心肌线粒体自噬相关蛋白BNIP3在运动预适应晚期保护效应中的变化

发布时间：2018-06-13 19:42

本文选题：BNIP3 + LC3　；参考：《上海体育学院》2017年硕士论文

【摘要】：研究目的:运动预适应(exercise preconditioning,EP)能够通过运动诱导机体产生内源性心肌保护效应,其机制涉及到很多方面。其中线粒体是EP心脏保护中重要的参与者,线粒体自噬的上调,可以及时清除受损线粒体,有利于维持细胞生存。而Bcl-2 19-k Da相互作用蛋白3(Bcl-2 19-k Da interacting protein 3,BN IP3)具有调控细胞自噬的功能,能与受损线粒体结合参与线粒体自噬。因此,本研究通过检测和观察心肌线粒体自噬相关蛋白BN IP3在EP晚期保护效应中的表达变化,探讨EP晚期保护效应和心肌线粒体自噬的关系。研究方法:SD大鼠随机分为对照组(C组)、力竭运动组(EE组)、晚期运动预适应组(LEP组)、晚期运动预适应+力竭运动组(LEP+EE组)和wortmannin+晚期运动预适应+力竭运动组(W+LEP+EE组)。在EP动物模型的基础上,用力竭运动致大鼠急性心肌损伤,用血浆c TnI含量和HBFP染色综合评价心肌损伤和保护的程度,用免疫荧光双标法检测大鼠心肌线粒体BNIP3和TOM20的共定位程度,用免疫印迹法检测大鼠心肌组织线粒体BN IP3和LC 3的表达变化。研究结果:与C组相比,EE组血浆c TnI和MOD值显著升高,BNIP3转位线粒体程度、BNIP3水平和LC3II/I水平显著升高;LEP组BNIP3转位线粒体程度和BNIP3水平显著升高,血浆c TnI、MOD值和LC3II/I水平无显著性差异。与EE组相比,LEP+EE组血浆c TnI和MOD值显著降低,BNIP3转位线粒体程度、BNIP3水平和LC3II/I水平显著降低。与LEP+EE组相比,W+LEP+EE组血浆c TnI和MO D值显著升高,BN IP3转位线粒体程度和BN IP3水平显著升高,LC3II/I水平无显著性差异。研究结论:EP晚期保护期可以减轻一次大强度力竭运动对心肌所造成的运动性损伤,细胞自噬部分参与EP晚期保护效应。一次大强度力竭运动和晚期EP均会导致BNIP3表达升高,参与诱导心肌细胞线粒体自噬。在EP晚期心肌保护效应中,BNIP3诱导的线粒体自噬参与了EP对运动性心肌损伤的保护作用。
[Abstract]:Objective: exercise preconditioning can induce endogenous myocardial protection through exercise, and its mechanism involves many aspects. Mitochondria is an important participant in EP heart protection. The upregulation of mitochondrial autophagy can clear damaged mitochondria in time and help to maintain cell survival. Bcl-2 19-k Da interacting protein 3n BN IP3) has the function of regulating autophagy, and can bind with damaged mitochondria to participate in mitochondrial autophagy. Therefore, the relationship between late EP protective effect and myocardial mitochondrial autophagy was studied by detecting and observing the expression of myocardial mitochondrial autophagy related protein BN IP3 in the late stage of EP. Methods Twenty SD rats were randomly divided into three groups: control group (C), exhaustive exercise group (EE group), advanced exercise preconditioning group (Lemp group), late exercise preconditioning exhaustive exercise group (Lemp EE group) and wortmannin advanced exercise preconditioning exhaustive exercise group (WLEP EE group). On the basis of EP animal model, acute myocardial injury induced by exhaustive exercise was used to evaluate the degree of myocardial injury and protection by plasma c TNI content and HBFP staining. The co-localization of BNIP3 and TOM20 in rat myocardial mitochondria was detected by immunofluorescence double labeling method, and the expression of BNIP3 and LC3 in myocardial mitochondria was detected by Western blot. Results: compared with group C, the levels of c TnI and MOD in plasma of EE group were significantly higher than those in group C, and the levels of BNIP3 and LC3II- / I were significantly higher than those of group C, but there was no significant difference between plasma c TnI mod value and LC3IIP I level in LEP group (P < 0.05). The level of BNIP3 translocation mitochondria and BNIP3 level in LEP group were significantly higher than those in group C. There was no significant difference in the level of plasma c TnI mod and LC3IIP I. Compared with EE group, plasma c TnI and MOD levels in LEP EE group were significantly lower than those in EE group. Compared with the LEP EE group, the plasma c TnI and MO D values in the WLEP EE group were significantly higher than those in the LEP EE group. There was no significant difference in the degree of mitochondrial translocation of BN IP3 and the level of BN IP3. There was no significant difference in the level of LC3II / I between the two groups. Conclusion during the late stage of protection, the myocardial injury caused by one intensive exhaustive exercise can be alleviated, and autophagy partly participates in the late protective effect of EP. Both a high intensity exhaustive exercise and late EP could increase the expression of BNIP3 and participate in the induction of mitochondrial autophagy in cardiomyocytes. Mitochondrial autophagy induced by BNIP3 is involved in the protective effect of EP on exercise induced myocardial injury.
【学位授予单位】：上海体育学院
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R87

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