Inhibition of MiR-30d-5p Promotes Mitochondrial Autophagy and Alleviates Podocyte Injury under High Glucose

Ying Cai; Sheng Chen; Xiaoli Jiang; Qiyuan Wu; Bei Guo; Fang Wang

Authors

Ying Cai Department of Nephrology, Lihuili Hospital, Ningbo Medical Center, 315000 Ningbo, Zhejiang, China Author
Sheng Chen Department of Nephrology, Lihuili Hospital, Ningbo Medical Center, 315000 Ningbo, Zhejiang, China Author
Xiaoli Jiang Department of Nephrology, Lihuili Hospital, Ningbo Medical Center, 315000 Ningbo, Zhejiang, China Author
Qiyuan Wu Department of Nephrology, Lihuili Hospital, Ningbo Medical Center, 315000 Ningbo, Zhejiang, China Author
Bei Guo Department of Nephrology, Lihuili Hospital, Ningbo Medical Center, 315000 Ningbo, Zhejiang, China Author
Fang Wang Department of Nephrology, Lihuili Hospital, Ningbo Medical Center, 315000 Ningbo, Zhejiang, China Author

Keywords:

MiR-30d-5p, mitochondrial function, autophagy, high glucose, podocyte

Abstract

Objective: To study the role of miR-30d-5p in high glucose-induced podocyte injury. Methods: Podocytes were hyperglycated with 30 mM glucose, transfected with miR-30d-5p inhibitor and mimic, and then treated with 1 mg/mL 3-MA. The transfection efficiency of miR-30d-5p and the expressions of mitochondrial autophagy related genes (ATG5, PINK1 and PARK2) were detected by qRT-PCR. Apoptosis was detected by flow cytometry. The expressions of LC3I, LC3II, P62, ATG5, PINK1 and PARK2 were measured by Western blotting. JC-1 was used as a fluorescent probe to detect the mitochondrial membrane potential, and Adenosine Triphosphate (ATP) content in cells was quantified by relevant kits. Results: In the high glucose-induced podocytes, miR-30d-5p and P62 expressions were upregulated, while ATG5, PINK1, PARK2 and LC3II/LC3I expression levels were downregulated. MiR-30d-5p inhibitor reversed the effect of high glucose on the expressions of ATG5, PINK1, PARK2, LC3I, LC3II and P62. High glucose induced loss of mitochondrial membrane potential and ATP content in podocytes, and increased membrane potential and ATP content were observed after inhibition of miR-30d-5p. Conclusion: Inhibition of miR-30d-5p may enhance mitochondrial autophagy by promoting the expressions of ATG5 and PINK1/2, and subsequently alleviate high glucose-induced podocyte damage.

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