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目的:研究老龄大鼠线粒体PT孔开放随年龄增长改变的机制。方法:采用酶解法分离大鼠心肌细胞,密度梯度离心法分离大鼠心肌线粒体,通过荧光法检测大鼠心肌线粒体中活性氧(ROS)含量,通过蛋白免疫印迹法检测腺甘酸转位酶(ANT)1、ANT2和亲和环素D(CyP-D)蛋白表达。结果:随月龄的增加,线粒体ROS生成逐渐增多,15月龄以上各组大鼠变化显著,同3月龄大鼠比差异具有统计学差异(P<0.01);同3月龄大鼠相比,18月龄大鼠线粒体ANT1和CyP-D表达增强,而ANT2表达减少(均P<0.05)。结论:ROS生成增多及ANT1、ANT2和CyP-D表达改变可能参与了随年龄增长线粒体PT孔开放改变的机制。
Objective: To investigate the mechanism of the opening of mitochondrial PT pore in aged rats with age. Methods: Myocardial cells were isolated by enzymatic method. Mitochondria were isolated by density gradient centrifugation. The content of reactive oxygen species (ROS) in myocardial mitochondria was detected by fluorescence spectrophotometry. Apoptosis was detected by Western blotting ) 1, ANT2 and Cyclophilin D (CyP-D) protein expression. Results: With the increase of age, the generation of mitochondrial ROS gradually increased. The rats in 15-month-old and above groups changed significantly. The difference with the 3-month-old rats was statistically significant (P <0.01) Compared with the control group, the expression of ANT1 and CyP-D in mitochondria of 18-month-old rats increased and the expression of ANT2 decreased (all P <0.05). CONCLUSIONS: Increased production of ROS and altered expression of ANT1, ANT2 and CyP-D may be involved in the mechanism of mitochondrial PT pore opening changes with age.