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目的:研究糖基化终末产物(advanced glycation end products,AGEs)诱导心肌细胞老化的机制。方法 :AEGs、抗AGE受体(RAGE)抗体干预乳鼠心肌细胞48 h。通过Western blot和β-半乳糖苷酶(SA-β-Gal)染色分别检测老化相关蛋白p53、p16的表达及SA-β-Gal活性;采用JC-1、DCFH-DA分别测定线粒体膜电位、细胞内活性氧(reactive oxygen species,ROS);通过Western blot检测自噬相关蛋白LC3、Beclin1。结果:AGEs干预细胞48 h后与对照组相比,AGEs组SA-β-Gal活性明显增高,p53、p16的表达量明显增加(P<0.01),同时伴随线粒体膜电位的降低(P<0.01)及细胞内ROS水平增加(P<0.01),LC3与Beclin1表达量明显增加(P均<0.01);给予抗RAGE抗体干预后与AGEs组相比,SA-β-Gal活性降低,p53、p16的表达量明显降低(P<0.01),线粒体膜电位增高(P<0.01),细胞内ROS减少(P<0.01),LC3与Beclin1表达量降低(P<0.05和P<0.01)。结论:AGEs与其受体RAGE作用可能通过氧化应激及线粒体损伤与自噬诱导心肌细胞老化。
Objective: To investigate the mechanism of advanced glycation end products (AGEs) induced cardiomyocyte aging. Methods: AEGs and anti-AGE receptor (RAGE) antibodies were used to interfere neonatal rat cardiomyocytes for 48 h. The expression of p53, p16 and the activity of SA-β-Gal were detected by Western blot and SA-β-Gal staining. The mitochondrial membrane potential, Reactive oxygen species (ROS) was detected by Western blot. The autophagy-related proteins LC3 and Beclin1 were detected by Western blot. Results: Compared with the control group, the activity of SA-β-Gal in AGEs group increased significantly (P <0.01) and the mitochondrial membrane potential decreased (P <0.01) (P <0.01). The expression levels of LC3 and Beclin1 were significantly increased (all P <0.01). Compared with AGEs group, the activity of SA-β-Gal and the expression of p53 and p16 (P <0.01). The mitochondrial membrane potential was increased (P <0.01), the intracellular ROS was decreased (P <0.01), and the expression of LC3 and Beclin1 was decreased (P <0.05 and P <0.01). Conclusion: The effect of AGEs and its receptor RAGE may induce cardiomyocyte aging through oxidative stress and mitochondrial damage and autophagy.