Gigliotti Deanna, Leiter Jeff R S, Macek Bryce, Davidson Michael J, MacDonald Peter B, Anderson Judy E
Department of Biological Sciences, University of Manitoba, Winnipeg, Manitoba, Canada;
Pan Am Clinic, Winnipeg, Manitoba, Canada.
Am J Physiol Cell Physiol. 2015 Sep 15;309(6):C383-91. doi: 10.1152/ajpcell.00143.2015. Epub 2015 Jul 1.
The high frequency of poor outcome and chronic pain after surgical repair of shoulder rotator-cuff injury (RCI) prompted this study to explore the potential to amplify muscle regeneration using nitric oxide (NO)-based treatment. After preoperative magnetic resonance imaging (MRI), biopsies of supraspinatus and ipsilateral deltoid (as a control) were collected during reparative surgery for RCI. Muscle fiber diameter, the pattern of neuromuscular junctions observed with alpha-bungarotoxin staining, and the γ:ε subunit ratio of acetylcholine receptors in Western blots were examined in tandem with experiments to determine the in vitro responsiveness of muscle satellite cells to activation (indicated by uptake of bromodeoxyuridine, BrdU) by the NO-donor drug, isosorbide dinitrate (ISDN). Consistent with MRI findings of supraspinatus atrophy (reduced occupation ratio and tangent sign), fiber diameter was lower in supraspinatus than in deltoid. ISDN induced a significant increase over baseline (up to 1.8-fold), in the proportion of BrdU+ (activated) Pax7+ satellite cells in supraspinatus, but not in deltoid, after 40 h in culture. The novel application of denervation indices revealed a trend for supraspinatus muscle to have a higher γ:ε subunit ratio than deltoid (P = 0.13); this ratio inversely with both occupancy ratio (P < 0.05) and the proportion of clusters at neuromuscular junctions (P = 0.05). Results implicate possible supraspinatus denervation in RCI and suggest NO-donor treatment has potential to promote growth in atrophic supraspinatus muscle after RCI and improve functional outcome.
肩袖损伤(RCI)手术修复后不良预后和慢性疼痛的高发生率促使本研究探索使用基于一氧化氮(NO)的治疗方法来增强肌肉再生的潜力。术前进行磁共振成像(MRI)后,在RCI修复手术期间采集冈上肌和同侧三角肌(作为对照)的活检样本。在进行实验以确定肌肉卫星细胞对NO供体药物二硝酸异山梨酯(ISDN)激活的体外反应性(以溴脱氧尿苷BrdU摄取表示)的同时,检测肌纤维直径、用α-银环蛇毒素染色观察到的神经肌肉接头模式以及Western印迹中乙酰胆碱受体的γ:ε亚基比率。与冈上肌萎缩的MRI结果(占有率降低和切线征)一致,冈上肌的纤维直径低于三角肌。培养40小时后,ISDN使冈上肌中BrdU +(激活的)Pax7 +卫星细胞的比例相对于基线显著增加(高达1.8倍),而三角肌中则没有。去神经指数的新应用显示冈上肌的γ:ε亚基比率有高于三角肌的趋势(P = 0.13);该比率与占有率(P < 0.05)和神经肌肉接头处簇的比例(P = 0.05)均呈负相关。结果表明RCI中可能存在冈上肌去神经支配,并提示NO供体治疗有可能促进RCI后萎缩的冈上肌生长并改善功能预后。