Objectives To study the effects of contusion and exhaustive exercise on gene expression of MG53, PTRF, Pax7 and β-catenin in skeletal muscle of rats, and reveal the repair mechanism of skeletal muscle injury. Methods Forty-two male Wistar rats were randomly divided into 7 groups, with 6 rats in each group. All groups were euthanized at different time points after exhaustive exercise and contusion, respectively, while the control group was euthanized in resting state. The right gastrocnemius muscles were measured for mRNAs of MG53, PTRF, Pax7 and β-catenin by real time PCR. Results MG53 mRNA and PTRF mRNA of skeletal muscle in groups immediately after exhaustive exercise and after contusion increased significantly (p<0.05), while the two indices decreased constantly at 24 and 48 hours after injury with a similar change trend. Compared with the control group, Pax7 mRNA of skeletal muscle as a marker showed no significant difference in exhaustive exercise groups, but decreased at 48 hours after contusion (p<0.05). β-catenin mRNA of skeletal muscle down-regulated significantly over 24 hours after injury, then activated with an increased value at 48 hours after contusion (p<0.05). As a whole, the variations in the above indices in the contusion groups covered a wider range than in the exhaustive exercise groups. Conclusion The cytomembrane repair mechanism of MG53 and PTRF began immediately after the end of exhaustive exercise and contusion. Activation of Pax7 as the satellite cell marker took longer, and Wnt/β-catenin pathway showed first a decrease and then an increase resulting from the time-dependent gene expression during the repair of skeletal muscle injury. Level of evidence III, Therapeutic studies investigating the results of treatment.
Objective To study the effects of contusion and exhaustive exercise on the expression of degradation-related factors MuRF1 and MAFbx in the skeletal muscle of rats and describe the repair mechanism of skeletal muscle injury. Methods Forty-two male SD rats were randomly divided into 7 groups. The rats in each group were killed at different time points (0h, 24h, 48h) after exhaustive exercise (E0, E24, E48) and contusion (D0, D24, D48), respectively, and in the resting state in control group (C). The right gastrocnemius muscles were resected and divided into two parts, one for the mRNAs of MuRF1 and MAFbx by real-time PCR, and the other for protein measurement by Western blotting. Results Compared with the control group, the MuRF1 mRNA and protein expression of the skeletal muscle in the E0 group was markedly increased (P <0.05) and followed by a downward trend in E24 the E48 groups. On the other hand, MuRF1 mRNA expression of the skeletal muscle in the D24 group was significantly upregulated (P <0.01), then decreased in the D48 group (P <0.01). Meanwhile, compared with the C group, MAFbx mRNA gene expression continued to be upregulated in D24 and D48 (P <0.05), but decreased in E24 and E48 (p<0.01). On the other hand, the NF-κB protein contents of the skeletal muscle in the D0, D24, and D48 groups, as well as in the E48 group, were markedly downregulated (P <0.05), and the one in E48 was also remarkably downregulated (P <0.05). Conclusion NF-κB may negatively regulate the process of protein degradation by the NF-κB / MuRF1 signal pathway. Level of evidence III; Therapeutic studies investigating the results of treatment.
Objective: To study the effects of exhaustive exercise and contusion on autophagy-related factors Beclin1, LC3 and PINK1 expression in the skeletal muscle of rats. Methods: Forty-two male SD rats were randomly divided into 7 groups, 6 rats in each group: C, D0, D24, D48, E0, E24, and E48. Each group of rats was killed and dissected at the different respective time points specified above. The whole quadriceps femoris of the left hind limbs were removed and divided into two parts, one for mRNAs of Beclin1, LC3 and PINK1 by real-time fluorescent quantitative PCR, and the other for LC3 protein by Western blotting. Results: Compared with group C, the contents of Beclin1 mRNA, PINK1 mRNA, and LC3 mRNA in the immediate exhaustive exercise group (E0) were significantly reduced p<0.01. However, the levels of PINK1 mRNA, LC3 mRNA, and LC3 protein in skeletal muscle cells increased significantly in the 48 hours after exhaustion (E48) p<0.05, suggesting that cell autophagy had an increasing trend during the recovery period. Meanwhile, compared with the C group, the contents of Beclin1 mRNA, PINK1 mRNA, and LC3 mRNA in the immediate blunt contusion group (D0) increased significantly p<0.01 and were followed by a downward trend. Conclusion: Generally, there were differences between the blunt contusion and exhausted exercise models at each recovery phase. The gene expression of the autophagy-related factors was not high in the early exhaustive exercise recovery phase and subsequently followed an upward trend. But the above factors increased significantly in the immediate and early recovery phases after blunt contusion. Injury from blunt contusion may be more severe than exhaustive exercise-induced-injury, so the autophagy starts earlier according to the changes in autophagy-related factors. Level of evidence III; Therapeutic studies investigating the results of treatment.
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