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Circ_0060077 Knockdown Alleviates High-Glucose-Induced Cell Apoptosis, Oxidative Stress, Inflammation and Fibrosis in HK-2 Cells via miR-145-5p/VASN Pathway


The involvement of circular RNAs (circRNAs) in the progression of diabetic nephropathy (DN) has been reported. However, the functions of circ_0060077 in DN remain unclear. HK-2 cells were treated with high glucose (HG) to establish DN cell model. Quantitative real-time polymerase chain reaction (qRT-PCR) was proceeded to determine the levels of circ_0060077, microRNA-145-5p (miR-145-5p) and vasorin (VASN). Cell counting kit-8 (CCK-8) assay, 5-ethynyl-2’-deoxyuridine (EdU) assay and colony formation assay were conducted to assess cell proliferation ability. Flow cytometry analysis was employed for cell apoptosis. The oxidative stress level was evaluated by commercial kits. Enzyme-linked immunosorbent assay (ELISA) was adopted to examine the concentrations of inflammatory factors. Western blot assay was utilized for protein levels. Dual-luciferase reporter assay and RNA pull-down assay were manipulated to analyze the relationships among circ_0060077, miR-145-5p and VASN. Circ_0060077 level was increased in DN patients and HG-stimulated HK-2 cells. Circ_0060077 knockdown ameliorated the inhibitory effect of HG on HK-2 cell proliferation and the promotional effects on cell apoptosis, oxidative stress, inflammation and fibrosis. MiR-145-5p was identified as the target for circ_0060077 and miR-145-5p inhibition ameliorated the effect of circ_0060077 silencing on HG-induced HK-2 cell injury. Moreover, miR-145-5p directly bound to VASN. Overexpression of miR-145-5p facilitated cell proliferation and repressed apoptosis, oxidative injury, inflammation and fibrosis in HG-induced HK-2 cells by targeting VASN. Circ_0060077 silencing protected HK-2 cells from HG-induced damage by regulating miR-145-5p/VASN axis.

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  1. Magee, C., D.J. Grieve, C.J. Watson, and D.P. Brazil. 2017. Diabetic Nephropathy: A Tangled Web to Unweave. Cardiovascular Drugs and Therapy 31 (5–6): 579–592.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Rossing P. 2006. Prediction, progression and prevention of diabetic nephropathy. The Minkowski Lecture 2005. Diabetologia 49 (1): 11–19.

  3. Loeffler, I., and G. Wolf. 2015. Epithelial-to-Mesenchymal Transition in Diabetic Nephropathy: Fact or Fiction? Cells 4 (4): 631–652.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Ni, W.J., L.Q. Tang, and W. Wei. 2015. Research progress in signalling pathway in diabetic nephropathy. Diabetes/Metabolism Research and Reviews 31 (3): 221–233.

    Article  PubMed  Google Scholar 

  5. Alicic, R.Z., M.T. Rooney, and K.R. Tuttle. 2017. Diabetic Kidney Disease: Challenges, Progress, and Possibilities. Clinical Journal of the American Society of Nephrology 12 (12): 2032–2045.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Ebbesen, K.K., T.B. Hansen, and J. Kjems. 2017. Insights into circular RNA biology. RNA Biology 14 (8): 1035–1045.

    Article  PubMed  Google Scholar 

  7. Hansen, T.B., T.I. Jensen, B.H. Clausen, J.B. Bramsen, B. Finsen, C.K. Damgaard, and J. Kjems. 2013. Natural RNA circles function as efficient microRNA sponges. Nature 495 (7441): 384–388.

    CAS  Article  PubMed  Google Scholar 

  8. Han, B., J. Chao, and H. Yao. 2018. Circular RNA and its mechanisms in disease: From the bench to the clinic. Pharmacology & Therapeutics 187: 31–44.

    CAS  Article  Google Scholar 

  9. Zhao, L., H. Chen, Y. Zeng, K. Yang, R. Zhang, Z. Li, T. Yang, and H. Ruan. 2021. Circular RNA circ_0000712 regulates high glucose-induced apoptosis, inflammation, oxidative stress, and fibrosis in (DN) by targeting the miR-879-5p/SOX6 axis. Endocrine Journal.

    Article  PubMed  Google Scholar 

  10. An, L., D. Ji, W. Hu, J. Wang, X. Jin, Y. Qu, and N. Zhang. 2020. Interference of Hsa_circ_0003928 Alleviates High Glucose-Induced Cell Apoptosis and Inflammation in HK-2 Cells via miR-151-3p/Anxa2. Diabetes, Metabolic Syndrome and Obesity 13: 3157–3168.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  11. Yang, L., X. Han, C. Zhang, C. Sun, S. Huang, W. Xiao, Y. Gao, Q. Liang, F. Luo, W. Lu, J. Fu, and Y. Zhou. 2020. Hsa_circ_0060450 Negatively Regulates Type I Interferon-Induced Inflammation by Serving as miR-199a-5p Sponge in Type 1 Diabetes Mellitus. Frontiers in Immunology 11: 576903.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. Tang, J., D. Yao, H. Yan, X. Chen, L. Wang, and H. Zhan. 2019. The Role of MicroRNAs in the Pathogenesis of Diabetic Nephropathy. International Journal of Endocrinology 2019: 8719060.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. Liu, L., H. Chen, J. Yun, L. Song, X. Ma, S. Luo, and Y. Song. 2021. miRNA-483-5p Targets HDCA4 to Regulate Renal Tubular Damage in Diabetic Nephropathy. Hormone and Metabolic Research.

    Article  PubMed  Google Scholar 

  14. Chen, X., L. Gu, X. Cheng, J. Xing, and M. Zhang. 2021. MiR-17-5p downregulation alleviates apoptosis and fibrosis in high glucose-induced human mesangial cells through inactivation of Wnt/beta-catenin signaling by targeting KIF23. Environmental Toxicology.

    Article  PubMed  Google Scholar 

  15. Wei, B., Y.S. Liu, and H.X. Guan. 2020. MicroRNA-145-5p attenuates high glucose-induced apoptosis by targeting the Notch signaling pathway in podocytes. Experimental and Therapeutic Medicine 19 (3): 1915–1924.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  16. Liang, W., B. Guo, J. Ye, H. Liu, W. Deng, C. Lin, X. Zhong, and L. Wang. 2019. Vasorin stimulates malignant progression and angiogenesis in glioma. Cancer Science 110 (8): 2558–2572.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  17. Bhandari, A., Y. Guan, E. Xia, Q. Huang, and Y. Chen. 2019. VASN promotes YAP/TAZ and EMT pathway in thyroid carcinogenesis in vitro. American Journal of Translational Research 11 (6): 3589–3599.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Cui F.L., A.N., Mahmud, Z.P. Xu, Z.Y. Wang, and J.P. Hu. 2020. VASN promotes proliferation of prostate cancer through the YAP/TAZ axis. European Review for Medical and Pharmacological Sciences 24 (12): 6589–6596.

  19. Ahn, J.M., B.G. Kim, M.H. Yu, I.K. Lee, and J.Y. Cho. 2010. Identification of diabetic nephropathy-selective proteins in human plasma by multi-lectin affinity chromatography and LC-MS/MS. Proteomics. Clinical Applications 4 (6–7): 644–653.

    CAS  Article  PubMed  Google Scholar 

  20. Jin, J., H. Sun, C. Shi, H. Yang, Y. Wu, W. Li, Y.H. Dong, L. Cai, and X.M. Meng. 2020. Circular RNA in renal diseases. Journal of Cellular and Molecular Medicine 24 (12): 6523–6533.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Zhang, J.R., and H.J. Sun. 2020. Roles of circular RNAs in diabetic complications: From molecular mechanisms to therapeutic potential. Gene 763: 145066.

    CAS  Article  PubMed  Google Scholar 

  22. Meng, Q., X. Zhai, Y. Yuan, Q. Ji, and P. Zhang. 2020. lncRNA ZEB1-AS1 inhibits high glucose-induced EMT and fibrogenesis by regulating the miR-216a-5p/BMP7 axis in diabetic nephropathy. Brazilian Journal of Medical and Biological Research 53 (4): e9288.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  23. Wang, J., and S.M. Zhao. 2021. LncRNA-antisense non-coding RNA in the INK4 locus promotes pyroptosis via miR-497/thioredoxin-interacting protein axis in diabetic nephropathy. Life Sciences 264: 118728.

    CAS  Article  PubMed  Google Scholar 

  24. Liu, H., X. Wang, Z.Y. Wang, and L. Li. 2020. Circ_0080425 inhibits cell proliferation and fibrosis in diabetic nephropathy via sponging miR-24-3p and targeting fibroblast growth factor 11. Journal of Cellular Physiology 235 (5): 4520–4529.

    CAS  Article  PubMed  Google Scholar 

  25. Tang, B., W. Li, T.T. Ji, X.Y. Li, X. Qu, L. Feng, and S. Bai. 2020. Circ-AKT3 inhibits the accumulation of extracellular matrix of mesangial cells in diabetic nephropathy via modulating miR-296-3p/E-cadherin signals. Journal of Cellular and Molecular Medicine 24 (15): 8779–8788.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  26. Li, J., X. Jiang, L. Duan, and W. Wang. 2019. Long non-coding RNA MEG3 impacts diabetic nephropathy progression through sponging miR-145. American Journal of Translational Research 11 (10): 6691–6698.

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Liu B., L. Qiang, G.D. Wang, Q. Duan, and J. Liu. 2019. LncRNA MALAT1 facilities high glucose induced endothelial to mesenchymal transition and fibrosis via targeting miR-145/ZEB2 axis. European Review for Medical and Pharmacological Sciences 23 (8): 3478–3486.

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Jinjin Zhou participated in the design of the work, methodology, data interpretation, and analysis for the work, carried out the statistical analysis and drafted the manuscript. Xia Peng participated in the methodology, data interpretation and analysis for the work. Yanhai Ru and Jiayun Xu participated in data interpretation and methodology. All authors read and approved the final manuscript.

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Correspondence to Jinjin Zhou.

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The research related to human use has been complied with all the relevant national regulations, institutional policies and in accordance the tenets of the Helsinki Declaration and has been approved by the Ethics Committee of The First Affiliated Hospital of Henan University of Science and Technology.

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All patients in this study provided their consent for publication.

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Zhou, J., Peng, X., Ru, Y. et al. Circ_0060077 Knockdown Alleviates High-Glucose-Induced Cell Apoptosis, Oxidative Stress, Inflammation and Fibrosis in HK-2 Cells via miR-145-5p/VASN Pathway. Inflammation (2022).

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  • DN
  • HG
  • HK-2
  • circ_0060077
  • miR-145-5p
  • VASN