Cigarette smoking induces skeletal muscle atrophy through TNF-α/Caspase-3/GSDME-mediated pyroptosis_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

TAN Yufen ¹ , XIONG Guolin ² , YE Yuanyuan ¹ , HUANG Cuibi ¹ , WEI Xinyan ¹ , LI Jie ¹ , HUANG Lihua ¹ ,  BIN Yanfei ¹

1. Department of Respiratory and Critical Care Medicine, The Second Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, 530007, P.R. China;
2. Department of Epidemiology, School of Public Health, Guangxi Medical University, Nanning, Guangxi, 530021, P.R. China;

Corresponding author：

BIN Yanfei, Email: binyanfei@163.com

Keywords：

Cigarette smoke; chronic obstructive pulmonary disease; skeletal muscle atrophy; cysteinyl aspartate specific proteinase-3; gasdermin-E; pyroptosis

DOI：

10.7507/1671-6205.202408075

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Abstract Full text Figures/Tables Video References Cited by

Objective To investigate the role of cysteinyl aspartate specific proteinase-3 (Caspase-3)/ gasdermin-E (GSDME)-mediated pyroptosis in skeletal muscle atrophy induced by cigarette smoke in mice.Methods To construct a mouse model of COPD, C57BL/6 mice were exposed to cigarette smoke (CS) for 24 weeks. HE staining was used to observe the changes in the morphology of the gastrocnemius muscle in mice. Immunohistochemistry was used to detect the expression of pyroptosis-related proteins in gastrocnemius muscle. To construct a model of skeletal muscle cell atrophy in vitro, C2C12 myoblasts were induced to differentiate into skeletal muscle cells with 2% horse serum, and then skeletal muscle cells were treated with cigarette smoke extract (CSE). Skeletal muscle cells were further treated with the caspase-3 inhibitor Z-DEVD-FMK and the GSDME inhibitor Dimethyl fumarate (DMF) to explore the effects of inhibition of caspase-3/GSDME on CSE-induced skeletal muscle cell atrophy. To observe the effects of TNF-α on the expression of caspase-3 and GSDME proteins as well as the impact on myotubes, skeletal muscle cells were stimulated with tumor necrosis factor-alpha (TNF-α). Western blotting was applied to detect protein expression levels of caspase-3 and GSDME in skeletal muscle cells. Hoechst 33342/ Hoechst33342/ Propidium Iodide (PI) staining was applied to detect the PI-positive rate of skeletal muscle cells. The lactate dehydrogenase (LDH) release of C2C12 myotubes was measured by LDH release test. Immunofluorescence was used to detect changes in myotube diameter. Results CS-induced skeletal muscle atrophy was observed in mice, accompanied by increased pyroptosis- associated proteins (c-caspase-3 and GSDME-N) (P<0.05). CSE also induced elevated c-caspase-3 and GSDME-N expression in C2C12 cells , resulting in increased LDH release, positive ratio of PI, along with reduced myotube diameter (P<0.05). In addition, TNF-α promotes myotube atrophy and the expression of cleaved-caspase-3 and GSDME-N proteins in skeletal muscle cells. Conclusion CS can induce skeletal muscle atrophy through activated TNF-α/Caspase-3/GSDME-mediated pyroptosis.

Citation： TAN Yufen, XIONG Guolin, YE Yuanyuan, HUANG Cuibi, WEI Xinyan, LI Jie, HUANG Lihua, BIN Yanfei. Cigarette smoking induces skeletal muscle atrophy through TNF-α/Caspase-3/GSDME-mediated pyroptosis. Chinese Journal of Respiratory and Critical Care Medicine, 2024, 23(10): 710-718. doi: 10.7507/1671-6205.202408075 Copy

1.	Xia S, Hollingsworth LR 4th, Wu H. Mechanism and Regulation of Gasdermin-Mediated Cell Death. Cold Spring Harb Perspect Biol, 2020, 12(3): a036400.
2.	Song Q, Chen P, Liu XM. The role of cigarette smoke-induced pulmonary vascular endothelial cell apoptosis in COPD. Respir Res, 2021, 22(1): 39.
3.	Sepúlveda-Loyola W, Osadnik C, Phu S, et al. Diagnosis, prevalence, and clinical impact of sarcopenia in COPD: a systematic review and meta-analysis. J Cachexia Sarcopenia Muscle, 2020, 11(5): 1164-1176.
4.	Swallow EB, Reyes D, Hopkinson NS, et al. Quadriceps strength predicts mortality in patients with moderate to severe chronic obstructive pulmonary disease. Thorax, 2007, 62(2): 115-120.
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6.	Liu D, Xiao Y, Zhou B, et al. PKM2-dependent glycolysis promotes skeletal muscle cell pyroptosis by activating the NLRP3 inflammasome in dermatomyositis/polymyositis. Rheumatology (Oxford), 2021, 60(5): 2177-2189.
7.	Aluganti Narasimhulu C, Singla DK. Amelioration of diabetes-induced inflammation mediated pyroptosis, sarcopenia, and adverse muscle remodelling by bone morphogenetic protein-7. J Cachexia Sarcopenia Muscle, 2021, 12(2): 403-420.
8.	Wang L, Jiao XF, Wu C, et al. Trimetazidine attenuates dexamethasone-induced muscle atrophy via inhibiting NLRP3/GSDMD pathway-mediated pyroptosis. Cell Death Discov, 2021, 7(1): 251.
9.	Li YQ, Peng JJ, Peng J, et al. The deafness gene GSDME: its involvement in cell apoptosis, secondary necrosis, and cancers. Naunyn Schmiedebergs Arch Pharmacol, 2019, 392(9): 1043-1048.
10.	Wang Y, Gao W, Shi X, et al. Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin. Nature, 2017, 547(7661): 99-103.
11.	Wu J, Lin S, Chen W, et al. TNF-α contributes to sarcopenia through caspase-8/caspase-3/GSDME-mediated pyroptosis. Cell Death Discov, 2023, 9(1): 76.
12.	Fan X, Dong T, Yan K, et al. PM2. 5 increases susceptibility to acute exacerbation of COPD via NOX4/Nrf2 redox imbalance-mediated mitophagy. Redox Biol, 2022, 59: 102587.
13.	刘文婷, 苏文燕, 杨霞, 等. 香烟烟雾提取物通过氧化应激下调组蛋白去乙酰化酶2诱导小鼠骨骼肌细胞早衰. 细胞与分子免疫学杂志, 2015, 31(5): 630-633.
14.	Ono Y, Maejima Y, Saito M, et al. TAK-242, a specific inhibitor of Toll-like receptor 4 signalling, prevents endotoxemia-induced skeletal muscle wasting in mice. Sci Rep, 2020, 10: 694.
15.	Jaitovich A, Barreiro E. Skeletal Muscle Dysfunction in Chronic Obstructive Pulmonary Disease. What We Know and Can Do for Our Patients. Am J Respir Crit Care Med, 2018, 198(2): 175-186.
16.	Degens H, Gayan-Ramirez G, van Hees HW. Smoking-induced skeletal muscle dysfunction: from evidence to mechanisms. Am J Respir Crit Care Med, 2015, 191(6): 620-625.
17.	Bin Y, Xiao Y, Huang D, et al. Theophylline inhibits cigarette smoke-induced inflammation in skeletal muscle by upregulating HDAC2 expression and decreasing NF-κB activation. Am J Physiol Lung Cell Mol Physiol, 2019, 316(1): L197-L205.
18.	Galluzzi L, Vitale I, Aaronson SA, et al. Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ, 2018, 25(3): 486-541.
19.	Li W, Sun J, Zhou X, et al. Mini-Review: GSDME-Mediated Pyroptosis in Diabetic Nephropathy. Front Pharmacol, 2021, 12: 780790.
20.	Jiang M, Qi L, Li L, et al. The caspase-3/GSDME signal pathway as a switch between apoptosis and pyroptosis in cancer. Cell Death Discov, 2020, 6: 112.
21.	Ding J, Li F, Cong Y, et al. Trichostatin A inhibits skeletal muscle atrophy induced by cigarette smoke exposure in mice. Life Sci, 2019, 235: 116800.
22.	Frank D, Vince JE. Pyroptosis versus necroptosis: similarities, differences, and crosstalk. Cell Death Differ, 2019, 26(1): 99-114.
23.	McGeough MD, Wree A, Inzaugarat ME, et al. TNF regulates transcription of NLRP3 inflammasome components and inflammatory molecules in cryopyrinopathies. J Clin Invest, 2017, 127(12): 4488-4497.

1. Xia S, Hollingsworth LR 4th, Wu H. Mechanism and Regulation of Gasdermin-Mediated Cell Death. Cold Spring Harb Perspect Biol, 2020, 12(3): a036400.
2. Song Q, Chen P, Liu XM. The role of cigarette smoke-induced pulmonary vascular endothelial cell apoptosis in COPD. Respir Res, 2021, 22(1): 39.
3. Sepúlveda-Loyola W, Osadnik C, Phu S, et al. Diagnosis, prevalence, and clinical impact of sarcopenia in COPD: a systematic review and meta-analysis. J Cachexia Sarcopenia Muscle, 2020, 11(5): 1164-1176.
4. Swallow EB, Reyes D, Hopkinson NS, et al. Quadriceps strength predicts mortality in patients with moderate to severe chronic obstructive pulmonary disease. Thorax, 2007, 62(2): 115-120.
5. Shrikrishna D, Patel M, Tanner R J, et al. Quadriceps wasting and physical inactivity in patients with COPD. Eur Respir J, 2012, 40(5): 1115-1122.
6. Liu D, Xiao Y, Zhou B, et al. PKM2-dependent glycolysis promotes skeletal muscle cell pyroptosis by activating the NLRP3 inflammasome in dermatomyositis/polymyositis. Rheumatology (Oxford), 2021, 60(5): 2177-2189.
7. Aluganti Narasimhulu C, Singla DK. Amelioration of diabetes-induced inflammation mediated pyroptosis, sarcopenia, and adverse muscle remodelling by bone morphogenetic protein-7. J Cachexia Sarcopenia Muscle, 2021, 12(2): 403-420.
8. Wang L, Jiao XF, Wu C, et al. Trimetazidine attenuates dexamethasone-induced muscle atrophy via inhibiting NLRP3/GSDMD pathway-mediated pyroptosis. Cell Death Discov, 2021, 7(1): 251.
9. Li YQ, Peng JJ, Peng J, et al. The deafness gene GSDME: its involvement in cell apoptosis, secondary necrosis, and cancers. Naunyn Schmiedebergs Arch Pharmacol, 2019, 392(9): 1043-1048.
10. Wang Y, Gao W, Shi X, et al. Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin. Nature, 2017, 547(7661): 99-103.
11. Wu J, Lin S, Chen W, et al. TNF-α contributes to sarcopenia through caspase-8/caspase-3/GSDME-mediated pyroptosis. Cell Death Discov, 2023, 9(1): 76.
12. Fan X, Dong T, Yan K, et al. PM2. 5 increases susceptibility to acute exacerbation of COPD via NOX4/Nrf2 redox imbalance-mediated mitophagy. Redox Biol, 2022, 59: 102587.
13. 刘文婷, 苏文燕, 杨霞, 等. 香烟烟雾提取物通过氧化应激下调组蛋白去乙酰化酶2诱导小鼠骨骼肌细胞早衰. 细胞与分子免疫学杂志, 2015, 31(5): 630-633.
14. Ono Y, Maejima Y, Saito M, et al. TAK-242, a specific inhibitor of Toll-like receptor 4 signalling, prevents endotoxemia-induced skeletal muscle wasting in mice. Sci Rep, 2020, 10: 694.
15. Jaitovich A, Barreiro E. Skeletal Muscle Dysfunction in Chronic Obstructive Pulmonary Disease. What We Know and Can Do for Our Patients. Am J Respir Crit Care Med, 2018, 198(2): 175-186.
16. Degens H, Gayan-Ramirez G, van Hees HW. Smoking-induced skeletal muscle dysfunction: from evidence to mechanisms. Am J Respir Crit Care Med, 2015, 191(6): 620-625.
17. Bin Y, Xiao Y, Huang D, et al. Theophylline inhibits cigarette smoke-induced inflammation in skeletal muscle by upregulating HDAC2 expression and decreasing NF-κB activation. Am J Physiol Lung Cell Mol Physiol, 2019, 316(1): L197-L205.
18. Galluzzi L, Vitale I, Aaronson SA, et al. Molecular mechanisms of cell death: recommendations of the Nomenclature Committee on Cell Death 2018. Cell Death Differ, 2018, 25(3): 486-541.
19. Li W, Sun J, Zhou X, et al. Mini-Review: GSDME-Mediated Pyroptosis in Diabetic Nephropathy. Front Pharmacol, 2021, 12: 780790.
20. Jiang M, Qi L, Li L, et al. The caspase-3/GSDME signal pathway as a switch between apoptosis and pyroptosis in cancer. Cell Death Discov, 2020, 6: 112.
21. Ding J, Li F, Cong Y, et al. Trichostatin A inhibits skeletal muscle atrophy induced by cigarette smoke exposure in mice. Life Sci, 2019, 235: 116800.
22. Frank D, Vince JE. Pyroptosis versus necroptosis: similarities, differences, and crosstalk. Cell Death Differ, 2019, 26(1): 99-114.
23. McGeough MD, Wree A, Inzaugarat ME, et al. TNF regulates transcription of NLRP3 inflammasome components and inflammatory molecules in cryopyrinopathies. J Clin Invest, 2017, 127(12): 4488-4497.

Chinese Journal of Respiratory and Critical Care Medicine

Cigarette smoking induces skeletal muscle atrophy through TNF-α/Caspase-3/GSDME-mediated pyroptosis

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