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Ouabain exacerbates botulinum neurotoxin-induced muscle paralysis via progression of muscle atrophy in mice

机译:哇巴因通过小鼠肌肉萎缩的恶化加剧肉毒杆菌神经毒素诱导的肌肉麻痹

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摘要

Botulinum neurotoxin serotype A (BoNT/A) inhibits acetylcholine release at the neuromuscular junction in isolated muscles, and ouabain can partially block its effect. However, it is not clear whether ouabain attenuates BoNT/A-induced neuromuscular paralysis in vivo. In this work, we investigated the effects of ouabain on BoNT/A-induced neuromuscular paralysis in mice. Ouabain was administered to mice intraperitoneally immediately after a single injection of BoNT/A into skeletal muscle. The effects of ouabain on BoNT/A-induced muscle paralysis were assessed by quantitative monitoring of muscle tension and digit abduction via the digit abduction scoring (DAS) assay. A single administration of ouabain significantly prolonged BoNT/A-induced neuromuscular paralysis. Moreover, consecutive daily injection of ouabain exacerbated BoNT/A-induced neuromuscular paralysis, and led to a significant decrease in both twitch and tetanic forces as assayed in isolated BoNT/A-injected muscles. We next looked at the effects of ouabain on BoNT/A-induced muscle atrophy. Administration of ouabain led to a decrease in the myofibrillar cross-sectional area (CSAs) by 14 post-BoNT/A injection. In addition, repeated administration of ouabain increased mRNA expression levels of ubiquitin ligases, which are markers of muscle atrophy, in BoNT/A-injected muscle. These results suggest that ouabain exacerbates BoNT/A-induced neuromuscular paralysis via a marked progression of BoNT/A-induced muscle atrophy.
机译:肉毒杆菌神经毒素血清型A(BoNT / A)抑制离体肌肉中神经肌肉接头处的乙酰胆碱释放,哇巴因可以部分阻断其作用。然而,尚不清楚哇巴因是否在体内减弱BoNT / A引起的神经肌肉麻痹。在这项工作中,我们调查了哇巴因对BoNT / A诱导的小鼠神经肌肉麻痹的影响。将BoNT / A单次注入骨骼肌后,立即向小鼠腹膜内注射Ouabain。通过手指外展评分(DAS)分析定量监测肌肉张力和手指外展,评估哇巴因对BoNT / A诱导的肌肉麻痹的影响。一次单用哇巴因可显着延长BoNT / A引起的神经肌肉麻痹。此外,每天连续注射哇巴因会加剧BoNT / A诱导的神经肌肉麻痹,并导致抽搐和强直肌力的显着降低,如在分离的BoNT / A注射的肌肉中测定的那样。接下来,我们研究了哇巴因对BoNT / A引起的肌肉萎缩的影响。 BoNT / A后14次注射,哇巴因的使用导致肌原纤维横截面积(CSA)减少。另外,重复施用哇巴因在注射BoNT / A的肌肉中增加了泛素连接酶的mRNA表达水平,泛素连接酶是肌肉萎缩的标志。这些结果表明,哇巴因通过BoNT / A引起的肌肉萎缩的明显进展而加剧BoNT / A引起的神经肌肉麻痹。

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