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ClC-5 alleviates renal fibrosis in unilateral ureteral obstruction mice

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Abstract

Renal fibrosis is the major feature of end-stage renal disease with high mortality. Chloride (Cl−) moving along Cl− channels has been suggested to play to an important role in renal function. This study aims to investigate the role of ClC-5 in renal fibrosis in unilateral ureteral occlusion (UUO) mice. C57BL/6 mice received UUO surgery followed by delivery of adeno-associated virus encoding ClC-5 cDNA (AAVClC-5). Western blotting, real-time PCR and histological analysis were used to investigate the effects of ClC-5 on renal fibrosis and underlying mechanisms. The expression of ClC-5 was significantly decreased in renal cortex of UUO mice and transforming growth factor-β1 (TGF-β1)-stimulated HK2 cells. Overexpression of ClC-5 in vivo markedly ameliorated UUO-induced renal injury and fibrosis. The increased expressions of plasminogen activator inhibitor type 1, connective tissue growth factor, collagen III and collagen IV were also inhibited by ClC-5 upregulation. Moreover, UUO-induced immune cell infiltration and inflammatory cytokines release were attenuated in mice infected with AAVClC-5. In addition, the in vivo and in vitro results showed that ClC-5 overexpression prevented epithelial-to-mesenchymal transition (EMT), concomitantly with a restoration of E-cadherin expression and a decrease of vimentin, α-SMA and S100A4 expressions. Furthermore, ClC-5 overexpression inhibited UUO- or TGF-β1-induced increase in nuclear factor kappa B (NF-κB) acetylation and matrix metalloproteinases-9 (MMP-9) expression. However, downregulation of ClC-5 in HK2 cells further potentiated TGF-β1-induced EMT and increase in NF-κB acetylation and MMP-9 expression. ClC-5 upregulation ameliorates renal fibrosis via inhibiting NF-κB/MMP-9 pathway signaling activation, suggesting that ClC-5 may be a novel therapeutic target for treating renal fibrosis and chronic kidney disease.

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Abbreviations

UUO:

Unilateral ureteral occlusion

AAV:

Adeno-associated virus

TGF-β1:

Transforming growth factor-β1

NF-κB:

Nuclear factor kappa B

MMP:

Matrix metalloproteinases-9

ECM:

Extracellular matrix

EMT:

Epithelial-to-mesenchymal transition

PAI-1:

Plasminogen activator inhibitor type 1

CTGF:

Connective tissue growth factor

IL-1β:

Interleukin-1β

IL-6:

Interleukin-6

MCP-1:

Monocyte chemoattractant protein-1

ICAM-1:

Intercellular adhesion molecule-1

TNF-α:

Tumor necrosis factor-α

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Authors and Affiliations

  1. Department of Nephrology, Gansu Provincial Hospital of Traditional Chinese Medicine, No. 418 Guazhou Road, Lanzhou, 730050, Gansu, China

    Shi-Xia Yang & Hui-Ling Bai

  2. Department of Neurology, Lanzhou University Second Hospital, Lanzhou, 730030, Gansu, China

    Zheng-Chang Zhang

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Correspondence to Shi-Xia Yang.

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All animal experiments were carried out according to the institutional guidelines from the Principles of Laboratory Animal Care of Gansu Provincial Hospital of Traditional Chinese Medicine.

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Yang, SX., Zhang, ZC. & Bai, HL. ClC-5 alleviates renal fibrosis in unilateral ureteral obstruction mice. Human Cell 32, 297–305 (2019). https://doi.org/10.1007/s13577-019-00253-5

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