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目的:探讨大鼠肢体缺血-再灌注损伤骨骼肌钙调蛋白表达的变化。方法:30只SD大鼠随机分为对照组(control组)、缺血-再灌注组(IR组)。对照组所有大鼠用橡皮筋松弛环绕右后肢不阻断血流;IR组按张连元所报道的方法复制SD大鼠右后肢缺血-再灌注模型。各组动物于相应时间点在右后肢相应区域,切取约1000mg腓肠肌,冰盐水快速洗去血液,滤纸吸干水份,剪碎制成组织匀浆,用于RyR1和SERCA1的RT-PCR检测。结果:①骨骼肌型RyR1mRNA的RT-PCR检测结果:在肢体缺血-再灌注损伤再灌注8h内,IR组骨骼肌型RyR1mRNA在再灌注0h、2h表达上调,与对照组相比,其差异有统计学意义(P<0.01),4h时有所恢复,但8h时表达下调,与对照组相比,其差异有统计学意义(P<0.01)。②SERCA1mRNA的RT-PCR检测结果:对于SERCA1mRNA的表达,再灌注8h内随时间的延长其表达逐渐下降,与对照组相比,其差异均有统计学意义(P<0.01)。结论:在再灌注8h内,肢体缺血-再灌注损伤骨骼肌中RyR1mRNA早期表达上调,后期表达下降,SERCA1mRNA表达随时间的延长而逐渐下降。这可能是肢体缺血-再灌注骨骼肌损伤的重要原因之一。
Objective: To investigate the changes of skeletal muscle calmodulin (ICG) expression after limb ischemia-reperfusion injury in rats. Methods: Thirty SD rats were randomly divided into control group and ischemia-reperfusion group (IR group). All the rats in the control group did not block the blood flow around the right hind limbs with the rubber band relaxed. In the IR group, the right hindlimb ischemia-reperfusion model was reproduced by the method reported by Zhang Liangyuan. About 1000mg gastrocnemius muscles were taken from the corresponding regions of the right hind limbs at corresponding time points. The blood was quickly washed away with ice water and the filter paper was used to dry the water. The tissues were homogenized and cut for RT-PCR detection of RyR1 and SERCA1. Results: ①RT-PCR results of skeletal muscle RyR1mRNA: Within 8h after ischemia-reperfusion injury, the skeletal muscle RyR1mRNA in IR group was up-regulated at 0h and 2h after reperfusion, compared with the control group (P <0.01), restored at 4h, but decreased at 8h. The difference was statistically significant (P <0.01) compared with the control group. (2) RT-PCR results of SERCA1mRNA: The expression of SERCA1mRNA decreased gradually with the prolongation of time within 8h after reperfusion, and the difference was statistically significant (P <0.01) compared with the control group. Conclusion: Within 8 hours after reperfusion, the expression of RyR1mRNA in skeletal muscle after ischemia-reperfusion injury was up-regulated at the early stage and decreased at the late stage. The expression of SERCA1mRNA decreased gradually with the prolongation of time. This may be one of the major causes of limb ischemia-reperfusion skeletal muscle injury.