Cytoskeletal vimentin protein expression in rats with exhaustive eccentric exercise injury

doi:10.3969/j.issn.2095-4344.2014.18.016

Abstract

Abstract:

BACKGROUND: Due to different experimental methods, the change of cytoskeleton proteins after centrifugal movement is still controversial.
OBJECTIVE: To establish exhaustive eccentric exercise injury model in rats and to observe cytoskeletal vimentin protein expression at different time.
METHODS: A total of 48 male Sprague-Dawley rats were randomly and equally divided into six groups: quiet control group, immediately after exercise group, and 12, 24, 48, 72 hours after exercise groups. In the exercise groups, the rats were subject treadmill exercise at the speed of 16 m/min in a -16° slope, for 100 minutes at the interval of 5 minutes. The quiet control group maintained unchanged, without exercise. The cytoskeletal vimentin was detected with immunohistochemical staining using anti-vimentin antibody, and vimentin expression changes were observed at different time after exhaustive eccentric exercise through the changes of target area percentage.
RESULTS AND CONCLUSION: The results of target area percentage showed that, there was no significant difference between quiet control group and immediately after exercise group (P > 0.05). Compared with the immediately after exercise group, the target area percentage was slightly increased in 12 hours after exercise group, without significant difference (P > 0.05). Compared with the 12 hours after exercise group, the target area
percentage was also slightly increased in 24 hours after exercise group, without significant difference (P > 0.05). Compared with quiet control group and immediately after exercise group, the target area percentage was increased in 24 hours exercise group (P < 0.05). Compared with immediately and 12 hours after exercise groups, the target area percentage was increased significantly in 48 hours after exercise group (P < 0.01). Compared with 48 hours after exercise group, the target area percentage began to decline in 72 hours after exercise group (P < 0.05), which was higher than quiet control level. After exhaustive eccentric exercise, cytoskeletal vimentin begin to express at varying degrees. The vimentin expression is increased gradually at 12 hours and reached the peak at 48 hours, then begins to decrease.

中国组织工程研究杂志出版内容重点：肾移植；肝移植；移植；心脏移植；组织移植；皮肤移植；皮瓣移植；血管移植；器官移植；组织工程

全文链接：

Key words: athletic injuries, vimentin, muscle, skeletal, sports medicine

CLC Number:

R318

Liu Xiang-dong, Li Yang. Cytoskeletal vimentin protein expression in rats with exhaustive eccentric exercise injury[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(18): 2880-2885.

Figures/Tables 4

运动后12 h组与运动后即刻组相比，超微结构进一步损伤。可见肌纤维排列紊乱、扭曲，肌纤维间隙增宽，明暗带模糊。Z线局部紊乱、扭曲。横小管肿胀。细胞膜有改变，局部溶解，肌细胞核肿胀，固缩，形状不规则。线粒体空泡变性，部分嵴缺损(图1C)。运动后24 h组超微结构的损伤程度更加明显。可见肌纤维排列不规则、紊乱、扭曲，肌丝间肿胀，明暗带模糊，无法区分A、I带。Z线排列不规则，可见Z线流，Z线呈锯齿状，Z线局部消失。肌细胞膜固缩、部分溶解，肌细胞核浓缩、核膜模糊。线粒体多见空泡变性、肿胀、嵴部分缺损、紊乱(图1D)。运动后48 h组较运动后24 h组超微结构的损伤程度有所减轻。可见肌纤维间肌丝水肿。Z线局部错位，可见Z线流变。肌细胞膜溶解、皱缩，核膜肿胀、模糊。线粒体絮状物改变，外膜局部破坏，嵴部分缺损，常见空泡变性(图1E)。运动后72 h组大鼠骨骼肌超微结构损伤程度明显减轻。可见肌纤维排列接近正常，未见肌纤维水肿。肌细胞膜部分皱缩，能够分清明暗带和M线。多数线粒体比较完整，部分线粒体嵴缺损、空泡变性，絮状改变逐渐变化到髓样改变(图1F)。 2.3 大鼠骨骼肌波形蛋白免疫组化染色结果本实验用抗波形蛋白抗体对大鼠骨骼肌波形蛋白进行免疫组化染色，通过观察其目标面积百分比的变化反映波形蛋白的相对含量(图2)。 2.4 一次力竭性离心运动后大鼠骨骼肌波形蛋白表达的定量结果结果显示，安静对照组、运动后即刻组、运动后12 h组、运动后24 h组、运动后48 h组和运动后72 h组的目标面积百分比分别为(26.92±4.62)%，(27.24±7.19)%，(28.62±9.42)%，(31.89±7.07)%，(39.18±8.70)%，(34.47± 7.91)%。安静对照组和运动后即刻组两组间差异无显著性意义(P > 0.05)，提示大鼠在经过一次力竭性离心运动后即刻，波形蛋白表达与安静组大鼠波形蛋白表达无差别，说明大鼠一次力竭性离心运动后即刻，骨骼肌细胞并没有出现明显的损伤。与运动后即刻组相比，运动后12 h组目标面积百分比略有增加，但差异无显著性意义(P > 0.05)；与运动后12 h组相比，运动后24 h组目标面积百分比略有增加，但差异无显著性意义(P > 0.05)；与安静对照组和运动后即刻组相比，运动后24 h组目标面积百分比有所增加 (P < 0.05)；与运动后即刻组和运动后12 h组相比，运动后 48 h组目标面积百分比明显增加(P < 0.01)；与运动后48 h组相比，运动后72 h组目标面积百分比有所下降(P < 0.05)，但没有恢复到安静时水平。结果提示目标面积百分比在运动后12 h逐渐增加，以运动后48 h组达最高，而后开始减少。说明在一次力竭性离心运动后不同时刻，大鼠骨骼肌中波形蛋白出现不同程度的表达(表1，图3)。"

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