Urinary Proteomic Study On Mucopolysaccharidosis Type ?

Objective:Study on urinary differential proteomics of mucopolysaccharidosis type ?(MPS ?)1.To find urine biomarkers of MPS ? for disease diagnostic and prognosis monitoring.2.To provide basis for illuminating the pathphysiological mechanisms of MPS ?.Methods:From March 2015 to June 2016,12 patients with MPS ?(MPS ? group)and 15 healthy people(healthy control group)were enrolled and collected morning urine samples.1.Two dimensional gel electrophoresis(2-DE)were used to separate total protein of MPS ? group and healthy control group respectively,followed by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS)to identify the protein.2.Urinary a 1-antitrypsin(?1-AT),Gm2 activator protein(GM2A),vitamin D-binding protein(DBP)and lipocalin-type prostaglandin D synthase(L-PGDS)of each groups were detected using the methods of ELISA and Western Blot.3.After protein preparation,the peptides were labeled with isobaric tags for relative and absolute quantification(iTRAQ).After that protein were identified with liquid chromatography-mass spectrometer/mass spectrometer,(LC-MS/MS).Bioinformatics analysis for differential expressed proteins including Molecular Function,Cellular Component and Biological Process,then the Gene Ontology(GO)annotation,protein-protein interaction analysis and enrichment analysis of pathway were conducted to determine the metabolic pathway and the signal transduction pathways which differential proteins involved.Results:1.Comparing the MPS ? group and healthy control group,32 differential protein spots were found by using 2-DE,and 20 differential proteins were identified by MALDI-TOF-MS.The number of up-regulated in MPS ? group is 10,and on the contrary are 10.2.The value of urinary ?1-AT/Cr and DBP/Cr in MPS ? group were 55.89±7.43 mg/mmol and 1.31 ±0.14 ng/mmol,which were significantly higher than 33.32±3.59 mg/mmol and 0.14±0.02 mg/mmol in healthy control group(P<0.05);while the value of urinary GM2A/Cr and L-PGDS/Cr in MPS ? group were 2.68 ±0.44 ng/mmol and 8.02 ±2.04 pg/mmol,which were significantly lower than 0.35±0.06 ng/mmol and 18.14± 1.67 pg/mmol in healthy control group(P<0.05).3 Comparing the MPS II group and healthy control group,693 differential proteins were identifies by using iTRAQ and LC-MS/MS.Many of the proteins are involved in spliceosome,ribosome,stress response,oxidative phosphorylation and complement system.Conclusion:1.Compare to the healthy control group,urinary al-AT/Cr,DBP/Cr were found upregulated in MPS ?,while urinary GM2A/Cr and L-PGDS/Cr were found downregulated,which can be considered as accessory diagnostic biomarkers for MPS?.2.Pathology changes of MPS ? may be associated with abnormity in signaling pathway of spliceosome,ribosome,stress response,oxidative phosphorylation and complement system,and the results pave the way to clarify pathogenic mechanism of MPS ?.