Regulation effect of endoplasmic reticulum stress and unfolded protein response activation on skeletal muscle fiber immune behavior

HUANG Jing-Wen; ZHANG Ren-Fei; JIAN Xiao-Ting; LIAO Zhao-Hong; LIAO Zhao-Hong Hai-Qiang; HUANG Tao; HU Ji-Jie; LIAO Hua

doi:10.16098/j.issn.0529-1356.2022.06.006

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Acta Anatomica Sinica ›› 2022, Vol. 53 ›› Issue (6) : 727-736. DOI: 10.16098/j.issn.0529-1356.2022.06.006

Regulation effect of endoplasmic reticulum stress and unfolded protein response activation on skeletal muscle fiber immune behavior

HUANG Jing-wen¹ ZHANG Ren-fei² JIAN Xiao-ting¹ LIAO Zhao-hong¹ LAN Hai-qiang¹ HUANG Tao¹ HU Ji-jie^3* LIAO Hua^1*#br#

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Abstract

Objective To investigate the regulatory effects of activated endoplasmic reticulum stress (ERS) and unfolded protein response (UPR) on the immune behavior of the stressed muscle fibers in inflammatory environments induced by interferon-γ(IFN-γ). Methods The myogenic precursor cells (MPCs) of C57 BL/6 mice cultured in vitro were differentiated into multinucleated myogenic tubes by horse serum and then to set up: 1. Control group; 2. IFN-γ group; 3. Tunicamycin group; 4. Thapsigargin group; 5. IFN-γ and 4-phenylbutyrate(4-PBA) combined treatment group; 6. IFN-γ, TG and 4-PBA combined treatment group; 7. IFN-γ and 4μ8c combined treatment group; 8. IFN-γ, TG and 4μ8c combined treatment group; 9. IFN-γ and GSK2606414 combined treatment group; 10. IFN-γ, TG and GSK2606414 combined treatment group. The level of myokines gene was detected by Real-time PCR. The expression of UPR key molecules including eukaryotic intiatio factor 2α(eIF2α), inositol requrring enzyme 1α(IRE1α) and activating transcription factor 6(ATF6) in muscle fibers was observed by immunofluorescence. Western blotting was used to detect immune molecules related to muscle cells, myokines and key molecules of UPR. Luminex analyzed the levels of pro-inflammatory myokines in muscle fibers. Results The expression of H-2Kb, H2-Ea, Toll like receptor 3(TLR3), p-eIF2α and p-IRE1α were up-regulated in IFN-γ induced inflammatory environment. The expression of H-2Kb, H2-Ea, TLR3 and myokines in the group with UPR inhibitor 4-PBA was down-regulated compared with IFN-γ group, and the expression of these molecules in the group with IRE1α specific inhibitor 4μ8c was down-regulated compared with the IFN-γ group. The addition of protein kinase R-like endoplasmic eticulum(PERK) specific inhibitor GSK2606414 showed no significant change. Conclusion In IFN-γ induced inflammatory environment, the UPR-IRE1α pathway activates and inhibits the synthesis of muscle fiber immune-related molecules, which further inhibits the muscle fiber mediated immune response and facilitates muscle regeneration.

Key words

Muscle fiber / Endoplasmic reticulum stress / Unfolded protein response / Inositol requiring enzyme 1α / Primary cell culture / Western blotting / Mouse

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HUANG Jing-wen ZHANG Ren-fei JIAN Xiao-ting LIAO Zhao-hong LAN Hai-qiang HUANG Tao HU Ji-jie LIAO Hua.

Regulation effect of endoplasmic reticulum stress and unfolded protein response activation on skeletal muscle fiber immune behavior

[J]. Acta Anatomica Sinica. 2022, 53(6): 727-736 https://doi.org/10.16098/j.issn.0529-1356.2022.06.006

References

［1］Kim I, Xu W, Reed JC. Cell death and endoplasmic reticulum stress: disease relevance and therapeutic opportunities［J］. Nat Rev Drug Discov, 2008, 7(12): 1013-1030.

［2］Bettigole SE, Glimcher LH. Endoplasmic reticulum stress in immunity［J］. Annu Rev Immunol, 2015, 33: 107-138.

［3］Rayavarapu S, Coley W, Nagaraju K. Endoplasmic reticulum stress in skeletal muscle homeostasis and disease［J］. Curr Rheumatol Rep, 2012, 14(3): 238-243.

［4］Vitadello M, Doria A, Tarricone E, et al. Myofiber stress-response in myositis: parallel investigations on patients and experimental animal models of muscle regeneration and systemic inflammation［J］. Arthritis Res Ther, 2010, 12(2): R52.

［5］Sciorati C, Monno A, Ascherman DP, et al. Required role of apoptotic myogenic precursors and toll-like receptor stimulation for the establishment of autoimmune myositis in experimental murine models［J］. Arthritis Rheumatol, 2015, 67(3): 809-822.

［6］Buckingham M, Relaix F. PAX3 and PAX7 as upstream regulators of myogenesis［J］. Semin Cell Dev Biol, 2015, 44: 115-125.

［7］Li P, Liu A, Liu C, et al. Role and mechanism of catechin in skeletal muscle cell differentiation［J］. J Nutr Biochem, 2019, 74: 108225.

［8］Xiang ChCh， Han F，Shi YX.Research on activating transcription factor 6 pathway and its related factors’ regulation on unfolded protein reaction in endoplasmic reticulum stress［J］. Acta Anatomica Sinica, 2015, 46(6): 857-861. (in Chinese)

向春晨，韩芳，石玉秀. 内质网应激中活化转录因子6通路及相关调控因子对未折叠蛋白反应的调控［J］. 解剖学报, 2015, 46(6): 857-861.

［9］De Rossi M, Bernasconi P, Baggi F, et al. Cytokines and chemokines are both expressed by human myoblasts: possible relevance for the immune pathogenesis of muscle inflammation［J］. Int Immunol, 2000, 12(9): 1329-1335.

［10］Ding M, Huang T, Zhu R, et al. Immunological behavior analysis of muscle cells under IFN-γ stimulation in vitro and in vivo［J］. Anat Rec (Hoboken), 2018, 301(9): 1551-1563.

［11］Wiendl H, Behrens L, Maier S, et al. Muscle fibers in inflammatory myopathies and cultured myoblasts express the nonclassical major histocompatibility antigen HLA-G［J］. Ann Neurol, 2000, 48(4): 679-684.

［12］Englund P, Lindroos E, Nennesmo I, et al. Skeletal muscle fibers express major histocompatibility complex class II antigens independently of inflammatory infiltrates in inflammatory myopathies［J］. Am J Pathol, 2001, 159(4): 1263-1273.

［13］Rayavarapu S, Coley W, Nagaraju K. Endoplasmic reticulum stress in skeletal muscle homeostasis and disease［J］. Curr Rheumatol Rep, 2012, 14(3): 238-243.

［14］Lightfoot AP, Nagaraju K, Mcardle A, et al. Understanding the origin of non-immune cell-mediated weakness in the idiopathic inflammatory myopathies-potential role of ER stress pathways［J］. Curr Opin Rheumatol, 2015, 27(6): 580-585.

［15］Deldicque L, Cani PD, Philp A, et al. The unfolded protein response is activated in skeletal muscle by high-fat feeding: potential role in the downregulation of protein synthesis［J］. Am J Physiol Endocrinol Metab, 2010, 299(5): E695-E705.

［16］Xiong G, Hindi SM, Mann AK, et al. The PERK arm of the unfolded protein response regulates satellite cell-mediated skeletal muscle regeneration［J］. Elife, 2017, 6: e2287.