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MiR-449a downregulation alleviates the progression of renal interstitial fibrosis by mediating the KLF4/MFN2 axis

  • Nephrology - Original Paper
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Abstract

Background

Renal interstitial fibrosis (RIF) seriously threatens the health of individuals. MiRNAs regulate the progression of fibrosis. Nevertheless, the detailed function of miR-449a in RIF is largely unknown.

Methods

In vitro and in vivo models of RIF were developed to evaluate the function of miR-449a. The relationship among miR-449a, KLF4, and MFN2 was explored using a dual-luciferase reporter assay and chromatin immunoprecipitation. Additionally, the pathological changes in the mice were detected using Masson staining. The mRNA and protein expressions were assessed using quantitative reverse transcription polymerase chain reaction and western blot, respectively.

Results

TGF-β1 downregulated the expressions of KLF4 and MFN2 in TCMK-1 cells, but upregulated the level of miR-449a. The downregulation of miR-449a significantly inhibited TGF-β1-induced upregulation of fibrotic proteins in TCMK-1 cells. Meanwhile, miR-449a directly targeted KLF4. Moreover, KLF4 overexpression activated MFN2 transcription and reversed TGF-β1-induced fibrosis by positively regulating MFN2. Furthermore, the downregulation of miR-449a could obviously alleviate the symptoms of RIF in mice with unilateral ureteral obstruction.

Conclusion

MiR-449a downregulation attenuated the development of RIF by mediating the KLF4/MFN2 axis. Therefore, miR-449a might act as a target in treating RIF.

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Data availability

All data generated or analyzed during this study are included in this article. The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

MFN2:

Mitofusin 2

TGF-β:

Transforming growth factor-β

RIF:

Renal interstitial fibrosis

CKD:

Chronic kidney disease

Smad:

Drosophila mothers against decapentaplegic protein

KLF4:

Krüppel-like factor 4

rAAV:

Recombinant adeno-associated virus

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Funding

This work was supported by National Natural Science Foundation of China (81760130), National Natural Science Foundation of China (82160143) and The Engineering Research Center of Kidney Disease in Jiangxi Province (20164BCD40095).

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Authors and Affiliations

  1. Department of Nephrology, The Second Affiliated Hospital of Nanchang University, No. 1, Minde Road, Nanchang, 330006, Jiangxi Province, China

    Xiang-Dong Fang, Yan-Xia Chen, Ben Ke, Shu-Ying Zhu, Chu-Qiao Fan & Wei-Ping Tu

  2. Department of Clinical Pharmacology, The Second Affiliated Hospital of Nanchang University, Nanchang, 330006, Jiangxi Province, China

    Jia-Ke He

  3. Department of Cardiovascular Medicine, The Second Affiliated Hospital of Nanchang University, No. 1, Minde Road, Nanchang, 330006, Jiangxi Province, China

    Ping Li

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Contributions

W-PT, X-DF: conception and design of the work. J-KH, Y-XC: data acquisition, drafting the work. BK, S-YZ: data analysis and data interpretation. C-QF, PL: revising it critically for important intellectual content. All author approved the final version of the manuscript to be published.

Corresponding authors

Correspondence to Wei-Ping Tu or Ping Li.

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Not applicable.

Ethical approval

The experimental protocol was approved by the Ethics Committee for Animal Experimentation of The Second Affiliated Hospital of Nanchang University, and it was in line with the Guidelines of Animal Experimentation of The Second Affiliated Hospital of Nanchang University.

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11255_2023_3503_MOESM1_ESM.tif

Supplementary file1 Supplementary Figure 1. Tenascin-C significantly upregulated the expression of miR-449a in TCMK-1 cells. TCMK-1 cells were added with Tenascin-C (50 ng/mL). (A) The expression of miR-449a in TCMK-1 cells was detected using RT-qPCR. (B) α-SMA, fibronectin, and collagen IV levels in TCMK-1 cells were assessed using western blot. β-actin was applied for normalization. *p < 0.05, **p < 0.01, ***p < 0.001 (TIF 486 KB)

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Fang, XD., He, JK., Chen, YX. et al. MiR-449a downregulation alleviates the progression of renal interstitial fibrosis by mediating the KLF4/MFN2 axis. Int Urol Nephrol 55, 1837–1846 (2023). https://doi.org/10.1007/s11255-023-03503-6

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