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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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Immunohistochemical detection of CD31 , MCP-1, CD105 and HIF-1a. Normal group, CD31-positive endothelial cells were regularly distributed in renal interstitium (A); The expression of CD31 was significantly decreased in IgAN group (B); In EPCs group, the expression of CD31 was also decreased than the normal group,however, it was significantly increased compared to the one in IgAN group. (C). In normal group, MCP-1 staining was barely detectable in renal tubular epithelial cells (D); However, in IgAN group, the enhanced expression of MCP-1 was detected. (E); MCP-1 expression gradually decreased in EPCs group (F). Normal group, CD105 staining was weak. (G); IgAN group, the expression of CD105 was significantly elevated. (H); CD105 expression in EPCs group after transplantation. (I). In normal group, minimal HIF-1α was detected in renal tubular epithelial. (J); The expression of HIF-1α increased significantly in IgAN group, (K); It gradually decreased in EPCs group (L). Immunohistochemistry of CD31 (A, B, C); Immunohistochemistry of MCP-1 (D, E, F); Immunohistochemistry of CD105 (G, H, I); Immunohistochemistry of HIF-1α (J, K, L).
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Figure 6: Immunohistochemical detection of CD31 , MCP-1, CD105 and HIF-1a. Normal group, CD31-positive endothelial cells were regularly distributed in renal interstitium (A); The expression of CD31 was significantly decreased in IgAN group (B); In EPCs group, the expression of CD31 was also decreased than the normal group,however, it was significantly increased compared to the one in IgAN group. (C). In normal group, MCP-1 staining was barely detectable in renal tubular epithelial cells (D); However, in IgAN group, the enhanced expression of MCP-1 was detected. (E); MCP-1 expression gradually decreased in EPCs group (F). Normal group, CD105 staining was weak. (G); IgAN group, the expression of CD105 was significantly elevated. (H); CD105 expression in EPCs group after transplantation. (I). In normal group, minimal HIF-1α was detected in renal tubular epithelial. (J); The expression of HIF-1α increased significantly in IgAN group, (K); It gradually decreased in EPCs group (L). Immunohistochemistry of CD31 (A, B, C); Immunohistochemistry of MCP-1 (D, E, F); Immunohistochemistry of CD105 (G, H, I); Immunohistochemistry of HIF-1α (J, K, L).

Mentions: In normal group, CD31 was shown as brown fine deposits on the membrane of endothelial cells of PTC, which were distributed in renal interstitium among renal tubules; some was shown on the membrane of glomerular vascular endothelial cells. The expression of CD31 was significantly decreased in IgAN group as a comparison to the normal group, while the expression of CD31 in EPCs group was significantly increased compared to the one in IgAN group. Integrated optical density (IOD) value of CD31 was used to represent the density of peritubular capillary. The difference between groups was statistically significant (Figure 6, Tables 5 & 6).In normal group, MCP-1 staining was barely detectable in renal tubular epithelial cells. However, in IgAN group, the enhanced expression of MCP-1 was detected. After EPCs transplantation, MCP-1 expression gradually decreased at day-3, day-7 and day-14 (Figure 6).In normal group, CD105 staining was weak in stromal vascular endothelial cells and glomerular/interstitial cells. While in IgAN group, the expression of CD105 was significantly elevated prior to transplantation. CD105 expression in EPCs group diminished at day-3 after EPCs transplantation, while it increased at day 7 and day 14 (Figure 6).In normal group, minimal HIF-1α was detected in renal tubular epithelial. The expression of HIF-1α increased significantly in IgAN group, while it gradually decreased in EPCs group (Figure 6).


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

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

Immunohistochemical detection of CD31 , MCP-1, CD105 and HIF-1a. Normal group, CD31-positive endothelial cells were regularly distributed in renal interstitium (A); The expression of CD31 was significantly decreased in IgAN group (B); In EPCs group, the expression of CD31 was also decreased than the normal group,however, it was significantly increased compared to the one in IgAN group. (C). In normal group, MCP-1 staining was barely detectable in renal tubular epithelial cells (D); However, in IgAN group, the enhanced expression of MCP-1 was detected. (E); MCP-1 expression gradually decreased in EPCs group (F). Normal group, CD105 staining was weak. (G); IgAN group, the expression of CD105 was significantly elevated. (H); CD105 expression in EPCs group after transplantation. (I). In normal group, minimal HIF-1α was detected in renal tubular epithelial. (J); The expression of HIF-1α increased significantly in IgAN group, (K); It gradually decreased in EPCs group (L). Immunohistochemistry of CD31 (A, B, C); Immunohistochemistry of MCP-1 (D, E, F); Immunohistochemistry of CD105 (G, H, I); Immunohistochemistry of HIF-1α (J, K, L).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Figure 6: Immunohistochemical detection of CD31 , MCP-1, CD105 and HIF-1a. Normal group, CD31-positive endothelial cells were regularly distributed in renal interstitium (A); The expression of CD31 was significantly decreased in IgAN group (B); In EPCs group, the expression of CD31 was also decreased than the normal group,however, it was significantly increased compared to the one in IgAN group. (C). In normal group, MCP-1 staining was barely detectable in renal tubular epithelial cells (D); However, in IgAN group, the enhanced expression of MCP-1 was detected. (E); MCP-1 expression gradually decreased in EPCs group (F). Normal group, CD105 staining was weak. (G); IgAN group, the expression of CD105 was significantly elevated. (H); CD105 expression in EPCs group after transplantation. (I). In normal group, minimal HIF-1α was detected in renal tubular epithelial. (J); The expression of HIF-1α increased significantly in IgAN group, (K); It gradually decreased in EPCs group (L). Immunohistochemistry of CD31 (A, B, C); Immunohistochemistry of MCP-1 (D, E, F); Immunohistochemistry of CD105 (G, H, I); Immunohistochemistry of HIF-1α (J, K, L).
Mentions: In normal group, CD31 was shown as brown fine deposits on the membrane of endothelial cells of PTC, which were distributed in renal interstitium among renal tubules; some was shown on the membrane of glomerular vascular endothelial cells. The expression of CD31 was significantly decreased in IgAN group as a comparison to the normal group, while the expression of CD31 in EPCs group was significantly increased compared to the one in IgAN group. Integrated optical density (IOD) value of CD31 was used to represent the density of peritubular capillary. The difference between groups was statistically significant (Figure 6, Tables 5 & 6).In normal group, MCP-1 staining was barely detectable in renal tubular epithelial cells. However, in IgAN group, the enhanced expression of MCP-1 was detected. After EPCs transplantation, MCP-1 expression gradually decreased at day-3, day-7 and day-14 (Figure 6).In normal group, CD105 staining was weak in stromal vascular endothelial cells and glomerular/interstitial cells. While in IgAN group, the expression of CD105 was significantly elevated prior to transplantation. CD105 expression in EPCs group diminished at day-3 after EPCs transplantation, while it increased at day 7 and day 14 (Figure 6).In normal group, minimal HIF-1α was detected in renal tubular epithelial. The expression of HIF-1α increased significantly in IgAN group, while it gradually decreased in EPCs group (Figure 6).

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