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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

The morphological changes of cultured endothelial progenitor cells (EPCs). All images were taken under light microscope. Day-1, cells were small-round-shaped; Day-3, cells enlarged; Day-7, cells were short spindle sharped; Day-14, cells became slender-spindle-shaped and number of cells increased; Day-21, cells grew with a “cobblestone” appearance.
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Figure 2: The morphological changes of cultured endothelial progenitor cells (EPCs). All images were taken under light microscope. Day-1, cells were small-round-shaped; Day-3, cells enlarged; Day-7, cells were short spindle sharped; Day-14, cells became slender-spindle-shaped and number of cells increased; Day-21, cells grew with a “cobblestone” appearance.

Mentions: The freshly isolated EPCs were small-round shaped and suspended in culture medium. At day 2, cells started to attach to the dish wall, and cellular hypertrophy was observed at day 3, followed by becoming short spindle shaped. The cellular growth peak was reached after 7-day culture. At day 10–14, the cells became slender-spindle-shaped and started colony formation. At day 21, cell colony formation was completed with a “cobblestone” appearance, as shown in Figure 2. At day 14, EPCs were identified by CD34, CD133, FLK-1 using flow cytometer. DiL-acLDL (red)/FITC-UEA-1 (green) dual staining cells were observed under the laser scanning confocal microscopy. The cells with dual staining (shown as yellow fluorescence) were considered as differentiating EPCs (Figure 3).


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

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

The morphological changes of cultured endothelial progenitor cells (EPCs). All images were taken under light microscope. Day-1, cells were small-round-shaped; Day-3, cells enlarged; Day-7, cells were short spindle sharped; Day-14, cells became slender-spindle-shaped and number of cells increased; Day-21, cells grew with a “cobblestone” appearance.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 2: The morphological changes of cultured endothelial progenitor cells (EPCs). All images were taken under light microscope. Day-1, cells were small-round-shaped; Day-3, cells enlarged; Day-7, cells were short spindle sharped; Day-14, cells became slender-spindle-shaped and number of cells increased; Day-21, cells grew with a “cobblestone” appearance.
Mentions: The freshly isolated EPCs were small-round shaped and suspended in culture medium. At day 2, cells started to attach to the dish wall, and cellular hypertrophy was observed at day 3, followed by becoming short spindle shaped. The cellular growth peak was reached after 7-day culture. At day 10–14, the cells became slender-spindle-shaped and started colony formation. At day 21, cell colony formation was completed with a “cobblestone” appearance, as shown in Figure 2. At day 14, EPCs were identified by CD34, CD133, FLK-1 using flow cytometer. DiL-acLDL (red)/FITC-UEA-1 (green) dual staining cells were observed under the laser scanning confocal microscopy. The cells with dual staining (shown as yellow fluorescence) were considered as differentiating EPCs (Figure 3).

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