ASN's Mission

To create a world without kidney diseases, the ASN Alliance for Kidney Health elevates care by educating and informing, driving breakthroughs and innovation, and advocating for policies that create transformative changes in kidney medicine throughout the world.

learn more

Contact ASN

1401 H St, NW, Ste 900, Washington, DC 20005

email@asn-online.org

202-640-4660

The Latest on Twitter

Kidney Week

Abstract: FR-PO401

Disruption of Renal Tubular Mitochondrial Quality Control by KCa3.1 in Diabetic Nephropathy

Session Information

Category: Diabetic Kidney Disease

  • 601 Diabetic Kidney Disease: Basic

Authors

  • Huang, Chunling, kolling institute, the University of Sydney, St. Leonards, New South Wales, Australia
  • Yi, Hao, kolling institute, the University of Sydney, St. Leonards, New South Wales, Australia
  • Shi, Ying, kolling institute, the University of Sydney, St. Leonards, New South Wales, Australia
  • Cao, Qinghua, kolling institute, the University of Sydney, St. Leonards, New South Wales, Australia
  • Chen, Xinming, kolling institute, the University of Sydney, St. Leonards, New South Wales, Australia
  • Pollock, Carol A., kolling institute, the University of Sydney, St. Leonards, New South Wales, Australia
Background

Mitochondrial dysfunction is involved in the pathogenesis of diabetic nephropathy. Mitochondrial quality control is characterized by repairing of mitochondrial damage through mitophagy and fission/fusion. It has been shown that blockade of KCa3.1, a potassium channel, ameliorates diabetic renal fibrosis and KCa3.1 activation contributes to dysfunctional tubular autophagy in diabetic nephropathy through PI3K/Akt/mTOR signaling pathways. However, the role of KCa3.1 in mitochondrial quality control is not yet known.Therefore, the aim of the study is to identify the role of KCa3.1 in mitochondrial quality control in diabetic nephropathy.

Methods

In vitro human proximal tubular cells (HK2 cells) transfected with scrambled siRNA or KCa3.1 siRNA were exposed to TGF-β1 for 48h. Mitochondrial function and mitochondrial ROS (mtROS) production were assessed. In vivo, diabetes was induced in KCa3.1+/+ and KCa3.1-/- mice by streptozotocin injection. The pro-fission protein dynamin-related protein 1 (Drp1) and pro-fussion protein mitofusin 2 (Mfn2) as well as BCL2 interacting protein 3 (BNIP3) (a mitophagy regulator) were examined by western blotting in HK2 cells and mice kidneys.

Results

The in vitro results showed that TGF-β1 significantly inhibited mitochondrial ATP production rate, compared to the controls, which were significantly reversed by KCa3.1 siRNA in HK2 cells. KCa3.1 gene silencing inhibited TGF-β1-induced significant increase in MitoSOX Red fluorescence in HK2 cells. TGF-β1 significantly increased the expression of Drp1 and BNIP3 in HK2 cells, which were attenuated by KCa3.1 gene silencing. The expression of Mfn2 was not overtly apparent on TGF-β1 stimulation. Consistently, the in vivo results showed significantly increased Drp1 and BNIP3 expression in diabetic KCa3.1 +/+ mice, which were significantly reduced in diabetic KCa3.1-/- mice.

Conclusion

KCa3.1 mediates dysregulation of mitochondrial quality control in diabetic nephropathy.