Shi Kexin, Wang Xiaoxue, Ke Zhifei, Li Junping
School of Human Sports Science, Beijing Sport University, Beijing, China.
Key Laboratory of Sports and Physical Health of Ministry of Education, Beijing Sport University, Beijing, China.
J Muscle Res Cell Motil. 2023 Dec;44(4):311-323. doi: 10.1007/s10974-023-09660-6. Epub 2023 Oct 27.
This study aimed to explore the occurrence of necroptosis in skeletal muscle after eccentric exercise and investigate the role and possible mechanisms of ZBP1 and its related pathway proteins in the process, providing a theoretical basis for the study of exercise-induced skeletal muscle injury and recovery. Forty-eight male adult Sprague-Dawley rats were randomly divided into a control group (C, n = 8) and an exercise group (E, n = 40). The exercise group was further divided into 0 h (E0), 12 h (E12), 24 h (E24), 48 h (E48), and 72 h (E72) after exercise, with 8 rats in each subgroup. At each time point, gastrocnemius muscle was collected under general anesthesia. The expression levels of ZBP1 and its related pathway proteins were assessed using Western blot analysis. The colocalization of pathway proteins was examined using immunofluorescence staining. After 48 h of eccentric exercise, the expression of necroptosis marker protein MLKL reached its peak (P < 0.01), and the protein levels of ZBP1, RIPK3, and HMGB1 also peaked (P < 0.01). At 48 h post high-load eccentric exercise, there was a significant increase in colocalization of ZBP1/RIPK3 pathway proteins, reaching a peak (P < 0.01). (1) Eccentric exercise induced necroptosis in skeletal muscle, with MLKL, p-MLKL, and HMGB1 significantly elevated, especially at 48 h after exercise. (2) After eccentric exercise, the ZBP1/RIPK3-related pathway proteins ZBP1, RIPK3, and p-RIPK3 were significantly elevated, particularly at 48 h after exercise. (3) Following high-load eccentric exercise, there was a significant increase in the colocalization of ZBP1/RIPK3 pathway proteins, with a particularly pronounced elevation observed at 48 h post-exercise.
本研究旨在探讨离心运动后骨骼肌中坏死性凋亡的发生情况，并研究ZBP1及其相关通路蛋白在此过程中的作用和可能机制，为运动性骨骼肌损伤与恢复的研究提供理论依据。将48只成年雄性Sprague-Dawley大鼠随机分为对照组（C，n = 8）和运动组（E，n = 40）。运动组进一步分为运动后0小时（E0）、12小时（E12）、24小时（E24）、48小时（E48）和72小时（E72），每个亚组8只大鼠。在每个时间点，在全身麻醉下采集腓肠肌。采用蛋白质免疫印迹分析评估ZBP1及其相关通路蛋白的表达水平。采用免疫荧光染色检测通路蛋白的共定位。离心运动48小时后，坏死性凋亡标记蛋白MLKL的表达达到峰值（P < 0.01），ZBP1、RIPK3和HMGB1的蛋白水平也达到峰值（P < 0.01）。高负荷离心运动后48小时，ZBP1/RIPK3通路蛋白的共定位显著增加，达到峰值（P < 0.01）。（1）离心运动诱导骨骼肌发生坏死性凋亡，MLKL、p-MLKL和HMGB1显著升高，尤其是在运动后48小时。（2）离心运动后，ZBP1/RIPK3相关通路蛋白ZBP1、RIPK3和p-RIPK3显著升高，特别是在运动后48小时。（3）高负荷离心运动后，ZBP1/RIPK3通路蛋白的共定位显著增加，运动后48小时升高尤为明显。
