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Updated: Dec 16, 2025
A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model
Published on: September 11, 2020
miR-10a overexpression aggravates renal ischemia-reperfusion injury associated with decreased PIK3CA expression
Dongsheng Xu1, Wenjun Li2, Tao Zhang3
1Department of Kidney Transplantation, The Second Hospital of Shandong University, Jinan, 250033, China.
Background:
To investigate the effect of miR-10a on renal tissues with ischemia reperfusion (I/R) injury in rats and to explore the underlying mechanisms of the effect of miR-10a on hypoxia-reoxygenation in HK-2 cells.
Methods:
MiR-10a level was measured in the renal tissues of rats with I/R rats using RT-PCR. In order to research the role of miR-10a in renal tissues, an miR-10 agonist and an miR-10a antagonist were used to treat I/R-injured rats. Levels of serum creatinine and blood urea nitrogen, renal histopathology, and levels of cell apoptosis were analyzed separately in renal tissues from the rats. Phosphatidylinositol 3-kinase (PI3K)/Akt pathway related proteins were measured by Western blotting. In addition, HK-2 cells were cultured in order to study the mechanism of action of miR-10a in the hypoxia-reoxygenation model being studied. Finally, the dual luciferase reporter gene assay was used to confirm that the PI3K p100 catalytic subunit α (PIK3CA) gene was targeted by miR-10a.
Results:
After renal I/R injury in rats, miR-10a expression increased significantly (p < 0.05). Injection of miR-10a agonist significantly aggravated the renal injury and raised the level of cell apoptosis in the renal tissues of I/R-injured rats (p < 0.05). However, administration of miR-10a antagonist led to obvious improvement of the renal injury, decreased renal cell apoptosis, and inhibited PI3K/Akt pathway activity (p < 0.05). In in vitro experiments, the negative relationship between PIK3CA and miR-10a levels was confirmed. Furthermore, overexpression of miR-10a significantly decreased the proliferation of HK-2 cells, and increased cell apoptosis via up-regulation of the PI3K/Akt pathway (p < 0.05).
Conclusion:
The aggravation of renal I/R injury by miR-10a was associated with a decrease in the activity of PIK3CA/PI3K/Akt pathway.
Insights
MicroRNA-10a (miR-10a) exacerbates renal ischemia-reperfusion (I/R) injury by inhibiting the PI3K/Akt pathway. Antagonizing miR-10a protects renal tissues and reduces apoptosis, suggesting therapeutic potential for I/R injury.
Area of Science:
- Nephrology
- Molecular Biology
- Cell Biology
Background:
- Renal ischemia-reperfusion (I/R) injury is a significant clinical challenge.
- The role of microRNAs in regulating renal injury remains incompletely understood.
- MicroRNA-10a (miR-10a) has emerged as a potential regulator in cellular stress responses.
Purpose of the Study:
- To investigate the impact of miR-10a on renal I/R injury in a rat model.
- To elucidate the underlying molecular mechanisms of miR-10a in hypoxia-reoxygenation injury in HK-2 cells.
- To explore the potential of targeting miR-10a for therapeutic intervention in renal injury.
Main Methods:
- Real-time PCR (RT-PCR) to quantify miR-10a levels in renal tissues.
- Administration of miR-10a agonist and antagonist in I/R-injured rats.
- Assessment of serum creatinine, blood urea nitrogen, renal histopathology, and apoptosis.
- Western blotting to analyze Phosphatidylinositol 3-kinase (PI3K)/Akt pathway proteins.
- Dual luciferase reporter gene assay to identify PIK3CA as a miR-10a target.
- In vitro experiments using HK-2 cells under hypoxia-reoxygenation conditions.
Main Results:
- miR-10a expression significantly increased following renal I/R injury.
- miR-10a agonist aggravated renal injury and apoptosis, while antagonist improved injury and reduced apoptosis.
- miR-10a antagonist inhibited PI3K/Akt pathway activity.
- A negative correlation between PIK3CA and miR-10a levels was confirmed.
- Overexpression of miR-10a reduced HK-2 cell proliferation and increased apoptosis via the PI3K/Akt pathway.
Conclusions:
- miR-10a plays a detrimental role in renal I/R injury.
- The pro-injury effect of miR-10a is mediated through the suppression of the PIK3CA/PI3K/Akt pathway.
- Targeting miR-10a offers a potential therapeutic strategy for mitigating renal I/R injury.
