Quantitative analysis of protein and gene expression in salivary glands of Sjogren's-like disease NOD mice treated by bone marrow soup.
BACKGROUND
Bone marrow cell extract (termed as BM Soup) has been demonstrated to repair irradiated salivary glands (SGs) and restore saliva secretion in our previous study. In the present study, we aim to investigate if the function of damaged SGs in non-obese diabetic (NOD) mice can be restored by BM Soup treatment and the molecular alterations associated with the treatment.


METHODS
Whole BM cells were lysed and soluble intracellular contents ("BM Soup") were injected I.V. into NOD mice. Tandem mass tagging with 2-D liquid chromatography-mass spectrometry was used to quantify proteins in the submandibular glands (SMGs) between untreated and BM Soup-treated mice. Quantitative PCR was used to identify genes with altered expression in the treated mice.
restored salivary flow rates to normal levels and significantly reduced the focus scores of SMGs in NOD mice. More than 1800 proteins in SMG cells were quantified by the proteomic approach. Many SMG proteins involved in inflammation and apoptosis were found to be down-regulated whereas those involved in salivary gland biology and development/regeneration were up-regulated in the BM Soup-treated mice. qPCR analysis also revealed expression changes of growth factors and cytokines in the SMGs of the treated NOD mice.


CONCLUSIONS
BM Soup treatment is effective to restore the function of damaged SGs in NOD mice. Through gene/protein expression analysis, we have found that BM Soup treatment might effectuate via inhibiting apoptosis, focal adhesion and inflammation whereas promoting development, regeneration and differentiation of the SG cells in NOD mice. These findings provide important insights on the potential mechanisms underlying the BM Soup treatment for functional restoration of damaged SGs in NOD mice. Additional studies are needed to further confirm the identified target genes and their related signaling pathways that are responsible for the BM Soup treatment.
School of Dentistry, University of California Los Angeles, Los Angeles, California, United States of America.
Kaori
K
Misuno
Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, Montreal, Quebec, Canada.
Simon D
SD
Tran
Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, Montreal, Quebec, Canada.
Saeed
S
Khalili
School of Dentistry, University of California Los Angeles, Los Angeles, California, United States of America.
Junwei
J
Huang
Craniofacial Tissue Engineering and Stem Cells Laboratory, Faculty of Dentistry, McGill University, Montreal, Quebec, Canada.
Younan
Y
Liu
School of Dentistry, University of California Los Angeles, Los Angeles, California, United States of America.
Shen
S
Hu
D000818:Animals; D001854:Bone Marrow Cells / Q000737:chemistry; D002457:Cell Extracts / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D008297:Male; D016688:Mice, Inbred NOD; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D012859:Sjogren's Syndrome / Q000188:drug therapy / Q000378:metabolism*; D013363:Submandibular Gland / Q000187:drug effects / Q000378:metabolism*; D059467:Transcriptome
D002457:Cell Extracts; D020543:Proteome