miRNA-483-5p Targets HDCA4 to Regulate Renal Tubular Damage in Diabetic Nephropathy
Lu Liu1, Huanzhen Chen2, Jie Yun3
1Department of Endocrinology, Seventh People's Hospital of Shanghai University of TCM, 200137, Shanghai, China.
Abstract:
This study was designed to evaluate the diagnostic value of miR-483-5p in diabetic nephropathy (DN), and its effect and mechanism on apoptosis and inflammation of human proximal renal tubular cells (HK2) induced by high glucose (HG). Thirty healthy controls, 30 types 2 diabetes mellitus (T2DM) patients, and 28 DN patients were enrolled. miR-483-5p mRNA levels in serum were analyzed by RT-qPCR assays. The receiver operating characteristic curve (ROC) was used to analyze the diagnostic value of miR-483-5p in DN. HK2 cells were induced by HG to establish an in vitro study model. CCK-8 and flow cytometry was used to detect cell viability, apoptosis, and reactive oxygen species (ROS) generation. Inflammation levels were measured by ELISA. Luciferase reporter assay was used to detect target genes of miR-483-5p. miR-483-5p was decreased in DN patients. The decreased level of miR-483-5p was positively correlated with estimated glomerular filtration rate (eGFR) and negatively correlated with proteinuria. miR-483-5p can significantly distinguish DN patients from healthy controls and T2DM and has a high diagnostic value. miR-483-5p decreased in HK2 cells induced by HG, and overexpression of miR-483-5p reversed HG-induced decreased cell activity, increased apoptosis, ROS production, and inflammation. Histone deacetylase 4 (HDCA4) was markedly increased in DN patients and HG-induced HK2 cells. miR-483-5p directly targeted HDCA4, and increasing miR-483-5p inhibited HDCA4 increased in HG-induced HK2. In conclusion, the results indicate that reduction of miR-483-5p has a high diagnostic value in DN, and overexpression of miR-483-5p has a certain protective effect on HK2 cells induced by HG by targeting HDCA4.
Insights
Reduced miR-483-5p levels are a valuable diagnostic marker for diabetic nephropathy (DN). Restoring miR-483-5p protects kidney cells from high glucose damage by targeting HDCA4.
Area of Science:
- Molecular Biology
- Biochemistry
- Nephrology
Background:
- Diabetic nephropathy (DN) is a major complication of diabetes, characterized by kidney damage.
- Biomarkers are needed for early diagnosis and understanding the molecular mechanisms of DN.
- MicroRNAs (miRNAs) play crucial roles in cellular processes and disease development.
Purpose of the Study:
- To evaluate the diagnostic potential of miR-483-5p in diabetic nephropathy (DN).
- To investigate the role of miR-483-5p in high glucose-induced apoptosis and inflammation in human proximal renal tubular cells (HK2).
- To elucidate the underlying molecular mechanism involving miR-483-5p and its target gene.
Main Methods:
- Serum miR-483-5p levels were quantified using RT-qPCR in healthy controls, type 2 diabetes mellitus (T2DM) patients, and DN patients.
- Diagnostic value was assessed using receiver operating characteristic (ROC) curve analysis.
- In vitro studies utilized high glucose (HG)-induced HK2 cells to examine cell viability, apoptosis, reactive oxygen species (ROS), inflammation, and target gene interactions via luciferase reporter assays.
Main Results:
- Serum miR-483-5p levels were significantly decreased in DN patients compared to controls and T2DM patients.
- Decreased miR-483-5p levels correlated positively with estimated glomerular filtration rate (eGFR) and negatively with proteinuria, indicating diagnostic value for DN.
- Overexpression of miR-483-5p in HG-induced HK2 cells attenuated cell damage, apoptosis, ROS production, and inflammation, identifying Histone deacetylase 4 (HDCA4) as a direct target.
Conclusions:
- Reduced miR-483-5p in serum demonstrates high diagnostic value for identifying diabetic nephropathy.
- miR-483-5p exerts protective effects against high glucose-induced kidney tubular cell injury.
- The protective mechanism involves the direct targeting of HDCA4 by miR-483-5p, suggesting a novel therapeutic pathway for DN.
