Skeletal Muscle Atrophy Induced by Diabetes Is Mediated by Non-Selective Channels and Prevented by Boldine
Luis A Cea, Walter Vásquez, Romina Hernández-Salinas, Alejandra Z Vielma, Mario Castillo-Ruiz, Victoria Velarde, Magdiel Salgado, Juan C Sáez, Luis A Cea, Walter Vásquez, Romina Hernández-Salinas, Alejandra Z Vielma, Mario Castillo-Ruiz, Victoria Velarde, Magdiel Salgado, Juan C Sáez
Abstract
Individuals with diabetes mellitus present a skeletal muscle myopathy characterized by atrophy. However, the mechanism underlying this muscular alteration remains elusive, which makes it difficult to design a rational treatment that could avoid the negative consequences in muscles due to diabetes. In the present work, the atrophy of skeletal myofibers from streptozotocin-induced diabetic rats was prevented with boldine, suggesting that non-selective channels inhibited by this alkaloid are involved in this process, as has previously shown for other muscular pathologies. Accordingly, we found a relevant increase in sarcolemma permeability of skeletal myofibers of diabetic animals in vivo and in vitro due to de novo expression of functional connexin hemichannels (Cx HCs) containing connexins (Cxs) 39, 43, and 45. These cells also expressed P2X7 receptors, and their inhibition in vitro drastically reduced sarcolemma permeability, suggesting their participation in the activation of Cx HCs. Notably, sarcolemma permeability of skeletal myofibers was prevented by boldine treatment that blocks Cx43 and Cx45 HCs, and now we demonstrated that it also blocks P2X7 receptors. In addition, the skeletal muscle alterations described above were not observed in diabetic mice with myofibers deficient in Cx43/Cx45 expression. Moreover, murine myofibers cultured for 24 h in high glucose presented a drastic increase in sarcolemma permeability and levels of NLRP3, a molecular member of the inflammasome, a response that was also prevented by boldine, suggesting that, in addition to the systemic inflammatory response found in diabetes, high glucose can promote the expression of functional Cx HCs and activation of the inflammasome in skeletal myofibers. Therefore, Cx43 and Cx45 HCs play a critical role in myofiber degeneration, and boldine could be considered a potential therapeutic agent to treat muscular complications due to diabetes.
Keywords: calcium atrophy; connexins; hemichannel blocker; sarcolemma permeability.
Conflict of interest statement
The authors declare no conflict of interest.
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References
- Biondi-Zoccai G.G., Abbate A., Liuzzo G., Biasucci L.M. Atherothrombosis, inflammation, and diabetes. J. Am. Coll. Cardiol. 2003;41:1071–1077. doi: 10.1016/S0735-1097(03)00088-3.
- Forbes J.M., Cooper M.E. Mechanisms of diabetic complications. Physiol. Rev. 2013;93:137–188. doi: 10.1152/physrev.00045.2011.
- DeFronzo R.A. Glucose intolerance and aging. Diabetes Care. 1981;4:493–501. doi: 10.2337/diacare.4.4.493.
- D’Souza D.M., Al-Sajee D., Hawke T.J. Diabetic myopathy: Impact of diabetes mellitus on skeletal muscle progenitor cells. Front. Physiol. 2013;4:379. doi: 10.3389/fphys.2013.00379.
- Cea L.A., Cisterna B.A., Puebla C., Frank M., Figueroa X.F., Cardozo C., Willecke K., Latorre R., Sáez J.C. De novo expression of connexin hemichannels in denervated fast skeletal muscles leads to atrophy. Proc. Natl. Acad. Sci. USA. 2013;110:16229–16234. doi: 10.1073/pnas.1312331110.
- Cea L.A., Balboa E., Vargas A.A., Puebla C., Brañes M.C., Escamilla R., Regueira T., Sáez J.C. De novo expression of functional connexins 43 and 45 hemichannels increases sarcolemmal permeability of skeletal myofibers during endotoxemia. Biochim. Biophys. Acta Mol. Basis Dis. 2019;1865:2765–2773. doi: 10.1016/j.bbadis.2019.06.014.
- Cisterna B.A., Vargas A.A., Puebla C., Fernández P., Escamilla R., Lagos C.F., Matus M.F., Vilos C., Cea L.A., Barnafi E., et al. Active acetylcholine receptors prevent the atrophy of skeletal muscles and favor reinnervation. Nat. Commun. 2020;11:1073. doi: 10.1038/s41467-019-14063-8.
- Balboa E., Saavedra-Leiva F., Cea L.A., Vargas A.A., Ramírez V., Escamilla R., Sáez J.C., Regueira T. Sepsis-Induced Channelopathy in Skeletal Muscles is Associated with Expression of Non-Selective Channels. Shock. 2018;49:221–228. doi: 10.1097/SHK.0000000000000916.
- Peng B., Xu C., Wang S., Zhang Y., Li W. The Role of Connexin Hemichannels in Inflammatory Diseases. Biology. 2022;11:237. doi: 10.3390/biology11020237.
- Donath M.Y., Dinarello C.A., Mandrup-Poulsen T. Targeting innate immune mediators in type 1 and type 2 diabetes. Nat. Rev. Immunol. 2019;19:734–746. doi: 10.1038/s41577-019-0213-9.
- Hernández-Salinas R., Vielma A.Z., Arismendi M.N., Boric M.P., Sáez J.C., Velarde V. Boldine prevents renal alterations in diabetic rats. J. Diabetes Res. 2013;2013:593672. doi: 10.1155/2013/593672.
- Yi C., Ezan P., Fernández P., Schmitt J., Sáez J.C., Giaume C., Koulakoff A. Inhibition of glial hemichannels by boldine treatment reduces neuronal suffering in a murine model of Alzheimer’s disease. Glia. 2017;65:1607–1625. doi: 10.1002/glia.23182.
- Koshimizu T., Koshimizu M., Stojilkovic S.S. Contributions of the C-terminal domain to the control of P2X receptor desensitization. J. Biol. Chem. 1999;274:37651–37657. doi: 10.1074/jbc.274.53.37651.
- Choi E.J., Palacios-Prado N., Sáez J.C., Lee J. Identification of Cx45 as a Major Component of GJs in HeLa Cells. Biomolecules. 2020. 10:1389.
- Cea L.A., Puebla C., Cisterna B.A., Escamilla R., Vargas A.A., Frank M., Martínez-Montero P., Prior C., Molano J., Esteban-Rodríguez I., et al. Fast skeletal myofibers of mdx mouse, model of Duchenne muscular dystrophy, express connexin hemichannels that lead to apoptosis. Cell. Mol. Life Sci. 2016;73:2583–2599. doi: 10.1007/s00018-016-2132-2.
- Cea L.A., Fernández G., Arias-Bravo G., Castillo-Ruiz M., Escamilla R., Brañes M.C., Sáez J.C. Blockade of Hemichannels Normalizes the Differentiation Fate of Myoblasts and Features of Skeletal Muscles from Dysferlin-Deficient Mice. Int. J. Mol. Sci. 2020;21:6025. doi: 10.3390/ijms21176025.
- Messemer N., Kunert C., Grohmann M., Sobottka H., Nieber K., Zimmermann H., Franke H., Nörenberg W., Straub I., Schaefer M., et al. P2X7 receptors at adult neural progenitor cells of the mouse subventricular zone. Neuropharmacology. 2013;73:122–137. doi: 10.1016/j.neuropharm.2013.05.017.
- Allsopp R.C., Dayl S., Dayel A.B., Schmid R., Evans R.J. Mapping the Allosteric Action of Antagonists A740003 and A438079 Reveals a Role for the Left Flipper in Ligand Sensitivity at P2X7 Receptors. Mol. Pharmacol. 2018;93:553–562. doi: 10.1124/mol.117.111021.
- Cline G.W., Petersen K.F., Krssak M., Shen J., Hundal R.S., Trajanoski Z., Inzucchi S., Dresner A., Rothman D.L., Shulman G.I. Impaired glucose transport as a cause of decreased insulin-stimulated muscle glycogen synthesis in type 2 diabetes. New Engl. J. Med. 1999;341:240–246. doi: 10.1056/NEJM199907223410404.
- Sáez J.C., Contreras-Duarte S., Labra V.C., Santibañez C.A., Mellado L.A., Inostroza C.A., Alvear T.F., Retamal M.A., Velarde V., Orellana J.A. Interferon-γ and high glucose-induced opening of Cx43 hemichannels causes endothelial cell dysfunction and damage. Biochim. Biophys. Acta Mol. Cell Res. 2020;1867:118720. doi: 10.1016/j.bbamcr.2020.118720.
- Schalper KA, Sánchez HA, Lee SC, Altenberg GA, Nathanson MH, Sáez JC. Connexin 43 hemichannels mediate the Ca2+ influx induced by extracellular J Physiol Cell Physiol. 2010;299:C1504–C1515.
- Liang X., Samways D.S.K., Wolf K., Bowles E.A., Richards J.P., Bruno J., Dutertre S., DiPaolo R.J., Egan T.M. Quantifying Ca2+ Current and Permeability in ATP-gated P2X7 Receptors. J. Biol. Chem. 2015;290:7930–7942. doi: 10.1074/jbc.M114.627810.
- Balboa E., Saavedra F., Cea L.A., Ramírez V., Escamilla R., Vargas A.A., Regueira T., Sáez J.C. Vitamin E Blocks Connexin Hemichannels and Prevents Deleterious Effects of Glucocorticoid Treatment on Skeletal Muscles. Int. J. Mol. Sci. 2020;21:4094. doi: 10.3390/ijms21114094.
- Wang N., De Vuyst E., Ponsaerts R., Boengler K., Palacios-Prado N., Wauman J., Lai C.P., De Bock M., Decrock E., Bol M., et al. Selective inhibition of Cx43 hemichannels by Gap19 and its impact on myocardial ischemia/reperfusion injury. Basic Res. Cardiol. 2013;108:309. doi: 10.1007/s00395-012-0309-x.
- Rusiecka O.M., Tournier M., Molica F., Kwak B.R. Pannexin1 channels—A potential therapeutic target in inflammation. Front. Cell Dev. Biol. 2022;10:1020826. doi: 10.3389/fcell.2022.1020826.
- Solini A., Novak I. Role of the P2X7 receptor in the pathogenesis of type 2 diabetes and its microvascular complications. Curr. Opin. Pharmacol. 2019;47:75–81. doi: 10.1016/j.coph.2019.02.009.
- Giaume C., Naus C.C., Sáez J.C., Leybaert L. Glial Connexins and Pannexins in the Healthy and Diseased Brain. Physiol. Rev. 2021;101:93–145. doi: 10.1152/physrev.00043.2018.
- De Vuyst E., Wang N., Decrock E., De Bock M., Vinken M., Van Moorhem M., Lai C., Culot M., Rogiers V., Cecchelli R., et al. Ca2+ regulation f connexin 43 hemichannels in C6 glioma and glial cells. Cell Calcium. 2009;46:176–187. doi: 10.1016/j.ceca.2009.07.002.
- Araya R., Eckardt D., Maxeiner S., Krüger O., Theis M., Willecke K., Sáez J.C. Expression of connexins during differentiation and regeneration of skeletal muscle: Functional relevance of connexin43. J. Cell Sci. 2005;118:27–37. doi: 10.1242/jcs.01553.
- Anderson C., Catoe H., Werner R. MIR-206 regulates connexin43 expression during skeletal muscle development. Nucleic Acids Res. 2006;34:5863–5871. doi: 10.1093/nar/gkl743.
- Cea L.A., Balboa E., Puebla C., Vargas A.A., Cisterna B.A., Escamilla R., Regueira T., Sáez J.C. Dexamethasone-induced muscular atrophy is mediated by functional expression of connexin-based hemichannels. Biochim. Biophys. Acta. 2016;1862:1891–1899. doi: 10.1016/j.bbadis.2016.07.003.
- Fernández G., Arias-Bravo G., Bevilacqua J.A., Castillo-Ruiz M., Caviedes P., Sáez J.C., Cea L.A. Myofibers deficient in connexins 43 and 45 expression protect mice from skeletal muscle and systemic dysfunction promoted by a dysferlin mutation. Biochim. Biophys. Acta Mol. Basis Dis. 2020;1866:165800. doi: 10.1016/j.bbadis.2020.165800.
- Kimura M., Kimura I., Nakamura T., Nojima H. Diabetic state-induced modification of resting membrane potential and conductance in diaphragm muscle of alloxan and diabetic KK-CAy mice. Diabetologia. 1988;31:103–107. doi: 10.1007/BF00395556.
- Hernández-Ochoa E.O., Banks Q., Schneider M.F. Acute Elevated Glucose Promotes Abnormal Action Potential-Induced Ca2+ Transients in Cultured Skeletal Muscle Fibers. J. Diabetes Res. 2017;2017:1509048. doi: 10.1155/2017/1509048.
- Retamal M.A., Cortés C.J., Reuss L., Bennett M.V., Sáez J.C. S-nitrosylation and permeation through connexin 43 hemichannels in astrocytes: Induction by oxidant stress and reversal by reducing agents. Proc. Natl. Acad. Sci. USA. 2006;103:4475–4480. doi: 10.1073/pnas.0511118103.
- Retamal M.A., Schalper K.A., Shoji K.F., Bennett M.V., Sáez J.C. Opening of connexin 43 hemichannels is increased by lowering intracellular redox potential. Proc. Natl. Acad. Sci. USA. 2007;104:8322–8327. doi: 10.1073/pnas.0702456104.
- Figueroa X.F., Lillo M.A., Gaete P.S., Riquelme M.A., Sáez J.C. Diffusion of nitric oxide across cell membranes of the vascular wall requires specific connexin-based channels. Neuropharmacology. 2013;75:471–478. doi: 10.1016/j.neuropharm.2013.02.022.
- Orellana J.A., Díaz E., Schalper K.A., Vargas A.A., Bennett M.V., Sáez J.C. Cation permeation through connexin 43 hemichannels is cooperative, competitive and saturable with parameters depending on the permeant species. Biochem. Biophys. Res. Commun. 2011;409:603–609. doi: 10.1016/j.bbrc.2011.05.031.
- Schalper K.A., Palacios-Prado N., Orellana J.A., Sáez J.C. Currently used methods for identification and characterization of hemichannels. Cell Commun. Adhes. 2008;15:207–218. doi: 10.1080/15419060802014198.
- Lee H.M., Kim J.J., Kim H.J., Shong M., Ku B.J., Jo E.K. Upregulated NLRP3 inflammasome activation in patients with type 2 diabetes. Diabetes. 2013;62:194–204. doi: 10.2337/db12-0420.
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