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L-theanine induces skeletal muscle fiber type transformation by activation of prox1/CaN signaling pathway in C2C12 myotubes

  • Xiaoling Chen ORCID logo EMAIL logo , Man Zhang , Gang Jia , Hua Zhao , Guangmang Liu and Zhiqing Huang ORCID logo EMAIL logo
Published/Copyright: July 18, 2022

Abstract

The aim of this study was to investigate the effect and mechanism of L-theanine (LT) on muscle fiber type transformation in C2C12 myotubes. Our data showed that LT exhibited significantly higher slow oxidative muscle fiber expression and lower glycolytic fibers expression. In addition, LT significantly increased the activities of malate dehydrogenase (MDH) and succinic dehydrogenase (SDH), and decreased lactate dehydrogenase (LDH) activity, the calcineurin (CaN) activity and the protein expressions of nuclear factor of activated T cell 1 (NFATc1), prospero-related homeobox1 (prox1), and calcineurin A (CnA) were significantly increased. However, inhibition of CaN activity by cyclosporine A (CsA) abolished LT-induced increase of slow oxidative muscle fiber expression and decrease of glycolytic fibers expression. Moreover, inhibition of prox1 expression by prox1-siRNA disrupted LT-induced activation of CaN signaling pathway and muscle fiber type transformation. Taken together, these results indicated that LT could promote skeletal muscle fiber type transformation from type II to type I via activation of prox1/CaN signaling pathway.


Corresponding authors: Xiaoling Chen and Zhiqing Huang, Key Laboratory for Animal Disease-Resistance Nutrition of Chinese Ministry of Education, Institute of Animal Nutrition, Sichuan Agricultural University, Chengdu, Sichuan 611130, P.R. China, E-mail: ,

Funding source: Natural Science Foundation of Sichuan Province

Award Identifier / Grant number: 2022NSFSC0074

Funding source: Sichuan Agricultural University

Award Identifier / Grant number: 03570812

  1. Author contributions: X.L.C. and Z.Q.H. conceived and designed the experiments. M.Z performed the experiments and analyzed the data. X.L.C. wrote the paper. All authors read and approved the final manuscript.

  2. Research funding: Natural Science Foundation of Sichuan Province (2022NSFSC0074); Specific Research Supporting Program for Discipline Construction in Sichuan Agricultural University (03570812).

  3. Conflict of interest statement: The authors declare that they have no conflict of interests.

References

Chaillou, T. (2018). Skeletal muscle fiber type in hypoxia: adaptation to high-altitude exposure and under conditions of pathological hypoxia. Front. Physiol. 9: 1450, https://doi.org/10.3389/fphys.2018.01450.Search in Google Scholar

Chanda, M., Srikuea, R., Cherdchutam, W., Chairoungdua, A., and Piyachaturawat, P. (2016). Modulating effects of exercise training regimen on skeletal muscle properties in female polo ponies. BMC Vet. Res. 12: 245, https://doi.org/10.1186/s12917-016-0874-6.Search in Google Scholar

Chen, K., Yu, C.Y., Li, Z.H., Li, S., Yan, E.F., Song, Z.H., Zhang, H., Zhang, L.L., and Wang, T. (2020). Resveratrol improves meat quality, muscular antioxidant capacity, lipid metabolism and fiber type composition of intrauterine growth retarded pigs. Meat Sci. 170: 108237, https://doi.org/10.1016/j.meatsci.2020.108237.Search in Google Scholar

Chen, X.L., Zhang, M., Xue, Y.H., Liang, D.H., An, W.T., Jia, G., Zhao, H., Liu, G.M., and Huang, Z.Q. (2022). Effect of dietary L-theanine supplementation on skeletal muscle fiber type transformation in vivo. J. Nutr. Biochem. 99: 108859, https://doi.org/10.1016/j.jnutbio.2021.108859.Search in Google Scholar

Choi, Y.M. and Oh, H.K. (2016). Carcass performance, muscle fiber, meat quality, and sensory quality characteristics of crossbred pigs with different live weights. Korean J. Food Sci. Anim. Resour. 36: 389–396, https://doi.org/10.5851/kosfa.2016.36.3.389.Search in Google Scholar

Cooper, R. (2012). Green tea and theanine: health benefits. Int. J. Food Sci. Nutr. 63: 90–97, https://doi.org/10.3109/09637486.2011.629180.Search in Google Scholar

Dong, C., Zhang, X.Y., Liu, K.Q., Li, B.J., Chao, Z., Jiang, A.W., Li, R.Y., Li, P.H., Liu, H.L., and Wu, W.J. (2019). Comprehensive analysis of porcine prox1 gene and its relationship with meat quality traits. Animals 9: 744, https://doi.org/10.3390/ani9100744.Search in Google Scholar

Goldstein, J., Fletcher, S., Roth, E., Wu, C., Chun, A., and Horsley, V. (2014). Calcineurin/Nfatc1 signaling links skin stem cell quiescence to hormonal signaling during pregnancy and lactation. Genes Dev. 28: 983–994, https://doi.org/10.1101/gad.236554.113.Search in Google Scholar

Holloszy, J.O., Chen, M., Cartee, G.D., and Young, J.C. (1991). Skeletal muscle atrophy in old rats: differential changes in the three fiber types. Mech. Ageing Dev. 60: 199–213, https://doi.org/10.1016/0047-6374(91)90131-i.Search in Google Scholar

Hoppeler, H. (2016). Molecular networks in skeletal muscle plasticity. J. Exp. Biol. 219: 205–213, https://doi.org/10.1242/jeb.128207.Search in Google Scholar PubMed

Hou, Y., Su, L., Su, R., Luo, Y., Wang, B., Yao, D., Zhao, L., and Jin, Y. (2020). Effect of feeding regimen on meat quality, MyHC isoforms, AMPK, and PGC-1α genes expression in the biceps femoris muscle of Mongolia sheep. Food Sci. Nutr. 8: 2262–2270, https://doi.org/10.1002/fsn3.1494.Search in Google Scholar PubMed PubMed Central

Huang, Y.N., Xia, Q., Cui, Y.Y., Qu, Q.H., Wei, Y.M., and Jiang, Q.Y. (2020). Resveratrol increase the proportion of oxidative muscle fiber through the AdipoR1-AMPK-PGC-1α pathway in pigs. J. Funct. Foods 73: 104090, https://doi.org/10.1016/j.jff.2020.104090.Search in Google Scholar

Jia, A.F., Feng, J.H., Zhang, M.H., Chang, Y., Li, Z.Y., Hu, C.H., Zhen, L., Zhang, S.S., and Peng, Q.Q. (2015). Effects of immunological challenge induced by lipopolysaccharide on skeletal muscle fiber type conversion of piglets. J. Anim. Sci. 93: 5194–5203, https://doi.org/10.2527/jas.2015-9391.Search in Google Scholar PubMed

Kivelä, R., Salmela, I., Nguyen, Y.H., Petrova, T.V., Koistinen, H.A., Wiener, Z., and Alitalo, K. (2016). The transcription factor Prox1 is essential for satellite cell differentiation and muscle fibre-type regulation. Nat. Commun. 7: 13124, https://doi.org/10.1038/ncomms13124.Search in Google Scholar PubMed PubMed Central

Lee, L.S., Choi, J.H., Sung, M.J., Hur, J.M., Hur, H.J., Park, J.D., Kim, Y.C., Gu, E.J., Min, B.M., and Kim, H.J. (2015). Green tea changes serum and liver metabolomic profiles in mice with high-fat diet-induced obesity. Mol. Nutr. Food Res. 59: 784–794, https://doi.org/10.1002/mnfr.201400470.Search in Google Scholar PubMed

Li, B.J., Dong, C., Li, P.H., Ren, Z.Q., Wang, H., Yu, F.X., Ning, C.B., Liu, K.Q., Wei, W., Huang, R.H., et al.. (2016). Identification of candidate genes associated with porcine meat color traits by genome-wide transcriptome analysis. Sci. Rep. 6: 35224, https://doi.org/10.1038/srep35224.Search in Google Scholar PubMed PubMed Central

Livak, K.J. and Schmittgen, T.D. (2001). Analysis of relative gene expression data using real- time quantitative PCR and the 2−ΔΔCT method. Methods 25: 402–408, https://doi.org/10.1006/meth.2001.1262.Search in Google Scholar PubMed

Luo, P., Wang, L.N., Luo, L., Wang, L.S., Yang, K.L., Shu, G., Wang, S.B., Zhu, X.T., Gao, P., and Jiang, Q.Y. (2019). Ca2+-Calcineurin-NFAT pathway mediates the effect of thymol on oxidative metabolism and fiber-type switch in skeletal muscle. Food Funct. 10: 5166–5173, https://doi.org/10.1039/c8fo02248h.Search in Google Scholar PubMed

Martins, K.J.B., St-Louis, M., Murdoch, G.K., MacLean, I.M., McDonald, P., Dixon, W.T., Putman, T., and Michel, R.N. (2012). Nitric oxide synthase inhibition prevents activity-induced calcineurin–NFATc1 signaling and fast-to-slow skeletal muscle fibre type conversions. J. Physiol. 590: 1427–1442, https://doi.org/10.1113/jphysiol.2011.223370.Search in Google Scholar PubMed PubMed Central

McCullagh, K.J., Calabria, E., Pallafacchina, G., Ciciliot, S., Serrano, A.L., Argentini, C., Kalhovde, J.M., Lømo, T., and Schiaffino, S. (2004). NFAT is a nerve activity sensor in skeletal muscle and controls activity-dependent myosin switching. Proc. Natl. Acad. Sci. U.S.A. 101: 10590–10595, https://doi.org/10.1073/pnas.0308035101.Search in Google Scholar PubMed PubMed Central

Naya, F.J., Mercer, B., Shelton, J., Richardson, J.A., Williams, R.S., and Olson, E.N. (2000). Stimulation of slow skeletal muscle fiber gene expression by calcineurin in vivo. J. Biol. Chem. 275: 4545–4548, https://doi.org/10.1074/jbc.275.7.4545.Search in Google Scholar PubMed

Ohno, Y., Yamada, S., Goto, A., Ikuta, A., Sugiura, T., Ohira, Y., Yoshioka, T., and Goto, K. (2012). Effects of heat stress on muscle mass and the expression levels of heat shock proteins and Lysosomal cathepsin L. in soleus muscle of young and aged mice. Mol. Cell. Biochem. 369: 45–53, doi:https://doi.org/10.1007/s11010-012-1367-y.Search in Google Scholar PubMed

Peng, W.Q., Xiao, G., Li, B.Y., Guo, Y.Y., Guo, L., and Tang, Q.Q. (2021). l-Theanine activates the browning of white adipose tissue through the AMPK/α-ketoglutarate/prdm16 axis and ameliorates diet-induced obesity in mice. Diabetes 70: 1485–1472, https://doi.org/10.2337/db20-1210.Search in Google Scholar PubMed

Petchey, L.K., Risebro, C.A., Vieira, J.M., Roberts, T., Bryson, J.B., Greensmith, L., Lythgoe, M.F., and Riley, P.R. (2014). Loss of Prox1 in striated muscle causes slow to fast skeletal muscle fiber conversion and dilated cardiomyopathy. Proc. Natl. Acad. Sci. U.S.A. 111: 9515–9520, https://doi.org/10.1073/pnas.1406191111.Search in Google Scholar PubMed PubMed Central

Ryu, Y.C. and Kim, B.C. (2005). The relationship between muscle fiber characteristics, postmortem metabolic rate, and meat quality of pig longissimus dorsi muscle. Meat Sci. 71: 351–357, https://doi.org/10.1016/j.meatsci.2005.04.015.Search in Google Scholar PubMed

Schiaffino, S. and Reggiani, C. (2011). Fiber types in mammalian skeletal muscles. Physiol. Rev. 91: 1447–1531, https://doi.org/10.1152/physrev.00031.2010.Search in Google Scholar PubMed

Talbot, J. and Marves, L. (2016). Skeletal muscle fiber type: using insights from muscle developmental biology to dissect targets for susceptibility and resistance to muscle disease. Wiley Interdiscip. Rev. Dev. Biol. 5: 518–534, https://doi.org/10.1002/wdev.230.Search in Google Scholar PubMed PubMed Central

Türközü, D. and Sanlier, N. (2017). L-theanine, unique amino acid of tea, and its metabolism, health effects, and safety. Crit. Rev. Food Sci. Nutr. 57: 1681–1687.10.1080/10408398.2015.1016141Search in Google Scholar PubMed

Wang, J., Chen, T., Feng, F., Wei, H., Pang, W.J., Yang, G.S., and Shen, Q.W. (2014). KLF15 regulates slow myosin heavy chain expression through NFATc1 in C2C12 myotubes. Biochem. Biophys. Res. Commun. 446: 1231–1236, https://doi.org/10.1016/j.bbrc.2014.03.091.Search in Google Scholar PubMed

Wang, L.N., Wang, Z., Kang, K.L., Shu, G., Wang, S.B., Gao, P., Zhu, X.T., Xi, Q.Y., Zhang, Y.L., and Jiang, Q.Y. (2016). Epigallocatechin gallate reduces slow-twitch muscle fiber formation and mitochondrial biosynthesis in C2C12 Cells by repressing AMPK activity and PGC-1α expression. J. Agric. Food Chem. 64: 6517–6523, https://doi.org/10.1021/acs.jafc.6b02193.Search in Google Scholar PubMed

Xu, M., Chen, X.L., Huang, Z.Q., Chen, D.W., Li, M.Z., He, J., Chen, H., Zheng, P., Yu, J., Luo, Y.H., et al.. (2021). Effects of dietary grape seed proanthocyanidin extract supplementation on meat quality, muscle fiber characteristics and antioxidant capacity of finishing pigs. Food Chem. 367: 130781, https://doi.org/10.1016/j.foodchem.2021.130781.Search in Google Scholar PubMed

Received: 2022-04-24
Accepted: 2022-06-28
Published Online: 2022-07-18
Published in Print: 2022-09-27

© 2022 Walter de Gruyter GmbH, Berlin/Boston

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