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Aging and disease    2016, Vol. 7 Issue (5) : 623-634     DOI: 10.14336/AD.2016.0121
Original Article |
Proteomic Analysis of the Peri-Infarct Area after Human Umbilical Cord Mesenchymal Stem Cell Transplantation in Experimental Stroke
Dongsheng He1,2,3,5,6, Zhuo Zhang4, Jiamin Lao1,2,3,5,6, Hailan Meng1,2,3,5,6, Lijuan Han1,2,3,5,6, Fan Chen1,2,3,5,6, Dan Ye1,2,3,5,6, He Zhang1,2,3,5,6, Yun Xu1,2,3,5,6,*
1Department of Neurology, Affiliated Drum Tower Hospital, and
2Jiangsu Key Laboratory for Molecular Medicine, Nanjing University Medical School, Nanjing 210008, China.
3The State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, Nanjing 210008, China
4Department of Gastroenterology, Children’s Hospital of Nanjing, Nanjing Medical University, Nanjing 210008, China
5Jiangsu Province Stroke Center for Diagnosis and Therapy, Nanjing 210008, China
6Nanjing Neuropsychiatry Clinic Medical Center, Nanjing 210008, China
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Abstract  

Among various therapeutic approaches for stroke, treatment with human umbilical cord mesenchymal stem cells (hUC-MSCs) has acquired some promising results. However, the underlying mechanisms remain unclear. We analyzed the protein expression spectrum of the cortical peri-infarction region after ischemic stroke followed by treatment with hUC-MSCs, and found 16 proteins expressed differentially between groups treated with or without hUC-MSCs. These proteins were further determined by Gene Ontology term analysis and network with CD200-CD200R1, CCL21-CXCR3 and transcription factors. Three of them: Abca13, Grb2 and Ptgds were verified by qPCR and ELISA. We found the protein level of Abca13 and the mRNA level of Grb2 consistent with results from the proteomic analysis. Finally, the function of these proteins was described and the potential proteins that deserve to be further studied was also highlighted. Our data may provide possible underlying mechanisms for the treatment of stroke using hUC-MSCs.

Keywords stroke      hUC-MSCs      proteomics      iTRAQ      transplantation      human     
Corresponding Authors: Yun Xu   
About author:

These authors contributed equally to this work

Issue Date: 01 October 2016
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Dongsheng He
Zhuo Zhang
Jiamin Lao
Hailan Meng
Lijuan Han
Fan Chen
Dan Ye
He Zhang
Yun Xu
Cite this article:   
Dongsheng He,Zhuo Zhang,Jiamin Lao, et al. Proteomic Analysis of the Peri-Infarct Area after Human Umbilical Cord Mesenchymal Stem Cell Transplantation in Experimental Stroke[J]. Aging and disease, 2016, 7(5): 623-634.
URL:  
http://www.aginganddisease.org/EN/10.14336/AD.2016.0121     OR     http://www.aginganddisease.org/EN/Y2016/V7/I5/623
ExperimentGroupNo. of mice
Proteomicssham1-3
MCAO 24h4-6
MCAO 48h7-9
MCAO 48h + hUC-MSCs10-12

QPCR

Sham 6h

13-17
MCAO 6h18-22
MCAO + hUC-MSCs 6h23-27
Sham 12h28-32
MCAO 12h33-37
MCAO + hUC-MSCs 12h38-42
Sham 24h43-47
MCAO 24h48-52
MCAO + hUC-MSCs 24h53-57
Sham 48h58-62
MCAO 48h63-67
MCAO + hUC-MSCs 48h68-72
Sham 72h73-77
MCAO 72h78-82
MCAO + hUC-MSCs 72h83-87
Sham 1W88-92
MCAO 1W93-97
MCAO + hUC-MSCs 1W98-102

ELISA

Sham 24h

43-47
MCAO 24h48-52
MCAO + hUC-MSCs 24h53-57
Sham 48h58-62
MCAO 48h63-67
MCAO + hUC-MSCs 48h68-72
Sham 72h73-77
MCAO 72h78-82
MCAO + hUC-MSCs 72h83-87
Sham 1W88-92
MCAO 1W93-97
MCAO + hUC-MSCs 1W98-102
Table1  The number of mice in each experiment group.
mcao-24h/shammcao-48h/shammcao-48h/mcao-24hmcao-48h + hUC-MSCs/mcao-48h
Up-regulated proteinAbca13,Ptgds, Hbb-b1Abca13Acot11,Acat2,Grb2,Scp2,
Ppp2r5d,Rps12-ps11
Rrbp1, Anxa6, Slk

Down-regulated protein

Acot11,Acat2, Grb2,Scp2, Anp32b, Ppp2r5d

Acot11,Vps13d, Anp32b

Vps13d,Ptgds, Hbb-b1

Nup205, Psmd6
Table 2  Differentially expressed proteins in each group
Figure 1.  The number of overlapping proteins in the four groups. The blue circle represent the up-regulated and down-regulated proteins at 24 hours after cerebral ischemia compared to sham group, the yellow circle represent the up-regulated and down-regulated proteins at 48 hours after cerebral ischemia compared to sham group, the green circle represent up-regulated and down-regulated proteins at 48 hours after cerebral ischemia compared with the 24 hours after cerebral ischemia group, the red circle represent up-regulated and down-regulated proteins after hUC-MSCs treatment compared with the 48 hours after cerebral ischemia group.
Figure 2.  The GO term analysis of differentially expressed proteins in the four experimental groups. The GO term analysis of differentially expressed protein in the mcao-24h/sham group (A), the mcao-48h/sham group (B), the mcao-48h/mcao-24h group (C), and the mcao-48h + MSCs/mcao-48h group (D). Every GO term analysis shows genes involved in cellular component, molecular function and biological process.
Figure 3.  The network associated with CD200-CD200R1, CCL21-CXCR3 of differentially expressed proteins in the four experimental groups. The network with CD200-CD200R1 and CCL21-CXCR3 of differentially expressed proteins in the mcao-24h/sham group (A), the mcao-48h/sham group (B), the mcao-48h/mcao-24h group (C), and the mcao-48h + MSCs/mcao-48h group (D) are shown. Note that the trend in the mcao-24h/sham group was almost opposite to that of the mcao-48h/mcao-24h group.
Figure 4.  The network with transcription factors (c-myb, Runx-1, Pu.1, Irf8, Hoxb8) of differentially expressed proteins in the four experimental groups. The network with transcription factors (c-myb, Runx-1, Pu.1, Irf8, Hoxb8) of differential expression in the mcao-24h/sham group (A), the mcao-48h/sham group (B), the mcao-48h/mcao-24h group (C) and the mcao-48h + MSCs/mcao-48h group (D) are shown. The transcription factors of c-myb, Runx-1, Pu.1, Irf8, and Hoxb8 are represented by myb, runx1, Spi1, Irf8 and Hoxb8, respectively.
Figure 5.  The mRNA and protein levels of Abca13, Grb2, Ptgds in MCAO group and MCAO + MSCs group at indicated time points. The mRNA and protein levels of Abca13 are shown in (A) and (B), respectively. The mRNA and protein levels of Grb2 are shown in (C) and (D), respectively. The mRNA and protein levels of Ptgds are shown in (E) and (F), respectively. *p<0.05; **p<0.01 compared with sham group; #p<0.05; ##p<0.01 compared with MCAO group.
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