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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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mRNA level of MCP-1, CD105 and HIF-1α. Real-time quantitative RT-PCR analysis of MCP-1, CD105 and HIF-1a mRNA after transplantation. **P < 0. 05 vs normal, *P < 0.05 vs IgAN. (A). MCP-1, CD105 and HIF-1a mRNA from the renal cortex of rats isolated from normal, day-0, day −1, day-3, day-7 and day14 EPCs rats after transplantation. *P < 0.05,**P < 0.01, comparing to the day-0 group (B, C, D).
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Figure 9: mRNA level of MCP-1, CD105 and HIF-1α. Real-time quantitative RT-PCR analysis of MCP-1, CD105 and HIF-1a mRNA after transplantation. **P < 0. 05 vs normal, *P < 0.05 vs IgAN. (A). MCP-1, CD105 and HIF-1a mRNA from the renal cortex of rats isolated from normal, day-0, day −1, day-3, day-7 and day14 EPCs rats after transplantation. *P < 0.05,**P < 0.01, comparing to the day-0 group (B, C, D).

Mentions: To confirm the expression prolife of the molecules we observed in immuno- histochemistry studies, we measured the expression again by using quantitative western blotting or quantitative RT-PCR. Similarly, we found that normal group express relatively less amount of MCP-1, HIF-1α, which were significantly higher in IgAN groups. However, after EPC transplantation, MCP-1, HIF-1α expression was gradually decreased in 14 days. The protein expression of CD105 was decreased in the first 3 days after transplantation, while it increased in day-7 and day-14. mRNA of these molecules showed the similar results. mRNA levels of MCP-1 and HIF-1α were significantly increased in IgAN group, while the levels were gradually decreased in 14 days after transplantation. CD105’s mRNA first increased in the first 7 days after transplantation, but it increased and reached the peak at day-14 (Figures 7, 8 & 9).


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

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

mRNA level of MCP-1, CD105 and HIF-1α. Real-time quantitative RT-PCR analysis of MCP-1, CD105 and HIF-1a mRNA after transplantation. **P < 0. 05 vs normal, *P < 0.05 vs IgAN. (A). MCP-1, CD105 and HIF-1a mRNA from the renal cortex of rats isolated from normal, day-0, day −1, day-3, day-7 and day14 EPCs rats after transplantation. *P < 0.05,**P < 0.01, comparing to the day-0 group (B, C, D).
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 9: mRNA level of MCP-1, CD105 and HIF-1α. Real-time quantitative RT-PCR analysis of MCP-1, CD105 and HIF-1a mRNA after transplantation. **P < 0. 05 vs normal, *P < 0.05 vs IgAN. (A). MCP-1, CD105 and HIF-1a mRNA from the renal cortex of rats isolated from normal, day-0, day −1, day-3, day-7 and day14 EPCs rats after transplantation. *P < 0.05,**P < 0.01, comparing to the day-0 group (B, C, D).
Mentions: To confirm the expression prolife of the molecules we observed in immuno- histochemistry studies, we measured the expression again by using quantitative western blotting or quantitative RT-PCR. Similarly, we found that normal group express relatively less amount of MCP-1, HIF-1α, which were significantly higher in IgAN groups. However, after EPC transplantation, MCP-1, HIF-1α expression was gradually decreased in 14 days. The protein expression of CD105 was decreased in the first 3 days after transplantation, while it increased in day-7 and day-14. mRNA of these molecules showed the similar results. mRNA levels of MCP-1 and HIF-1α were significantly increased in IgAN group, while the levels were gradually decreased in 14 days after transplantation. CD105’s mRNA first increased in the first 7 days after transplantation, but it increased and reached the peak at day-14 (Figures 7, 8 & 9).

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