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[电针通过调节坐骨神经损伤大鼠中叉头蛋白3A、肌肉萎缩F盒蛋白和生肌分化抗原蛋白的表达改善失神经支配的腓肠肌萎缩]

[Electroacupuncture improves denervated gastrocnemius atrophy by regulating expression of fork-head protein 3A,muscle atrophy F-box and myogenic differentiation antigen proteins in sciatic nerve injury rats].

作者信息

Wu Meng-Jia, Tang Cheng-Lin, Huang Si-Qin, Zhao Dan-Dan, Luo Ao, Zhang An-Ning, An Hui-Yu, Tan Cheng-Fang, Qiu Li

机构信息

College of Traditional Chinese Medicine, Chongqing Medical University, Chongqing 400016, China; Chongqing Key Laboratory of Traditional Chinese Medicine for Prevention and Treatment of Metabolic Diseases, Chongqing Medical University, Chongqing 400016, China.

出版信息

Zhen Ci Yan Jiu. 2019 Apr 25;44(4):253-7. doi: 10.13702/j.1000-0607.171015.

Abstract

OBJECTIVE

To observe the effect of electroacupuncture (EA) on morphological changes of denervated gastrocnemius(GS) and the expression of fork-head protein(FOXO3A), muscle atrophy F-box(MAFbx)and myogenic differentiation antigen (Myod1) in sciatic nerve injury rats, so as to reveal its mechanism underlying improvement of myoatrophy.

METHODS

Eighteen male Sprague-Dawley rats were randomly divided into sham operation, model and EA groups (=6 per group). The model of gastrocnemius atrophy was established by crushing the right sciatic nerve. Then, EA (2 Hz) was applied to the right "Zusanli" (ST36) and "Huantiao" (GB30) for 10 min, once a day for 14 successive days. The wet weight of the GS on both sides was weighted to calculate the wet weight ratio (the injured side /the healthy side), and the cross-sectional area (CSA) and diameter of GS fibers were measured after H.E. staining. The expressions of FOXO3A, MAFbx and Myod1 protein and mRNA in the GS tissue were tested using Western blot and fluorescence quantitative PCR, separately.

RESULTS

Following modeling, the GS wet weight ratio, CSA and fiber diameter were smaller in the model group than those in the sham group (<0.01), and were significantly higher in the EA group than in the model group (<0.01). H.E. staining showed that the GS fibers became smaller and the myocyte got round in the model group, while the GS fibers were bigger and the myocyte was relatively regular in morphology in the EA group. After modeling, the expression levels of FOXO3A, MAFbx and Myod1 mRNA and protein were evidently higher in the model group (<0.01); Moreover, after EA treatment, modeling-induced increasing of expression levels of FOXO3A and MAFbx mRNA and protein were revised (<0.01), while the increased expression level of Myod1 was further up-regulated relavant to that in the model group (<0.01)..

CONCLUSION

EA of ST36 and GB30 can suppress the up-regulated expression of FOXO3A and MAFbx mRNA and protein and further promote the expression of Myod1 mRNA and protein in the GS tissue in rats with denervated GS atrophy, which may contribute to its function in relieving the myoatrophy, promoting the skeletal muscle protein hydrolysis and differentiation of satellite cells.

摘要

目的

观察电针(EA)对坐骨神经损伤大鼠失神经支配腓肠肌(GS)形态学变化及叉头蛋白(FOXO3A)、肌肉萎缩F盒蛋白(MAFbx)和生肌分化抗原(Myod1)表达的影响,以揭示其改善肌萎缩的机制。

方法

将18只雄性Sprague-Dawley大鼠随机分为假手术组、模型组和电针组(每组6只)。通过钳夹右侧坐骨神经建立腓肠肌萎缩模型。然后,将电针(2Hz)施加于右侧“足三里”(ST36)和“环跳”(GB30)10分钟,每天1次,连续14天。称取双侧GS的湿重以计算湿重比(损伤侧/健侧),苏木精-伊红(H.E.)染色后测量GS纤维的横截面积(CSA)和直径。分别采用蛋白质免疫印迹法和荧光定量PCR检测GS组织中FOXO3A、MAFbx和Myod1蛋白及mRNA的表达。

结果

建模后,模型组GS的湿重比、CSA和纤维直径均小于假手术组(P<0.01),电针组显著高于模型组(P<0.01)。H.E.染色显示,模型组GS纤维变细,肌细胞呈圆形,而电针组GS纤维较粗大,肌细胞形态相对规则。建模后,模型组FOXO3A、MAFbx和Myod1 mRNA及蛋白表达水平明显升高(P<0.01);此外,电针治疗后,建模诱导的FOXO3A和MAFbx mRNA及蛋白表达水平升高得到纠正(P<0.01),而Myod1表达水平的升高较模型组进一步上调(P<0.01)。

结论

电针ST36和GB30可抑制失神经支配GS萎缩大鼠GS组织中FOXO3A和MAFbx mRNA及蛋白的上调表达,并进一步促进Myod1 mRNA及蛋白的表达,这可能有助于其减轻肌萎缩、促进骨骼肌蛋白水解及卫星细胞分化的作用。

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