title: Altered exosomal protein expression in the serum of NF-κB knockout mice following skeletal muscle ischemia-reperfusion injury.
abstract: BACKGROUND
  The NF-κB signaling pathway plays a role in local and remote tissue damage following ischemia-reperfusion (I/R) injury to skeletal muscles. Evidence suggests that exosomes can act as intercellular communicators by transporting active proteins to remote cells and may play a role in regulating inflammatory processes. This study aimed to profile the exosomal protein expression in the serum of NF-κB knockout mice following skeletal muscle ischemia-reperfusion injury.
  
  
  RESULTS
  To investigate the potential changes in protein expression mediated by NF-κB in secreted exosomes in the serum following I/R injury, the levels of circulating exosomal proteomes in C57BL/6 and NF-κB(-/-) mice were compared using two dimensional differential in-gel electrophoresis (2-DE), liquid chromatography tandem mass spectrometry (LC-MS/MS), and proteomic analysis. In C57BL/6 mice, the levels of circulating exosomal proteins, including complement component C3 prepropeptide, PK-120 precursor, alpha-amylase one precursor, beta-enolase isoform 1, and adenylosuccinate synthetase isozyme 1, increased following I/R injury. However, in the NF-κB(-/-) mice, the expression of the following was upregulated in the exosomes: protease, serine 1; glyceraldehyde-3-phosphate dehydrogenase-like isoform 1; glyceraldehyde-3-phosphate dehydrogenase; and pregnancy zone protein. In contrast, the expression of apolipoprotein B, complement component C3 prepropeptide, and immunoglobulin kappa light chain variable region was downregulated in NF-κB(-/-) mice. Bioinformatic annotation using the Protein Analysis Through Evolutionary Relationships (PANTHER) database revealed that the expression of the exosomal proteins that participate in metabolic processes and in biological regulation was lower in NF-κB(-/-) mice than in C57BL/6 mice, whereas the expression of proteins that participate in the response to stimuli, in cellular processes, and in the immune system was higher.
  
  
  CONCLUSIONS
  The data presented in this study suggest that NF-κB might regulate exosomal protein expression at a remote site via circulation following I/R injury.
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. johnson.c.yang@gmail.com.
authors.forename: Johnson Chia-Shen
authors.initials: JC
authors.lastname: Yang
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. tanukikimo@yahoo.com.tw.
authors.forename: Ming-Wei
authors.initials: MW
authors.lastname: Lin
authors.affiliation: Department of Neurosurgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, Kaohsiung City, Taiwan. ersh2127@adm.cgmh.org.tw.
authors.forename: Cheng-Shyuan
authors.initials: CS
authors.lastname: Rau
authors.affiliation: Department of Plastic Surgery, E-Da Hospital, I-Shou University, Kaohsiung City, Taiwan. jengfamily@hotmail.com.
authors.forename: Seng-Feng
authors.initials: SF
authors.lastname: Jeng
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. rabbit670326@yahoo.com.tw.
authors.forename: Tsu-Hsiang
authors.initials: TH
authors.lastname: Lu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. rabbit670326@yahoo.com.tw.
authors.forename: Yi-Chan
authors.initials: YC
authors.lastname: Wu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. libe320@yahoo.com.tw.
authors.forename: Yi-Chun
authors.initials: YC
authors.lastname: Chen
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. showlingltzeng@yahoo.com.tw.
authors.forename: Siou-Ling
authors.initials: SL
authors.lastname: Tzeng
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. alice8818@yahoo.com.tw.
authors.forename: Chia-Jung
authors.initials: CJ
authors.lastname: Wu
authors.affiliation: Department of Plastic and Reconstructive Surgery, Kaohsiung Chang Gung Memorial Hospital and Chang Gung University College of Medicine, No. 123, Ta-Pei Road, Niao-Sung District, Kaohsiung City, 833, Taiwan. m93chinghua@gmail.com.
authors.forename: Ching-Hua
authors.initials: CH
authors.lastname: Hsieh
mesh_terms: D000818:Animals; D055354:Exosomes / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000378:metabolism*; D016328:NF-kappa B / Q000172:deficiency* / Q000235:genetics; D020543:Proteome; D015427:Reperfusion Injury / Q000503:physiopathology*; D015398:Signal Transduction
chemical_list: D016328:NF-kappa B; D020543:Proteome