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^S358, 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^S232 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坏死性凋亡的发生情况，并探讨ZBP1及其相关通路蛋白在此过程中的作用及可能机制，为运动所致骨骼肌损伤及恢复的研究提供理论基础。 48 只雄性成年 Sprague-Dawley 大鼠被随机分为对照组（C，n = 8）和运动组（E，n = 40）。运动组又分为运动后0 h（E0）、12 h（E12）、24 h（E24）、48 h（E48）、72 h（E72）组，每组8只。在每个时间点，在全身麻醉下收集腓肠肌。使用蛋白质印迹分析评估 ZBP1 及其相关途径蛋白的表达水平。使用免疫荧光染色检查途径蛋白的共定位。离心运动 48 h 后，坏死性凋亡标志蛋白 MLKL 的表达量达到峰值（ P < 0.01），ZBP1、RIPK3、HMGB1 的蛋白水平也达到峰值（ P < 0.01）。高负荷离心运动后 48 h，ZBP1/RIPK3 通路蛋白的共定位显着增加，达到峰值（ P < 0.01）。 (1)离心运动引起骨骼肌坏死性凋亡，MLKL、 p -MLKL ^S358和HMGB1显着升高，特别是在运动后48 h。 (2)离心运动后，ZBP1/RIPK3相关通路蛋白ZBP1、RIPK3和p -RIPK3 ^S232显着升高，特别是在运动后48 h。 (3)高负荷离心运动后，ZBP1/RIPK3通路蛋白的共定位显着增加，运动后48小时观察到特别明显的升高。