs-induced apoptosis of H9C2 cardiomyocytes by changing the balance of pro-apoptotic and anti-apoptotic proteins through the PI3K / Akt / Bad signaling pathway.