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Apamin inhibits renal fibrosis via suppressing TGF-β1 and STAT3 signaling in vivo and in vitro

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Abstract

Renal fibrosis is a progressive and chronic process that influences kidneys with chronic kidney disease (CKD), irrespective of cause, leading to irreversible failure of renal function and end-stage kidney disease. Among the signaling related to renal fibrosis, transforming growth factor-β1 (TGF-β1) signaling is a major pathway that induces the activation of myofibroblasts and the production of extracellular matrix (ECM) molecules. Apamin, a component of bee venom (BV), has been studied in relation to various diseases. However, the effect of apamin on renal interstitial fibrosis has not been investigated. The aim of this study was to estimate the beneficial effect of apamin in unilateral ureteral obstruction (UUO)-induced renal fibrosis and TGF-β1-induced renal fibroblast activation. This study revealed that obstructive kidney injury induced an inflammatory response, tubular atrophy, and ECM accumulation. However, apamin treatment suppressed the increased expression of fibrotic-related genes, including α-SMA, vimentin, and fibronectin. Administration of apamin also attenuated the renal tubular cells injury and tubular atrophy. In addition, apamin attenuated fibroblast activation, ECM synthesis, and inflammatory cytokines such as TNF-α, IL-1β, and IL-6 by suppressing the TGF-β1-canonical and non-canonical signaling pathways. This study showed that apamin inhibits UUO-induced renal fibrosis in vivo and TGF-β1-induced renal fibroblasts activation in vitro. Apamin inhibited the inflammatory response, tubular atrophy, ECM accumulation, fibroblast activation, and renal interstitial fibrosis through suppression of TGF-β1/Smad2/3 and STAT3 signaling pathways. These results suggest that apamin might be a potential therapeutic agent for renal fibrosis.

Key messages

  • UUO injury can induce renal dysfunction; however, apamin administration prevents renal failure in UUO mice.

  • Apamin inhibited renal inflammatory response and ECM deposition in UUO-injured mice.

  • Apamin suppressed the activation of myofibroblasts in vivo and in vitro.

  • Apamin has the anti-fibrotic effect on renal fibrosis via regulation of TGF-β1 canonical and non-canonical signaling.

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Availability of data and materials

The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

α-SMA:

α-smooth muscle actin

BUN:

Blood urea nitrogen

BV:

Bee venom

CKD:

Chronic kidney disease

ECM:

Extracellular matrix

FSP-1:

Fibroblast-specific marker-1

IL-1β:

Interleukin-1β

MCP-1:

Monocyte chemoattractant protein-1

NGAL:

Neutrophil gelatinase-associated lipocalin

SK channel:

Small conductance Ca2+-activated K+ channel

STAT3:

Signal transducer and activator of transcription 3

TβRI:

TGF-β type I receptor

TβRII:

TGF-β type II receptor

TGF-β1:

Transforming growth factor-β1

UUO:

Unilateral ureteral obstruction

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Funding

This work was supported by the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture, Forestry (IPET) through the Useful Agricultural Life Resources Industry Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (grant number 120040-02-1).

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, Catholic University of Daegu, 33, Duryugongwon-ro 17-gil, Nam-gu, 42472, Daegu, Republic of Korea

    Mi-Gyeong Gwon, Hyun-** An, Hyemin Gu, Young-Ah Kim & Kwan-Kyu Park

  2. National Academy of Agricultural Science, Jeonjusi, Jeonbuk, 54875, Republic of Korea

    Sang Mi Han

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  1. Mi-Gyeong Gwon
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  2. Hyun-** An
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  3. Hyemin Gu
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Contributions

M.-G.G. and K.-K.P. participated in the design of the study. M.-G.G., H.-J.A., and H.G. performed the experiments. Y.-A.K. and S.M.H. performed the data analysis. M.-G.G. and K.-K.P. drafted the manuscripts. All authors discussed, revised, and approved the final manuscript.

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Correspondence to Kwan-Kyu Park.

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All protocols for animal study were approved by the Institutional Animal Care and Use Committee of the Catholic University of Daegu (EXP-IRB number: DCIAFCR-160705-6-Y). All animal experiments have been performed in accordance with the ethical standards outlined in the Best Practice Guidelines on Publishing Ethics.

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Supplementary Information

Fig. S1

Establishment of the renal fibrosis models and schematic diagram of apamin treatment. a Establishment of the renal fibrosis model and schematic diagram of apamin administration. Apamin treatment at a concentration of 0.5 mg/kg was given via intraperitoneal injection twice a week (PNG 269 kb)

High resolution image (TIF 8857 kb)

Fig. S2

Apamin attenuated the renal tubular injury in UUO mice. a The protein expressions of E-cadherin and NGAL, the markers of tubular injury, were analyzed with western blotting analysis. The quantitative graphs show protein expression of b E-cadherin and c NGAL normalized with GAPDH. *p < 0.05 compared to the normal control group; †p < 0.05 compared to the UUO group (PNG 617 kb)

High resolution image (TIF 13806 kb)

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Gwon, MG., An, HJ., Gu, H. et al. Apamin inhibits renal fibrosis via suppressing TGF-β1 and STAT3 signaling in vivo and in vitro. J Mol Med 99, 1265–1277 (2021). https://doi.org/10.1007/s00109-021-02087-x

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