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1.遵义医科大学人体解剖学教研室,贵州 遵义 563000
2.遵义医科大学2022级第一临床学院,贵州 遵义 563000
王猛,E-mail:932051454@qq.com
收稿:2025-02-05,
纸质出版:2026-02-25
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王猛,李旭灿,王宇等.甘氨酰-tRNA合成酶肌内注射改善小鼠失神经肌萎缩的机制研究[J].局解手术学杂志,2026,35(02):127-132.
WANG Meng,LI Xu-can,WANG Yu,et al.Mechanism of intramuscular injection of glycyl-tRNA synthetase in improving denervated muscular atrophy in mice[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2026,35(02):127-132.
王猛,李旭灿,王宇等.甘氨酰-tRNA合成酶肌内注射改善小鼠失神经肌萎缩的机制研究[J].局解手术学杂志,2026,35(02):127-132. DOI: 10.11659/jjssx.02E025007.
WANG Meng,LI Xu-can,WANG Yu,et al.Mechanism of intramuscular injection of glycyl-tRNA synthetase in improving denervated muscular atrophy in mice[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2026,35(02):127-132. DOI: 10.11659/jjssx.02E025007.
目的
2
探讨甘氨酰-tRNA合成酶(GlyRS)肌内注射改善小鼠失神经肌萎缩的作用机制。
方法
2
构建GlyRS过表达腺病毒载体,进行PCR鉴定和测序。将54只雄性SPF级昆明小鼠随机分为对照(C)组(暴露坐骨神经,但不离断损伤)、坐骨神经离断损伤(SNI)组(构建SNI致胫骨前肌萎缩模型)、SNI+NC组(建模后转染空病毒载体)、SNI+GlyRS组(建模后转染GlyRS过表达腺病毒载体),每组12只,其余6只小鼠胫骨前肌作冰冻切片。荧光显微镜检测GlyRS过表达腺病毒载体转染昆明小鼠胫骨前肌的效果;测量各组小鼠胫骨前肌肌重和肌纤维横截面积。qRT-PCR和Western blot检测各组小鼠胫骨前肌的GlyRS、mTOR的mRNA和蛋白表达水平。
结果
2
GlyRS过表达腺病毒载体经PCR鉴定和测序正确,并成功转染小鼠患侧胫骨前肌。与C组相比,SNI组小鼠胫骨前肌肌重和肌纤维横截面积明显减少(
P
<
0.01),GlyRS和mTOR的mRNA和蛋白表达水平升高(
P
<
0.01);与SNI组相比,SNI+NC组小鼠胫骨前肌肌重和肌纤维横截面积无显著变化(
P
>
0.05),GlyRS和mTOR的mRNA和蛋白表达水平升高(
P
<
0.01),SNI+GlyRS组小鼠胫骨前肌肌重和肌纤维横截面积明显增加(
P
<
0.01),GlyRS和mTOR的mRNA和蛋白表达水平明显升高(
P
<
0.01);与SNI+NC组相比,SNI+GlyRS组小鼠胫骨前肌肌重和肌纤维横截面积明显增加(
P
<
0.01),GlyRS和mTOR的mRNA和蛋白表达水平明显升高(
P
<
0.01)。
结论
2
GlyRS肌内注射可能通过促进mTOR的合成改善小鼠坐骨神经离断损伤所致的胫骨前肌萎缩。
Objective
2
To investigate the mechanism of intramuscular injection of glycyl tRNA synthetase (GlyRS) in improving denervated muscular atrophy in mice.
Methods
2
GlyRS overexpression adenovirus vector was constructed for PCR identification and sequencing. A total of 54 male SPF Kunming mice were randomly divided into control (C) group (the sciatic nerve was exposed, but not transection injury), sciatic nerve transection injury(SNI) group (constructing a model of tibialis anterior muscle atrophy induced by SNI), SNI+NC group (transfected an empty viral vector after modeling), and SNI+GlyRS group (transfected with GlyRS overexpression adenoviral vector after modeling), with 12 mice in each group; and the tibialis anterior muscle of the remaining 6 mice were used to make frozen sections. Fluorescence microscope was used to detect the transfection effect of GlyRS overexpression adenovirus vector on tibialis anterior muscle of Kunming mice. Tibialis anterior muscle weight and muscle fiber cross-sectional area of mice in each group were measured. qRT-PCR and Western blot were used to detect the mRNA and protein expression levels of GlyRS and mTOR in tibialis anterior muscle of mice in each group.
Results
2
The GlyRS overexpression adenoviral vector was identified and sequenced correctly by PCR,and successfully transfected into the affected tibialis anterior muscle of mice. Compared with group C, the tibialis anterior muscle weight and muscle fiber cross-sectional area of mice
in SNI group were significantly decreased (
P
<
0.01), and the mRNA and protein expression levels of GlyRS and mTOR were increased (
P
<
0.01). Compared with SNI group, the tibialis anterior muscle weight and muscle fiber cross-sectional area of mice in SNI+NC group had no significant changes (
P
>
0.05), and the mRNA and protein expression levels of GlyRS and mTOR were increased (
P
<
0.01);the tibialis anterior muscle weight and muscle fiber cross-sectional area of mice in the SNI+GlyRS group were significantly increased (
P
<
0.01), and the mRNA and protein expression levels of GlyRS and mTOR were significantly increased (
P
<
0.01). Compared with SNI+NC group, the tibialis anterior muscle weight and muscle fiber cross-sectional area of mice in SNI+GlyRS group were significantly increased (
P
<
0.01), and the mRNA and protein expression levels of GlyRS and mTOR were significantly increased (
P
<
0.01).
Conclusion
2
Intramuscular injection of GlyRS may improve tibialis anterior muscle atrophy induced by sciatic nerve transection injury in mice by promoting the synthesis of mTOR.
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