失神经骨骼肌NF-kappa B的表达变化与线粒体通透性转换孔开放及细胞凋亡的关系

The relationship between the change in NF-κB and mitochondrion permeability transition pore （MPTP） and apoptosis in denervated skeletal muscles of rats

目的 通过研究去神经骨骼肌细胞核转录因子kappa B（ntlcle．arfactor of kappa B，NF-kappa B）p65与线粒体通透性转换孔（mitochondrion permeability transition pore，MPTP）及细胞凋亡的关系，探讨失神经肌萎缩的相关机制。方法Wistar大鼠30只，随机分为健康对照组、去神经2d组、去神经7d组、去神经14d组、去神经28d组。建立右下肢腓肠肌失神经支配模型，对照组仅做假手术。采用Western Bloting检测大鼠腓肠肌NF-kappa B p65蛋白表达水平，共聚焦荧光显微镜观察MPTP的开放情况，脱氧核糖核苷酸转移酶介导的缺口末端标记（Tunnel）技术检测细胞凋亡情况，并结合肌湿重进行相关性分析。结果大鼠失神经支配后随着肌湿重比的下降，NF-kappa B p65蛋白、MPTP的开放以及骨骼肌细胞的凋亡率在各个时间点持续增加，且明显高于对照组，差异有统计学意义（P〈0．05），NF-kappa B与MPTP的开放（r=-0．896，P〈0．01）以及凋亡率（r=0．890，P〈0．01）密切相关，并且NF-kappaB和凋亡率的升高与肌湿重比呈负相关关系（r分别为-0．919、-0．924，P均〈0．01）。结论NF-kappa B在失神经肌萎缩中起重要作用，其作用机制与MVIP的开放及细胞凋亡有关。

Objective -To analyze the relationship-of nuclear factor of kappaB （NF-κB） p65 to mitoehondrion permeability transition pore （MPTP） and apoptosis during denervation atrophy of the gastroenemius muscle, and delineate their potential roles in skeletal muscle atrophy. Methods Thirty Wistar rots were randomly divided into 5 groups： control group, 2 day denervation group, 7 day denervation group, 14 day denervation group, and 28 day denervation group. The gastrocnemius muscle was denervated by transection of the sciatic nerve. Sham operation was done in the control group. Levels of NF-κB p65 protein and the opening of the mitochondrial permeability transition pore in the gastrocnemius muscle were detected respectively by Westem Bloting and eonfoeal fluorescence microscopy. The apoptotic cells in atrophic muscles were quantitated with Tune[ Muscle wet weight was also measure for comparison. Results At different time points after denervation, the levels of NF-zB p65, the opening of MPTP and apoptosis of gastrocnemlus were significantly higher than those in the normal group （ P 〈 0.05）. Furthermore NF-κB p65 had a positive correlation with the opening of MPTP （ r = - 0. 896, P 〈 0.01 ） and with the ratio of apoptusis （ r = 0. 890, P 〈 0.01 ）, while the increase of NF-κB p65 and apoptosis was negatively correlated with the ratio of muscle wet weight （ r = - 0.919, -0.924, P 〈0.01）. Conclusion NF-κB plays an important role in denervation skeletal muscle atrophy. This effect is related to the opening of MPTP and apoptesis.