Effect of GSDMD-mediated pyroptosis on muscle atrophy of skeletal muscle in mice and its potential mechanism

Mei LI; Shuang LI; Jian HU; Zhengxiao LIN; Guirong ZHANG; Sha LIU; Fuxiang LI

doi:10.16016/j.1000-5404.202101229

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Effect of GSDMD-mediated pyroptosis on muscle atrophy of skeletal muscle in mice and its potential mechanism

Basic Medicine | 更新时间：2026-01-09

- Effect of GSDMD-mediated pyroptosis on muscle atrophy of skeletal muscle in mice and its potential mechanism
- Journal of Third Military Medical University Vol. 43, Issue 15, Pages: 1486-1492(2021)
- 作者机构：
  
  1.610031 成都, 西南交通大学医学院
  2.610083 成都, 中国人民解放军西部战区总医院: 重症医学科
  3.610083 成都, 中国人民解放军西部战区总医院: 心内科
- 作者简介：
- 基金信息：
  
  the National Nature Science Foundation for Young Scholars of China(81800338);the Military Medical Research Project of General Hospital of Western Theater Command(2019LH05)
- DOI：10.16016/j.1000-5404.202101229
  CLC： R329.27; R363.21; R746.4
- Received：27 January 2021，
  
  Revised：2021-04-30，
  
  Published：15 August 2021
- 稿件说明：
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Mei LI, Shuang LI, Jian HU, et al. Effect of GSDMD-mediated pyroptosis on muscle atrophy of skeletal muscle in mice and its potential mechanism[J]. Journal of Third Military Medical University, 2021, 43(15): 1486-1492.
DOI：

Mei LI, Shuang LI, Jian HU, et al. Effect of GSDMD-mediated pyroptosis on muscle atrophy of skeletal muscle in mice and its potential mechanism[J]. Journal of Third Military Medical University, 2021, 43(15): 1486-1492. DOI： 10.16016/j.1000-5404.202101229.

摘要

目的

探讨脂多糖(lipopolysaccharide，LPS)刺激下小鼠骨骼肌肌管中肌肉萎缩和GSDMD介导的细胞焦亡发生情况，以期为进一步阐明ICU获得性衰弱(ICU-acquired weakness

ICU-AW)的发病机制提供线索。

方法

将C2C12小鼠骨骼肌成肌细胞诱导分化形成肌管。分别设置: Control组，加无菌PBS溶液；LPS组，LPS处理浓度梯度为1、10、100 μg/mL；LPS+ATP组，以LPS组相同浓度梯度干预肌管后均添加ATP(5 mmol/L)。并分别在LPS干预6 h和12 h这两个时间点检测肌管内外发生的变化。光镜下观察肌管形态学变化，采用Western blot法检测各组肌管中肌管形成标志蛋白MHC、Myogenin、HDAC1和细胞焦亡标志蛋白GSDMD的表达，采用荧光定量PCR检测各组肌管中萎缩基因Atrogin-1、MuRF-1 mRNA和细胞焦亡相关炎性因子IL-18 mRNA表达，采用ELISA检测肌管培养上清IL-18浓度。

结果

① 光镜可见分化第5天肌管形成。②RT-qPCR检测结果显示: LPS干预12 h后LPS/LPS+ATP各组肌管中萎缩基因Atrogin-1和MuRF-1 mRNA的表达均显著上调(

0.05)，与细胞焦亡相关的炎性因子IL-18 mRNA在肌管中的表达趋势相同(

0.05)。③ELISA检测结果显示: LPS干预12h后各组肌管培养液上清中IL-18浓度显著高于对照组(

0.05)。④Western blot法检测结果显示: 各组干预的肌管中肌管形成标志蛋白MHC、Myogenin、HDAC1均能稳定表达；LPS/LPS+ATP处理后肌管GSDMD蛋白表达量较对照组显著增加(

0.05)。

结论

在LPS刺激下小鼠骨骼肌肌管中可发生细胞焦亡，并与肌管萎缩相关。

Abstract

Objective

To investigate the occurrence of gasdermin D protein (GSDMD)-mediated pyroptosis and muscle atrophy in mice under lipopolysaccharide (LPS) stimulation in order to elucidate the possible pathogenetic mechanisms of intensive care unit-acquired weakness (ICU-AW).

Methods

The mouse skeletal muscle myoblasts (C2C12) were induced to differentiate into myotubes

and then divided into groups as follows: control group (sterile PBS solution)

LPS groups (1

10 and 100 μg/mL)

and LPS+ATP groups (5 mmol/L ATP was added to the corresponding LPS gradient groups). The changes inside and outside the myotubes were detected and compared among groups at the time points of 6 or 12 h after LPS intervention. The morphological changes of myotubes were observed under light microscope; Western blotting was used to determine the myotube formation marker proteins MHC

myogenin and HDAC1

as well as pyroptosis-related protein

GSDMD

in each group; RT-qPCR was employed to test the mRNA expression of atrophic genes Atrogin-1

MuRF-1 and pyroptosis-related proinflammatory cytokine IL-18 in the myotubes of each group; and the IL-18 concentration in the supernatant of myotube culture was detected by ELISA.

Results

Myotubes formation was observed on day 5 of differentiation. Under the stimulation of LPS/LPS+ATP

the mRNA levels of Atrogin-1 and MuRF-1 were significantly up-regulated (

0.05)

and the expression of IL-18 presented a same trend (

0.05). ELISA results showed that after LPS intervention for 12 h

the concentration of IL-18 in the myotube culture supernatant was significantly higher in the intervention groups than the control group (

0.05). In addition

the levels of MHC

Myogenin and HDAC1 proteins were stably expressed in the intervention groups

and the expression of GSDMD protein in the LPS/LPS+ATP groups were significantly increased compared with the control group (

0.05).

Conclusion

LPS induces cell pyroptosis in the myotubes of mouse skeletal muscle

which is correlated with the occurrence of muscle atrophy.

关键词

Keywords

references

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