Neuregulin-1β attenuates sepsis-induced diaphragm atrophy by activating the PI3K/Akt signaling pathway

  • Jin Wu
  • Hua Liu
  • Ting Chu
  • Peng JiangEmail author
  • Shi-tong LiEmail author
Original Article


The aim of this study was to investigate the protective effects of neuregulin-1β (NRG-1β) on sepsis-induced diaphragm atrophy and the possible underlying mechanisms. Sprague–Dawley rats were randomly divided into sham, sepsis and NRG groups. Sepsis was induced by cecal ligation and puncture (CLP). In the NRG group, rats received tail vein injections of NRG-1β (10 μg/kg) every 12 h for 72 h after CLP. At 3 days after surgery, diaphragm contractile forces were measured by determining the force-frequency curve and muscle fiber areas by hematoxylin–eosin staining. Moreover, the NRG-1 expression level in the diaphragm was detected by Western blotting. Furthermore, the proteins in the PI3K/Akt signaling pathway and its downstream Akt-mTOR and Akt-FOXO axes were detected by Western blotting analysis. In L6 myotubes treated with lipopolysaccharide (LPS) and NRG-1β, PI3K/Akt signaling pathway-related protein expression was further determined using the PI3K inhibitor LY294002. Exogenous NRG-1β could compensate for sepsis-induced diminished NRG-1 in the diaphragm and attenuate the reduction in diaphragm contractile forces and muscle fiber areas during sepsis. Moreover, NRG-1β treatment could activate the PI3K/Akt signaling pathway in the diaphragm during sepsis. The inhibition of p70S6K and 4E-BP1 on the Akt-mTOR axis and the increased expression of Murf1 on the Akt-FOXO axis were reversed after NRG-1 treatment. In addition, NRG-1β could activate the PI3K/Akt signaling pathway in L6 myotubes treated with LPS, while the PI3K inhibitor LY294002 blocked the effects of NRG-1β. NRG-1 expression in the diaphragm was reduced during sepsis, and exogenously administered recombinant human NRG-1β could attenuate sepsis-induced diaphragm atrophy by activating the PI3K/Akt signaling pathway.


Sepsis Neuregulin-1 PI3K/Akt pathway Diaphragm Atrophy 



This study was supported by the Introduction Program of High-Level Innovation and Entrepreneurship Talents in Jiangsu Province (2018) and the Scientific Research Foundation of Affiliated Hospital of Jiangsu University (jdfyRC2017008).

Compliance with ethical standards

Conflicts of interest

Jin Wu, Hua Liu, Ting Chu, Peng Jiang and Shi-tong Li declare that they have no conflict of interest.


  1. Aedo JE, Reyes AE, Avendaño-Herrera R, Molina A, Valdés JA (2015) Bacterial lipopolysaccharide induces rainbow trout myotube atrophy via Akt/FoxO1/Atrogin-1 signaling pathway. Acta Biochim Biophys Sin (Shanghai) 47:932–937. CrossRefGoogle Scholar
  2. An T, Zhang Y, Huang Y, Zhang R, Yin S, Guo X, Wang Y, Zou C, Wei B, Lv R, Zhou Q, Zhang J (2013) Neuregulin-1 protects against doxorubicin-induced apoptosis in cardiomyocytes through an Akt-dependent pathway. Physiol Res 62:379–385Google Scholar
  3. Cai MX, Shi XC, Chen T, Tan ZN, Lin QQ, Du SJ, Tian ZJ (2016) Exercise training activates neuregulin 1/ErbB signaling and promotes cardiac repair in a rat myocardial infarction model. Life Sci 149:1–9. CrossRefGoogle Scholar
  4. Callahan LA, Supinski GS (2009) Sepsis-induced myopathy. Crit Care Med 37:S354–S367. CrossRefGoogle Scholar
  5. Chen W, Liu Y, Xue G, Zhang L, Zhang L, Shao S (2016) Diazoxide protects L6 skeletal myoblasts from H2O2-induced apoptosis via the phosphatidylinositol-3 kinase/Akt pathway. Inflamm Res 65:53–60. CrossRefGoogle Scholar
  6. Constantin D, McCullough J, Mahajan RP, Greenhaff PL (2011) Novel events in the molecular regulation of muscle mass in critically ill patients. J Physiol 589:3883–3895. CrossRefGoogle Scholar
  7. Crossland H, Constantin-Teodosiu D, Gardiner SM, Constantin D, Greenhaff PL (2008) A potential role for Akt/FOXO signalling in both protein loss and the impairment of muscle carbohydrate oxidation during sepsis in rodent skeletal muscle. J Physiol 586:5589–5600. CrossRefGoogle Scholar
  8. Falls DL (2003) Neuregulins: functions, forms, and signaling strategies. Exp Cell Res 284:14–30.,00102-7 CrossRefGoogle Scholar
  9. Fang SJ, Wu XS, Han ZH, Zhang XX, Wang CM, Li XY, Lu LQ, Zhang JL (2010) Neuregulin-1 preconditioning protects the heart against ischemia/reperfusion injury through a PI3K/Akt-dependent mechanism. Chin Med J (Engl) 123:3597–3604. Google Scholar
  10. Fleischmann C, Scherag A, Adhikari NK, Hartog CS, Tsaganos T, Schlattmann P, Angus DC, Reinhart K, International Forum of Acute Care Trialists (2016) International forum of acute care trialists. Am J Respir Crit Care Med 193:259–272. CrossRefGoogle Scholar
  11. Glass DJ (2010) PI3 kinase regulation of skeletal muscle hypertrophy and atrophy. Curr Top Microbiol Immunol 346:267–278. Google Scholar
  12. Gordon BS, Kelleher AR, Kimball SR (2013) Regulation of muscle protein synthesis and the effects of catabolic states. Int J Biochem Cell Biol 45:2147–2157. CrossRefGoogle Scholar
  13. Hellyer NJ, Kim MS, Koland JG (2001) Heregulin-dependent activation of phosphoinositide 3-kinase and Akt via the ErbB2/ErbB3 co-receptor. J Biol Chem 276:42153–42161. CrossRefGoogle Scholar
  14. Hellyer NJ, Mantilla CB, Park EW, Zhan WZ, Sieck GC (2006) Neuregulin-dependent protein synthesis in C2C12 myotubes and rat diaphragm muscle. Am J Physiol Cell Physiol 291:C1056–C1061. CrossRefGoogle Scholar
  15. Jagoe RT, Goldberg AL (2001) What do we really know about the ubiquitin-proteasome pathway in muscle atrophy? Curr Opin Clin Nutr Metab Care 4:183–190CrossRefGoogle Scholar
  16. Jiao G, Hao L, Wang M, Zhong B, Yu M, Zhao S, Wang P, Feng R, Tan S, Chen L (2017) Upregulation of endoplasmic reticulum stress is associated with diaphragm contractile dysfunction in a rat model of sepsis. Mol Med Rep 15:366–374. CrossRefGoogle Scholar
  17. Jie B, Zhang X, Wu X, Xin Y, Liu Y, Guo Y (2012) Neuregulin-1 suppresses cardiomyocyte apoptosis by activating PI3K/Akt and inhibiting mitochondrial permeability transition pore. Mol Cell Biochem 370:35–43. CrossRefGoogle Scholar
  18. Jung B, Nougaret S, Conseil M, Coisel Y, Futier E, Chanques G, Molinari N, Lacampagne A, Matecki S, Jaber S (2014) Sepsis is associated with a preferential diaphragmatic atrophy: a critically ill patient study using tridimensional computed tomography. Anesthesiology 120:1182–1191. CrossRefGoogle Scholar
  19. Kazi AA, Pruznak AM, Frost RA, Lang CH (2011) Sepsis-induced alterations in protein-protein interactions within mTOR complex 1 and the modulating effect of leucine on muscle protein synthesis. Shock 35:117–125. CrossRefGoogle Scholar
  20. Kim JA, Park HS, Park KI, Hong GE, Nagappan A, Zhang J, Han DY, Shin SC, Won CG, Kim EH, Kim GS (2013) Proteome analysis of the anti-inflammatory response of flavonoids isolated from Korean Citrus aurantium L. in lipopolysaccharide-induced L6 rat skeletal muscle cells. Am J Chin Med 41:901–912. CrossRefGoogle Scholar
  21. Lebrasseur NK, Coté GM, Miller TA, Fielding RA, Sawyer DB (2003) Regulation of neuregulin/ErbB signaling by contractile activity in skeletal muscle. Am J Physiol Cell Physiol 284:C1149–C1155. CrossRefGoogle Scholar
  22. Liu L, Xie F, Wei K, Hao XC, Li P, Cao J, Min S (2016) Sepsis induced denervation-like changes at the neuromuscular junction. J Surg Res 200:523–532. CrossRefGoogle Scholar
  23. Maes K, Stamiris A, Thomas D, Cielen N, Smuder A, Powers SK, Leite FS, Hermans G, Decramer M, Hussain SN, Gayan-Ramirez G (2014) Effects of controlled mechanical ventilation on sepsis-induced diaphragm dysfunction in rats. Crit Care Med 42:e772–e782. CrossRefGoogle Scholar
  24. Monahan LJ (2013) Acute respiratory distress syndrome. Curr Probl Pediatr Adolesc Health Care 43:278–284. CrossRefGoogle Scholar
  25. Rimer M (2007) Neuregulins at the neuromuscular synapse: past, present, and future. J Neurosci Res 85:1827–1833. CrossRefGoogle Scholar
  26. Sandri M (2008) Signaling in muscle atrophy and hypertrophy. Physiology (Bethesda) 23:160–170. Google Scholar
  27. Seethala RR, Hou PC, Aisiku IP, Frendl G, Park PK, Mikkelsen ME, Chang SY, Gajic O, Sevransky J (2017) Early risk factors and the role of fluid administration in developing acute respiratory distress syndrome in septic patients. Ann Intensive Care 7:11. CrossRefGoogle Scholar
  28. Shiota C, Abe T, Kawai N, Ohno A, Teshima-Kondo S, Mori H, Terao J, Tanaka E, Nikawa T (2015) flavones Inhibit LPS-induced atrogin-1/MAFbx expression in mouse C2C12 skeletal myotubes. J Nutr Sci Vitaminol (Tokyo) 61:188–194. CrossRefGoogle Scholar
  29. Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, Bauer M, Bellomo R, Bernard GR, Chiche JD, Coopersmith CM, Hotchkiss RS, Levy MM, Marshall JC, Martin GS, Opal SM, Rubenfeld GD, van der Poll T, Vincent JL, Angus DC (2016) The third international consensus definitions for sepsis and septic shock (sepsis-3). JAMA 315:801–810. CrossRefGoogle Scholar
  30. Stana F, Vujovic M, Mayaki D, Leduc-Gaudet JP, Leblanc P, Huck L, Hussain SNA (2017) Differential regulation of the autophagy and proteasome pathways in skeletal muscles in sepsis. Crit Care Med 45:e971–e979. CrossRefGoogle Scholar
  31. Supinski GS, Vanags J, Callahan LA (2009) Effect of proteasome inhibitors on endotoxin-induced diaphragm dysfunction. Am J Physiol Lung Cell Mol Physiol 296:L994–L1001. CrossRefGoogle Scholar
  32. Svanberg E, Frost RA, Lang CH, Isgaard J, Jefferson LS, Kimball SR, Vary TC (2000) IGF-I/IGFBP-3 binary complex modulates sepsis-induced inhibition of protein synthesis in skeletal muscle. Am J Physiol Endocrinol Metab 279:E1145–E1158. CrossRefGoogle Scholar
  33. Wang MM, Hao LY, Guo F, Zhong B, Zhong XM, Yuan J, Hao YF, Zhao S, Sun XF, Lei M, Jiao GY (2017) Decreased intracellular [Ca2+] coincides with reduced expression of Dhprα1s, RyR1, and diaphragmatic dysfunction in a rat model of sepsis. Muscle Nerve 56:1128–1136. CrossRefGoogle Scholar
  34. Wu J, Li ST (2015) Dexmedetomidine may produce extra protective effects on sepsis-induced diaphragm injury. Chin Med J (Engl) 128:1407–1411. CrossRefGoogle Scholar
  35. Wu J, Zhang JY, Gong Y, Li ST (2016) Slowed relaxation of diaphragm in septic rats is associated with reduced expression of sarco-endoplasmic reticulum Ca2+-ATPase genes SERCA1 and SERCA2. Muscle Nerve 54:1108–1113. CrossRefGoogle Scholar
  36. Wu J, Jin T, Wang H, Li ST (2017) Sepsis decreases the activity of acetylcholinesterase by reducing its expression at the neuromuscular junction. Mol Med Rep 16:5263–5268. CrossRefGoogle Scholar
  37. Xie F, Min S, Chen J, Yang J, Wang X (2017) Ulinastatin inhibited sepsis-induced spinal inflammation to alleviate peripheral neuromuscular dysfunction in an experimental rat model of neuromyopathy. J Neurochem 143:225–235. CrossRefGoogle Scholar
  38. Zhou Q, Pan X, Wang L, Wang X, Xiong D (2016) The protective role of neuregulin-1: a potential therapy for sepsis-induced cardiomyopathy. Eur J Pharmacol 788:234–240. CrossRefGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of AnesthesiologyAffiliated Hospital of Jiangsu UniversityZhenjiangChina
  2. 2.Department of StomatologyAffiliated People’s Hospital of Jiangsu UniversityZhenjiangChina
  3. 3.Department of Anesthesiology, Shanghai General HospitalShanghai Jiao Tong University School of MedicineShanghaiChina
  4. 4.Department of Anesthesiology, The Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

Personalised recommendations