Inhibition of miR-30d-5p promotes mitochondrial autophagy and alleviates high glucose-induced injury in podocytes.
10.3724/zdxbyxb-2024-0504
- Author:
Ying CAI
1
,
2
;
Sheng CHEN
3
;
Xiaoli JIANG
3
;
Qiyuan WU
3
;
Bei GUO
3
;
Fang WANG
2
,
4
Author Information
1. Department of Nephrology, Ningbo Medical Center Lihuili Hospital, Ningbo 315000, Zhejiang Province, China. caiying_cy1@
2. com.
3. Department of Nephrology, Ningbo Medical Center Lihuili Hospital, Ningbo 315000, Zhejiang Province, China.
4. Department of Nephrology, Ningbo Medical Center Lihuili Hospital, Ningbo 315000, Zhejiang Province, China. wangffff222@
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Autophagy;
Diabetic nephropathy;
High glucose;
MicroRNA-30d-5p;
Mitochondrial function;
Podocytes
- MeSH:
Podocytes/drug effects*;
MicroRNAs/metabolism*;
Glucose/adverse effects*;
Autophagy/drug effects*;
Animals;
Mice;
Autophagy-Related Protein 5/genetics*;
Apoptosis/drug effects*;
Mitochondria/metabolism*;
Ubiquitin-Protein Ligases/genetics*;
Membrane Proteins/genetics*;
Membrane Potential, Mitochondrial/drug effects*;
Microtubule-Associated Proteins/genetics*;
Protein Kinases
- From:
Journal of Zhejiang University. Medical sciences
2024;53(6):756-764
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES:To study the role of microRNA (miR)-30d-5p in high glucose-induced podocyte injury.
METHODS:Podocytes were hyperglycated with 30 mmol/L glucose, transfected with miR-30d-5p inhibitor and mimic, and then treated with 1 mg/mL 3-methyladenine (3-MA). The transfection efficiency of miR-30d-5p was quantified by reverse transcription PCR. Apoptosis was detected by flow cytometry. The expressions of nephrin, microtubule-associated protein light chain (LC) 3Ⅱ/LC3Ⅰ, P62, autophagy-related gene (ATG) 5, PTEN induced putative kinase (PINK) 1 and Parkin gene (PARK2) were detected by Western blotting. The mito-chondrial membrane potential was detected by JC-1 fluorescent probe, and adenosine triphosphate (ATP) content in cells was detected by relevant kits.
RESULTS:Under high glucose induction, podocyte apoptosis increased, miR-30d-5p and P62 expressions were upregulated, while nephrin, ATG5, PINK1, PARK2 and LC3Ⅱ/LC3Ⅰ expressions decreased (all P<0.01). MiR-30d-5p inhibitor reversed the effect of high glucose on apoptosis, and the expression of ATG5, PINK1, PARK2, nephrin, LC3Ⅱ/LC3Ⅰ and P62 (all P<0.01). High glucose induced loss of mitochondrial membrane potential and ATP content in podocytes, while inhibition of miR-30d-5p increased them. Autophagy inhibitors 3-MA and miR-30d-5p mimics reversed the effects of miR-30d-5p inhibition on apoptosis, autophagy and mitochondrial function of podocytes induced by high glucose (all P<0.05).
CONCLUSIONS:Inhibition of miR-30d-5p may promote mitochondrial autophagy (mitophagy) by promoting the expression of ATG5, PINK1, PARK2 and alleviating high glucose-induced podocyte damage.
