Proteomic analysis of tumor necrosis factor-alpha-induced secretome of human adipose tissue-derived mesenchymal stem cells.
Human adipose tissue-derived mesenchymal stem cells (hASCs) are useful for regeneration of inflamed or injured tissues. To identify secreted hASC proteins during inflammation, hASCs were exposed to tumor necrosis factor-alpha (TNF-alpha) and conditioned media derived from hASCs were analyzed by liquid chromatography coupled with tandem mass spectrometry. We identified 187 individual proteins as secreted proteins (secretome) in hASC-conditioned media; 118 proteins were secreted at higher levels upon TNF-alpha treatment. The TNF-alpha-induced secretome included a variety of cytokines and chemokines such as interleukin-6 (IL-6), IL-8, chemokine (C-X-C motif) ligand 6, and monocyte chemotactic protein-1 (MCP-1). TNF-alpha also increased expression of various proteases including cathepsin L, matrix metalloproteases and protease inhibitors, and induced secretion of long pentraxin 3, a key inflammatory mediator implicated in innate immunity. TNF-alpha-conditioned media stimulated migration of human monocytes, which play a key role in inflammatory responses. This migration was abrogated by pretreatment with neutralizing anti-IL-6, anti-IL-8, and anti-MCP-1 antibodies, suggesting that IL-6, IL-8, and MCP-1 are involved in migration of monocytes. Taken together, these results suggest that TNF-alpha-induced secretome may play a pivotal role in inflammatory responses and that shotgun proteomic analysis will be useful for elucidation of the paracrine functions of mesenchymal stem cells.
Lee
Mi Jeong
MJ
Medical Research Center for Ischemic Tissue Regeneration & Medical Research Institute, College of Medicine, Pusan National University, Yangsan, Republic of Korea.
Kim
Jaeyoon
J
Kim
Min Young
MY
Bae
Yoe-Sik
YS
Ryu
Sung Ho
SH
Lee
Taehoon G
TG
Kim
Jae Ho
JH
D000273:Adipose Tissue / Q000166:cytology*; D000704:Analysis of Variance; D002403:Cathepsins / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology; D002853:Chromatography, Liquid; D016207:Cytokines / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism; D020543:Proteome / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000494:pharmacology*
D016207:Cytokines; D020543:Proteome; D014409:Tumor Necrosis Factor-alpha; D002403:Cathepsins; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1