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Updated: Jun 28, 2026
Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction
Published on: August 27, 2020
Endostatin, an antiangiogenic protein, is expressed in the unilateral ureteral obstruction mice model
Thiago T Maciel1, Enia L Coutinho, Debora Soares
1Nephrology Division, Federal University of São Paulo, São Paulo - Brazil. thiagotrovati@uol.com.br
Background:
Extracellular matrix accumulation, epithelial-to-mesenchymal transition, tubular atrophy and loss of peritubular capillary network are hallmarks of tubulointerstitial injury in progressive renal diseases. In this study, we analyzed endostatin expression in kidneys subjected to unilateral ureteral obstruction (UUO).
Methods:
Collagen XVIII mRNA expression was evaluated by real-time polymerase chain reaction (PCR). Endostatin and CD31 protein levels were analyzed by Western blot and immunohistochemistry. In vitro quantification of collagen XVIII and fibrosis-related genes in HK2 cells was performed by real-time PCR.
Results:
UUO significantly increased collagen XVIII mRNA expression and released a 30-kDa endostatin fragment. Immunohistochemistry revealed endostatin expression increased in injured tissue, mainly on tubular cells. Of interest, expression of CD31 was significantly reduced by UUO. Endostatin administration in vitro did not modify the expression of genes related to fibrosis development. However, in vitro TGF-beta1 administration induced expression of collagen XVIII/endostatin mRNA in human tubular cells.
Conclusion:
Endostatin is expressed during the progression of renal fibrosis in vitro and in vivo, suggesting a role for endostatin in development of tubulointerstitial injury.
Insights
Endostatin expression increases during kidney fibrosis development, particularly in tubular cells, following unilateral ureteral obstruction (UUO). This suggests endostatin plays a role in progressive renal tubulointerstitial injury.
Area of Science:
- Nephrology
- Molecular Biology
- Pathology
Background:
- Tubulointerstitial injury is characterized by extracellular matrix accumulation, epithelial-to-mesenchymal transition, tubular atrophy, and reduced peritubular capillaries.
- Progressive renal diseases involve complex cellular and molecular changes in the kidney's interstitium.
Purpose of the Study:
- To investigate the expression and role of endostatin in the context of kidney injury induced by unilateral ureteral obstruction (UUO).
- To determine the relationship between endostatin, collagen XVIII, and fibrosis markers in renal disease models.
Main Methods:
- Real-time polymerase chain reaction (PCR) to quantify Collagen XVIII mRNA.
- Western blot and immunohistochemistry to analyze endostatin and CD31 protein levels.
- In vitro studies using HK2 cells to assess gene expression changes.
Main Results:
- Unilateral ureteral obstruction (UUO) significantly upregulated Collagen XVIII mRNA and released a 30-kDa endostatin fragment.
- Endostatin expression was elevated in injured kidney tissue, primarily localized to tubular cells, while CD31 expression decreased.
- In vitro, TGF-beta1 induced Collagen XVIII/endostatin mRNA in human tubular cells, but endostatin administration alone did not affect fibrosis gene expression.
Conclusions:
- Endostatin is demonstrably expressed during the progression of renal fibrosis, both in vitro and in vivo.
- The findings suggest a significant role for endostatin in the pathogenesis of tubulointerstitial injury in progressive kidney diseases.
