糖基化终末产物诱导心肌细胞老化的机制  

Mechanisms of advanced glycation end products induced cardiomyocytes aging

作　　者：李世玲 郭聪娴 王俊宏 郭妍 

机构地区：[1]南京医科大学第一附属医院老年心血管科,江苏南京210029 [2]江苏盛泽医院心血管科,江苏苏州215228

出　　处：《南京医科大学学报（自然科学版）》2015年第8期1066-1071,共6页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省科技厅自然科学基金(BK2011382);江苏省"六大人才高峰"(2011WSN-029);江苏省卫生厅科技项目(Z201301)

摘　　要：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 study the mechanisms of cardiomyocytes aging induced by advanced glycation end products. Methods:Neonatal rat cardiac myocytes were cultured with AGEs,anti-RAGE antibody for 48 h. Senescence-associated beta galactosidase(SA-β-Gal) activity was observed via SA-β-Gal assay in neonatal rat cardiomyocytes. p53,p16,LC3 and Beclin1 were measured by Western blot. We observed mitochondrial membrane potential and the generation of reactive oxygen species(ROS) by JC-1 and DCFH-DA methods. Results:After intervention for 48 h with AGEs,SA-β-Gal activity and the expression of p53(P < 0.01),p16(P <0.01),LC3(P < 0.01) and Beclin1(P < 0.01) were significantly increased in the AGEs group compared to that of the control group,accompanied by the significantly decrease of mitochondrial membrane potential(P < 0.01) and remarkably increase of ROS(P < 0.01).After treated with anti-RAGE antibody,SA-β-Gal activity and the level of p53(P < 0.01),p16(P < 0.01),LC3(P < 0.05)and Beclin1(P < 0.01)were remarkably decreased compared with the AGEs group. However,inhibition of RAGE with anti-RAGE antibody remarkably increased mitochondrial membrane potential(P < 0.01) and significantly decreased ROS compared with AGEs group(P <0.01).Conclusion:AGEs-RAGE may induce aging of cardiomyocytes by mitochondrial damage,the generation of ROS and autophagy.

关 键 词：糖基化终末产物 糖基化终末产物受体 心肌细胞老化 线粒体 氧化应激 自噬 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]