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Transplantation of endothelial progenitor cells in treating rats with IgA nephropathy.

Guo W, Feng JM, Yao L, Sun L, Zhu GQ - BMC Nephrol (2014)

Bottom Line: The transplanted BM-EPCs were successfully located in IgAN rat kidney.After transplantation, Urinary red blood cell, urine protein, BUN, Scr and IgA serum level were significantly decreased, so were the areas of glomerular extracellular matrix and the IgA deposition in the glomeruli.And the expression levels of HIF-1α and MCP-1 were significantly down-regulated, while the expression of CD105 was up-regulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Nephrology, The First Affiliated Hospital of China Medical University, Shenyang 110001, China. fengjiangm@163.com.

ABSTRACT

Background: Therapeutic options in IgAN are still limited. The aim of this study is to explore the feasibility of using endothelial progenitor cell to treat IgAN in rat model.

Methods: Rat bone marrow mononuclear cells (BM-MNCs) obtained with density gradient centrifugation were cultured in vitro, and induced into endothelial progenitor cells (EPCs). EPCs were identified by surface marker CD34, CD133 and VEGFR2 (FLK-1) and by Dil-Ac-LDL/FITC-UEA-1 double staining. EPCs were labeled with PKH26 prior to transplantation. Rat model of IgAN was established by oral administration of bovine serum albumin together with lipopolysaccharide via the caudal vein and subcutaneous injection of CCL4. Kidney paraffin sections were stained by H&E and PAS. Immunofluorescence was used to assess IgA deposition in the glomeruli. Peritubular capillary (PTC) density was determined by CD31 staining. Monocyte chemoattrant protein-1 (MCP-1), hypoxia-inducible factor-1α (HIF-1α) and CD105 were also measured by immunohistochemistry, western blotting and real-time fluorescent quantitative PCR.

Results: The transplanted BM-EPCs were successfully located in IgAN rat kidney. After transplantation, Urinary red blood cell, urine protein, BUN, Scr and IgA serum level were significantly decreased, so were the areas of glomerular extracellular matrix and the IgA deposition in the glomeruli. In addition, PTC density was elevated. And the expression levels of HIF-1α and MCP-1 were significantly down-regulated, while the expression of CD105 was up-regulated. All these changes were not observed in control groups.

Conclusion: The BM-EPCs transplantation significantly decreases the expansion of glomerular extracellular matrix and the deposition of IgA in the glomeruli; lowers the expression of inflammatory factors; increases PTC density; improves ischemic-induced renal tissue hypoxia, all of which improves the renal function and slows the progress of IgA nephropathy.

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Related in: MedlinePlus

Immunostaining of PKH26 on tissues from IgAN rats after EPCs transplantation. In kidney tissue, the white arrows indicate the location of glomeruli. The intensity of PKH26 positive signal were quantified With MetaMorph software, and presented in column graph (bottom right). *P < 0.05, **P < 0.01, comparing to pre- transplantation, #P < 0.05, comparing today-1 (A). PKH26-positive signal in the liver tissue (B).
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Figure 4: Immunostaining of PKH26 on tissues from IgAN rats after EPCs transplantation. In kidney tissue, the white arrows indicate the location of glomeruli. The intensity of PKH26 positive signal were quantified With MetaMorph software, and presented in column graph (bottom right). *P < 0.05, **P < 0.01, comparing to pre- transplantation, #P < 0.05, comparing today-1 (A). PKH26-positive signal in the liver tissue (B).

Mentions: Cryo-section of kidney and liver tissues stained with PKH26 and incubated with 4′, 6-diamidino-2-phenylindole (DAPI). PKH26 positive cells showed red fluorescence, nuclei were stained blue with DAPI under fluorescence microscopy. 1 day after transplantation, PKH26 positive cells were detected in the kidney of EPCs rats, mainly in the interstitial area. 3, 7, 14 days after transplantation, more and more PKH26 positive cells were discovered in glomerular and interstitial areas. The intensity of red fluorescence was gradually increased after transplantation. The difference between day-1 and day-14 after transplantation was significant. In addition to the kidney, PKH26-positive signal could also be detected in the liver tissue. The significance of this liver migration of EPCs was still waited to be investigated (Figure 4).


Transplantation of endothelial progenitor cells in treating rats with IgA nephropathy.

Guo W, Feng JM, Yao L, Sun L, Zhu GQ - BMC Nephrol (2014)

Immunostaining of PKH26 on tissues from IgAN rats after EPCs transplantation. In kidney tissue, the white arrows indicate the location of glomeruli. The intensity of PKH26 positive signal were quantified With MetaMorph software, and presented in column graph (bottom right). *P < 0.05, **P < 0.01, comparing to pre- transplantation, #P < 0.05, comparing today-1 (A). PKH26-positive signal in the liver tissue (B).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4109798&req=5

Figure 4: Immunostaining of PKH26 on tissues from IgAN rats after EPCs transplantation. In kidney tissue, the white arrows indicate the location of glomeruli. The intensity of PKH26 positive signal were quantified With MetaMorph software, and presented in column graph (bottom right). *P < 0.05, **P < 0.01, comparing to pre- transplantation, #P < 0.05, comparing today-1 (A). PKH26-positive signal in the liver tissue (B).
Mentions: Cryo-section of kidney and liver tissues stained with PKH26 and incubated with 4′, 6-diamidino-2-phenylindole (DAPI). PKH26 positive cells showed red fluorescence, nuclei were stained blue with DAPI under fluorescence microscopy. 1 day after transplantation, PKH26 positive cells were detected in the kidney of EPCs rats, mainly in the interstitial area. 3, 7, 14 days after transplantation, more and more PKH26 positive cells were discovered in glomerular and interstitial areas. The intensity of red fluorescence was gradually increased after transplantation. The difference between day-1 and day-14 after transplantation was significant. In addition to the kidney, PKH26-positive signal could also be detected in the liver tissue. The significance of this liver migration of EPCs was still waited to be investigated (Figure 4).

Bottom Line: The transplanted BM-EPCs were successfully located in IgAN rat kidney.After transplantation, Urinary red blood cell, urine protein, BUN, Scr and IgA serum level were significantly decreased, so were the areas of glomerular extracellular matrix and the IgA deposition in the glomeruli.And the expression levels of HIF-1α and MCP-1 were significantly down-regulated, while the expression of CD105 was up-regulated.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Nephrology, The First Affiliated Hospital of China Medical University, Shenyang 110001, China. fengjiangm@163.com.

ABSTRACT

Background: Therapeutic options in IgAN are still limited. The aim of this study is to explore the feasibility of using endothelial progenitor cell to treat IgAN in rat model.

Methods: Rat bone marrow mononuclear cells (BM-MNCs) obtained with density gradient centrifugation were cultured in vitro, and induced into endothelial progenitor cells (EPCs). EPCs were identified by surface marker CD34, CD133 and VEGFR2 (FLK-1) and by Dil-Ac-LDL/FITC-UEA-1 double staining. EPCs were labeled with PKH26 prior to transplantation. Rat model of IgAN was established by oral administration of bovine serum albumin together with lipopolysaccharide via the caudal vein and subcutaneous injection of CCL4. Kidney paraffin sections were stained by H&E and PAS. Immunofluorescence was used to assess IgA deposition in the glomeruli. Peritubular capillary (PTC) density was determined by CD31 staining. Monocyte chemoattrant protein-1 (MCP-1), hypoxia-inducible factor-1α (HIF-1α) and CD105 were also measured by immunohistochemistry, western blotting and real-time fluorescent quantitative PCR.

Results: The transplanted BM-EPCs were successfully located in IgAN rat kidney. After transplantation, Urinary red blood cell, urine protein, BUN, Scr and IgA serum level were significantly decreased, so were the areas of glomerular extracellular matrix and the IgA deposition in the glomeruli. In addition, PTC density was elevated. And the expression levels of HIF-1α and MCP-1 were significantly down-regulated, while the expression of CD105 was up-regulated. All these changes were not observed in control groups.

Conclusion: The BM-EPCs transplantation significantly decreases the expansion of glomerular extracellular matrix and the deposition of IgA in the glomeruli; lowers the expression of inflammatory factors; increases PTC density; improves ischemic-induced renal tissue hypoxia, all of which improves the renal function and slows the progress of IgA nephropathy.

Show MeSH
Related in: MedlinePlus