To be able to discriminate proteomic changes implicated in either IR-induced digestive tract damage or rescuing through hPDSC administration, we carried out comparative proteomic analysis using label-free quantification analysis along with quadrupole time-of-flight (Q-TOF) mass spectrometry. As results, because seen in Fig.   5a , all of us pulled out whole proteomes from group 1 ( n   =  186), team 2 ( n   =  270), and group 3 ( n   =  238) and performed a comparative gene ontology (GO) analysis. These 143 spots, including three groups in common were changed based on group condition (see Additional file 1 : Table S3). Upon the serial comparison analysis, we categorized these proteome spots into 5 groups (Fig.   5b ). Briefly, we could confirm that hPDSCs imposed anti-oxidative, cell difference, and tissue repair, whereas they resulted in the inhibited of signal transduction relevant to inflammation, oxidative stress, plus ion transport and the changing patterns could be categorized in to five patterns as seen in Fig.   5b , among which the following three pattern forms were prominent, that is, proteome spots significantly decreased within group 2, whereas significantly increased in group a few (Fig.   5c plus Table  1 ), proteomic areas significantly increased in group 2, but significantly reduced in group 3 (Fig.   5d and Table  1 ), and proteomic spots were not changed in team 2, but significantly increased in group 3 (Fig.   5e and Table  1 ). The proteomic places identified as proteomic biomarkers signifying “ elevated following IR, but decreased following administration of hPDSCs” were phosphatidylethanolamine-binding protein 1 (PEBP1), apoptosis-associated speck-like protein containing CREDIT CARD (PYCARD), glycerol 3-phosphate dehydrogenase [NAD (+)], cytoplasmic (GPD1) and ornithine carbamoyltransferase, mitochondria (OTC) (Fig.   5d and Table  1 ), and three proteomic spots, recognized as proteomic biomarkers containing “ no significant changes subsequent IR, but increased following administration of hPDSCs” had been tubulin-α -1B chain (TUBA1B), cytochrome c oxidase subunit 2 (MT-CO2), and peroxiredoxin-2 (PRDX-2) (Fig  5e and Table  1 ). To draw significant proteomic biomarkers with validation, proteomic biomarkers signifying “ downregulated following IR, but improved following administration of hPDSCs”, IL-10, TIMP-1, and GST mu kind 1 (GST mu 1) were done with Western blot using homogenates through each group (Fig.   6a and b ). As validation to these discoveries, as shown in Fig.   6a , the expression degrees of IL-10 were decreased with irradiation ( P L 21 ]. TIMP-1 is a glycoprotein expressed from several tissues [ 22 ] as a natural inhibitor of MMPs [ 23 ], and has also been reported to reflect protection against IR-induced tissue damage. As shown in Fig.   6a , TIMP-1 was decreased in group two ( P P 3c , MMP-2 activity was significantly increased in team 2 . GST mu type— known as a strong anti-oxidant marker— was upregulated in group 3 whereas a decrease in expression of the protein was observed in group 2 ( P 6b ). Cancellation of NOX-4, OTC and GPD1 has been validated as a biomarker predicting favorable response with hPDSC administration (Fig.   6c ). As a biomarker unchanged as a result of IR treatment, but improved following hPDSC administration, GST (pi) and PRDX-2— termed as a strongly anti-oxidative factor— was validated ( P 6d ).

Fig. 5

Label-free quantification analysis to detect biomarkers. the Venn diagram of identified proteins based on groups. b Gene ontology categories and the number of genes with expression variations are depicted in the boxes . c e Novel biomarkers of each group subsequent Table  1 . GPD1 g lycerol 3-phosphate dehydrogenase [NAD (+)], cytoplasmic, GST mu one , glutatione-S-transferase mu 1, MT-CO2 cytochrome c oxidase subunit 2 OTC o rnithine carbamoyltransferase, mitochondria PEBP1 phosphatidylethanolamine-binding protein one; PRDX-2 peroxiredoxin-2, PYCARD apoptosis-associated speck-like proteins containing CARD, TUBA1B tubulin-α -1B chain; MPTX mucosal pentraxin protein

Table 1

Label-free quantification analysis: novel biomarker applicants

Proteomes elevated with irradiation (G2) when compared with control (G1), but decreased with hPDSC (G3)

Protein description

G1

G2

G3

Fatty acid-binding protein, adipocyte

1

1 . 6125

0. 7874

Heat shock protein 75  kDa, mitochondrial

1

1 . 7864

0. 9751

RIKEN cDNA 2210010C04, isoform CRA_b

1

2 . 9038

1 . 1034

Neurobeachin-like protein 2

1

7. 2087

3. 2829

Gelsolin

1

9. 6935

3. 3595

Nucleolin

one

eleven. 1004

5. 0287

Phosphatidylethanolamine-binding proteins 1

1

12. 4758

1 . 8796

Protein 6530409C15Rik

1

15. 1723

nine. 6556

Apoptosis-associated speck-like proteins containing a CARD

1

24. 5311

0. 9818

Nucleoside diphosphate kinase B

1

25. 2488

15. 8437

Glycerol-3-phosphate dehydrogenase [NAD(+)], cytoplasmic

1

38. 2966

21. 2440

Tropomyosin alpha-3 chain

1

40. 7211

0. 30890

60S ribosomal protein L22

1

598. 3902

127. 068

Ornithine carbamoyltransferase, mitochondrial

1

249659

163892

Histone H4

1

609717

304923

Proteomes decreased after irradiation (G2) compared to control (G1), but increased with hPDSC (G3)

Proteins Mptx 2

1

0. 1956

0. 5902

Glutathione S-transferase Mu 1

1

0. 2547

7. 3523

Proteomes not changed along with irradiation (G2) compared to control (G1), but increased along with hPDSC (G3)

Profilin-1

1

0. 7039

3. 2086

Myosin-10

1

0. 8149

11. 362

Tubulin alpha-1B chain

1

0. 8376

17. 0778

Macoilin

1

0. 8532

6. 89827

Cytochrome c oxidase subunit 2

1

0. 9668

2 . 58191

Gastrotropin

1

1 . 09828

1 . 81219

Peptidyl-prolyl cis-trans isomerase The

one

1 ) 3439

21. 883

Maltase-glucoamylase

1

1 . 3488

6. 0850

Peroxiredoxin-2

1

1 . 4326

4. 2072

Histone H2A type 1-F

1

1 . 4423

4. 6122

Calnexin

1

1 . 4684

second . 8897

Fig. 6

Validation of identified target through proteomics analysis. a The expressions of IL-10 and TIMP1 were researched by RT-PCR. b The GST ( mu ) levels were determined by Western blotting on experimental organizations. c The NOX4, OTC, and GPD levels were determined by Western blotting on experimental groups. d The GST (pi) and PRDX2 levels had been determined by Western blotting on experimental groups. GPD1 g lycerol 3-phosphate dehydrogenase [NAD (+)], cytoplasmic, GST mu one , glutatione-S-transferase mu 1, IL-6 interleukin 6, NOX-4 NADPH-oxidase 4, OVER THE COUNTER ornithine carbamoyltransferase, mitochondria, PRDX-2 peroxiredoxin-2, TIMP-1 tissue inhibitor of metalloproteinases