目的 探讨推拿延缓失神经骨骼肌萎缩的相关机制。方法 77只Sprague-Dawley雄性大鼠随机分为假手术组(n = 7)、模型组(n = 35)和推拿组(n = 35)。后两组通过暴露并切断胫神经的方法制备失神经骨骼肌萎缩大鼠模型。造模后第2天，推拿组术侧下肢进行推拿干预。分别在造模后0 d、7 d、14 d、21 d和28 d取材，测定术侧腓肠肌湿重比，HE染色测定腓肠肌肌纤维截面积和直径，逆转录实时定量聚合酶链反应检测术侧腓肠肌组织中自噬相关因子Beclin-1、液泡分选蛋白34 (Vps34)和微管相关蛋白轻链3 (LC3)的mRNA表达。结果 三组0 d时，腓肠肌湿重比、肌纤维截面积和直径，以及Beclin-1、Vps34和LC3的mRNA表达均无显著性差异(F < 1.321, P > 0.05)。与假手术组比较，模型组和推拿组腓肠肌湿重比、肌纤维截面积和直径在不同时间下降(P < 0.05)，推拿组肌湿重比(除21 d)均高于模型组(P < 0.05)，推拿组肌纤维截面积和直径(除21 d)均大于模型组(P < 0.05)。与假手术组相比，模型组和推拿组术侧腓肠肌Beclin-1、Vps34、LC3 mRNA表达在不同时间升高(P < 0.05)，推拿组各项mRNA表达(除14 d)均高于模型组(P < 0.05)。组内比较，模型组和推拿组腓肠肌湿重比、肌纤维截面积和直径随时间呈进行性下降趋势(P < 0.05)；模型组Beclin-1、Vps34 mRNA表达在21 d后呈上升趋势(P < 0.05)，LC3 mRNA表达在21 d升高(P < 0.05)；推拿组Beclin-1 mRNA表达呈先上升后下降的趋势(P < 0.05)，Vps34和LC3 mRNA表达(除14 d)呈先上升后下降趋势(P < 0.05)。结论 推拿可能通过上调自噬相关因子Beclin-1、Vps34和LC3的mRNA表达，促进细胞自噬的激活，清除损伤细胞器和蛋白质，为肌纤维再生提供一定的合成底物和能量，从而减轻失神经骨骼肌萎缩的程度。
安荟羽
,
唐成林
,
黄思琴
,
赵丹丹
,
罗翱
,
吴梦佳
,
谭程方
,
邱丽
,
万小凤
,
马翔
. 推拿对失神经骨骼肌萎缩大鼠自噬相关因子Beclin-1、液泡分选蛋白34和微管相关蛋白轻链3的mRNA表达的影响[J]. 中国康复理论与实践, 2019
, 25(2)
: 184
-191
.
DOI: 10.3969/j.issn.1006-9771.2019.02.010
Objective To explore the effect and mechanism of Tuina on denervated skeletal muscle atrophy.
Methods A total of 77 male Sprague-Dawley rats were randomly divided into sham group (n = 7), model group (n = 35) and Tuina group (n = 35). The latter two groups were established skeletal muscle atrophy model by exposing and cutting off the common tibial nerve of rats. One day after modeling, the lower limbs of the surgical side received Tuina in Tuina group. Separately, the surgical side of gastrocnemius muscle were sampled on the 0th, 7th, 14th, 21st and 28th day after modeling, and measured the wet mass ratio. The cross-sectional area and diameter of muscle fiber were measured after HE staining. The mRNA expression of autophagy-realated factor Beclin-1, vacuolar protein sorting (Vps34) and microtubule-associated protein light chain 3 (LC3) were tested with reverse transcription real-time quantitative polymerase chain reaction.
Results There was no statistical difference in the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber, and the mRNA expression of Beclin-1, Vps34 and LC3 among three groups on the 0th day (F < 1.321, P > 0.05). Compared with the sham group, the ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber decreased at different time points in the model group and Tuina group (P < 0.05), the ratio of gastrocnemius wet weight was higher, and the cross-sectional area and diameter of muscle fiber were bigger, both except on the 21st day, in Tuina group than in the model group (P < 0.05). Compared with the sham group, the mRNA expression of Beclin-1, Vps34 and LC3 increased at different points in the model group than in Tuina group (P < 0.05), and all the mRNA expression was higher, except on the 14th day, in Tuina group than in the model group (P < 0.05). The ratio of gastrocnemius wet weight, the cross-sectional area and diameter of muscle fiber showed a trend of progressive decrease with time in the model group and Tuina group (P < 0.05). The mRNA expression of Beclin-1 and Vps34 increased (P < 0.05), and the mRNA expression of LC3 increased in the model group 21 days after intervention (P < 0.05). The mRNA expression of Beclin-1, Vps34 and LC3 increased first and then decreased, except the mRNA expression on the 14th day in Tuina group (P < 0.05).
Conclusion Tuina may promote the activation of autophagy by up-regulating the expression of autophagy-realated factor Beclin-1, Vps34 and LC3, remove the damaged organelles and proteins, provide certain synthetic substrate and energy for muscle fiber regeneration, thereby reduce the loss of degree of denervated skeletal muscle atrophy.
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