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CircRNA circ-ITCH improves renal inflammation and fibrosis in streptozotocin-induced diabetic mice by regulating the miR-33a-5p/SIRT6 axis

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Abstract

Diabetic nephropathy (DN) seriously affects the people's life and health in China. This study aimed to investigate the effect of circRNA circ-ITCH on improving DN by regulating the miR-33a-5p/SIRT6 axis and the possible mechanism of action. High glucose (HG)-induced rat mesangial cells (RMCs) were used to simulate the DN in vitro. Reverse transcription-quantitative PCR (RT-qPCR) and western blot analysis were conducted to detect the gene or protein expression. Cell Counting Kit-8 (CCK-8) and wound healing assays were performed to estimate the cell viability and migration capability. Immunofluorescence and enzyme-linked immunosorbent assay (ELISA) were used to detect the α-Smooth Muscle Actin (α-SMA) expression and levels of inflammatory factors. The potential associations between circ-ITCH and miR-33a-5p, miR-33a-5p and SIRT6 in RMCs were measured via dual-luciferase reporter assay. Streptozotocin (STZ) was used to induce the diabetic mice. Blood glucose and serum insulin of mice were determined by corresponding kits, and blood urea nitrogen (BUN) and serum creatinine (SCr) were measured using an automatic biochemical analyzer. Hematoxylin and eosin (H&E) staining and periodic acid-Schiff (PAS) staining were applied to observe the degree of pathological injury and fibrosis of renal tissues. The results of the present study revealed that circ-ITCH expression was obviously decreased in HG-induced RMCs. In addition, circ-ITCH overexpression inhibited the viability, migration, fibrosis and inflammatory response of HG-induced RMCs. Further experiments confirmed that miR-33a-5p may be a direct target of circ-ITCH and SIRT6 may be a direct target of miR-33a-5p. Notably, the miR-33a-5p mimic or shRNA-SIRT6 were discovered to reverse the inhibitory effects of circ-ITCH on the proliferation, migration, fibrosis and inflammatory response of HG-induced RMCs. Furthermore, circ-ITCH overexpression ameliorated renal inflammation and fibrosis in STZ-induced diabetic mice. In conclusion, circ-ITCH alleviated renal inflammation and fibrosis in STZ-induced diabetic mice by regulating the miR-33a-5p/SIRT6 axis.Author names: Please confirm if the author names are presented accurately and in the correct sequence (given name, middle name/initial, family name). Author 1 Given name: [ChunYang] Last name [Xu]. Author 2 Given name: [DingBo] Last name [Xu]. Author 3 Given name: [YongHua] Last name [Liu]. Author 4 Given name: [Juanjuan] Last name [Jiang]. Also, kindly confirm the details in the metadata are correct.ok

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References

  1. Kato M, Natarajan R. Epigenetics and epigenomics in diabetic kidney disease and metabolic memory. Nat Rev Nephrol. 2019;15:327–45.

    Article  Google Scholar 

  2. Lan HY. Transforming growth factor-β/Smad signalling in diabetic nephropathy. Clin Exp Pharmacol Physiol. 2012;39:731–8.

    Article  CAS  Google Scholar 

  3. Kashgarian M, Fogo AB. Diagnostic atlas of renal pathology. 3rd ed. Amsterdam: Elsevier; 2017.

    Google Scholar 

  4. Memczak S, Jens M, Elefsinioti A, Torti F, Krueger J, Rybak A, et al. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature. 2013;495:333–8.

    Article  CAS  Google Scholar 

  5. Kristensen LS, Andersen MS, Stagsted LVW, Ebbesen KK, Hansen TB, Kjems J. The biogenesis, biology and characterization of circular RNAs. Nat Rev Genet. 2019;20:675–91.

    Article  CAS  Google Scholar 

  6. Meng S, Zhou H, Feng Z, Xu Z, Tang Y, Li P, et al. CircRNA: functions and properties of a novel potential biomarker for cancer. Mol Cancer. 2017;16:94.

    Article  Google Scholar 

  7. Yin Y, Long J, He Q, Li Y, Liao Y, He P, et al. Emerging roles of circRNA in formation and progression of cancer. J Cancer. 2019;10:5015–21.

    Article  CAS  Google Scholar 

  8. Liu Y, Yang Y, Wang Z, Fu X, Chu XM, Li Y, et al. Insights into the regulatory role of circRNA in angiogenesis and clinical implications. Atherosclerosis. 2020;298:14–26.

    Article  CAS  Google Scholar 

  9. Tian J, Fu Y, Li Q, Xu Y, Xi X, Zheng Y, et al. Differential expression and bioinformatics analysis of CircRNA in PDGF-BB-induced vascular smooth muscle cells. Front Genet. 2020;11:530.

    Article  CAS  Google Scholar 

  10. Yan Q, He X, Kuang G, Ou C. CircRNA cPWWP2A: an emerging player in diabetes mellitus. J Cell Commun Signal. 2020;14:351–3.

    Article  Google Scholar 

  11. Yang F, Chen Y, Xue Z, Lv Y, Shen L, Li K, et al. High-throughput sequencing and exploration of the lncRNA-circRNA-miRNA-mRNA network in Type 2 diabetes mellitus. Biomed Res Int. 2020;2020:8162524.

    PubMed  PubMed Central  Google Scholar 

  12. Peng F, Gong W, Li S, Yin B, Zhao C, Liu W, et al. circRNA_010383 acts as a sponge for miR-135a and its downregulated expression contributes to renal fibrosis in diabetic nephropathy. Diabetes. 2020. https://doi.org/10.2337/db200203.

    Article  PubMed  Google Scholar 

  13. Zhou B, Yu JW. A novel identified circular RNA, circRNA_010567, promotes myocardial fibrosis via suppressing miR-141 by targeting TGF-β1. Biochem Biophys Res Commun. 2017;487:769–75.

    Article  CAS  Google Scholar 

  14. Hu W, Han Q, Zhao L, Wang L. Circular RNA circRNA_15698 aggravates the extracellular matrix of diabetic nephropathy mesangial cells via miR-185/TGF-β1. J Cell Physiol. 2019;234:1469–76.

    Article  CAS  Google Scholar 

  15. Zhou L, Li FF, Wang SM. Circ-ITCH restrains the expression of MMP-2, MMP-9 and TNF-α in diabetic retinopathy by inhibiting miR-22. Exp Mol Pathol. 2021;118:104594.

    Article  CAS  Google Scholar 

  16. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001;25:402–8.

    Article  CAS  Google Scholar 

  17. Su HC, Hung LM, Chen JK. Resveratrol, a red wine antioxidant, possesses an insulin-like effect in streptozotocin-induced diabetic rats. Am J Physiol Endocrinol Metab. 2006;290:E1339–46.

    Article  CAS  Google Scholar 

  18. Yamashita H, Nagai Y, Takamura T, Nohara E, Kobayashi K. Thiazolidinedione derivatives ameliorate albuminuria in streptozotocin-induced diabetic spontaneous hypertensive rat. Metabolism. 2002;51:403–8.

    Article  CAS  Google Scholar 

  19. Zhang M, Lv XY, Li J, Xu ZG, Chen L. The characterization of high-fat diet and multiple low-dose streptozotocin induced type 2 diabetes rat model. Exp Diabetes Res. 2008;2008:704045.

    Article  Google Scholar 

  20. Ogurtsova K, da Rocha Fernandes JD, Huang Y, Linnenkamp U, Guariguata L, Cho NH, et al. IDF Diabetes Atlas: Global estimates for the prevalence of diabetes for 2015 and 2040. Diabetes Res Clin Pract. 2017;128:40–50.

    Article  CAS  Google Scholar 

  21. Cho NH, Shaw JE, Karuranga S, Huang Y, da Rocha Fernandes JD, Ohlrogge AW, et al. IDF Diabetes Atlas: global estimates of diabetes prevalence for 2017 and projections for 2045. Diabetes Res Clin Pract. 2018;138:271–81.

    Article  CAS  Google Scholar 

  22. Cho NH. Q&A: five questions on the 2015 IDF diabetes atlas. Diabetes Res Clin Pract. 2016;115:157–9.

    Article  Google Scholar 

  23. Zhang L, Long J, Jiang W, Shi Y, He X, Zhou Z, et al. Trends in chronic kidney disease in China. N Engl J Med. 2016;375:905–6.

    Article  Google Scholar 

  24. Wang ST, Liu LB, Li XM, Wang YF, Xie PJ, Li Q, et al. Circ-ITCH regulates triple-negative breast cancer progression through the Wnt/β-catenin pathway. Neoplasma. 2019;66:232–9.

    Article  CAS  Google Scholar 

  25. Ren C, Liu J, Zheng B, Yan P, Sun Y, Yue B. The circular RNA circ-ITCH acts as a tumour suppressor in osteosarcoma via regulating miR-22. Artif Cells Nanomed Biotechnol. 2019;47:3359–67.

    Article  CAS  Google Scholar 

  26. Hu J, Wang L, Chen J, Gao H, Zhao W, Huang Y, et al. The circular RNA circ-ITCH suppresses ovarian carcinoma progression through targeting miR-145/RASA1 signaling. Biochem Biophys Res Commun. 2018;505:222–8.

    Article  CAS  Google Scholar 

  27. Guo W, Zhang J, Zhang D, Cao S, Li G, Zhang S, et al. Polymorphisms and expression pattern of circular RNA circ-ITCH contributes to the carcinogenesis of hepatocellular carcinoma. Oncotarget. 2017;8:48169–77.

    Article  Google Scholar 

  28. Feng Y, Qu X, Chen Y, Feng Q, Zhang Y, Hu J, et al. MicroRNA-33a-5p sponges to inhibit pancreatic β-cell function in gestational diabetes mellitus LncRNA DANCR. Reprou Biol Endocrinol. 2020;18:61.

    Article  CAS  Google Scholar 

  29. Coskun ZM, Beydogan AB, Bolkent S. Changes in the expression levels of CB1 and GLP-1R mRNAs and microRNAs 33a and 122 in the liver of type 2 diabetic rats treated with ghrelin. J Biochem Mol Toxicol. 2019;33:e22388.

    Article  CAS  Google Scholar 

  30. Xu BH, Sheng J, You YK, Huang XR, Ma RCW, Wang Q, et al. Deletion of Smad3 prevents renal fibrosis and inflammation in type 2 diabetic nephropathy. Metabolism. 2020;103:154013.

    Article  CAS  Google Scholar 

  31. Huang H, Ni H, Ma K, Zou J. ANGPTL2 regulates autophagy through the MEK/ERK/Nrf-1 pathway and affects the progression of renal fibrosis in diabetic nephropathy. Am J Transl Res. 2019;11:5472–86.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Yi H, Peng R, Zhang LY, Sun Y, Peng HM, Liu HD, et al. LincRNA-Gm4419 knockdown ameliorates NF-κB/NLRP3 inflammasome-mediated inflammation in diabetic nephropathy. Cell Death Dis. 2017;8:e2583.

    Article  CAS  Google Scholar 

  33. Wada J, Makino H. Inflammation and the pathogenesis of diabetic nephropathy. Clin Sci. 2013;124:139–52.

    Article  CAS  Google Scholar 

  34. Chen Z, Ding HS, Guo X, Shen JJ, Fan D, Huang Y, et al. MiR-33 promotes myocardial fibrosis by inhibiting MMP16 and stimulating p38 MAPK signaling. Oncotarget. 2018;9:22047–57.

    Article  Google Scholar 

  35. Erhartova D, Cahova M, Dankova H, Heczkova M, Mikova I, Sticova E, et al. Serum miR-33a is associated with steatosis and inflammation in patients with non-alcoholic fatty liver disease after liver transplantation. PLoS ONE. 2019;14:e0224820.

    Article  CAS  Google Scholar 

  36. Vega-Badillo J, Gutiérrez-Vidal R, Hernández-Pérez HA, Villamil-Ramírez H, León-Mimila P, Sánchez-Muñoz F, et al. Hepatic miR-33a/miR-144 and their target gene ABCA1 are associated with steatohepatitis in morbidly obese subjects. Liver Int. 2016;36:1383–91.

    Article  CAS  Google Scholar 

  37. Ji L, Chen Y, Wang H, Zhang W, He L, Wu J, et al. Overexpression of Sirt6 promotes M2 macrophage transformation, alleviating renal injury in diabetic nephropathy. Int J Oncol. 2019;55:103–15.

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Fan Y, Yang Q, Yang Y, Gao Z, Ma Y, Zhang L, et al. Sirt6 suppresses high glucose-induced mitochondrial dysfunction and apoptosis in podocytes through AMPK activation. Int J Biol Sci. 2019;15:701–13.

    Article  CAS  Google Scholar 

  39. Muraoka H, Hasegawa K, Sakamaki Y, Minakuchi H, Kawaguchi T, Yasuda I, et al. Role of nampt-Sirt6 axis in renal proximal tubules in extracellular matrix deposition in diabetic nephropathy. Cell Rep. 2019;27:199-212.e5.

    Article  CAS  Google Scholar 

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Funding

National Natural Science Foundation of China (Project No. 81760153); Key R & D Program of Jiangxi Province (Project No. 20181BBG70014); Key project of Key R & D Program of Jiangxi Province (Project No. 20171ACH80002).

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Author notes

  1. Jiang Liu and Peng Duan contributed equally.

Authors and Affiliations

  1. Department of Endocrinology and Metabolism, The Third Hospital of Nanchang, Nanchang, 330009, Jiangxi province, China

    Jiang Liu, Peng Duan, DingBo Xu & YongHua Liu

  2. School of Medicine, Southeast University, Nanjing, 211189, Jiangsu province, China

    Jiang Liu

  3. Department of Cardiovascular Medicine, Yancheng No.1 People’s Hospital, Yancheng, 224006, Jiangsu province, China

    ChunYang Xu

  4. Department of Nephrology, Jiangsu Province Geriatric Hospital, Geriatric Hospital of Nanjing Medical University, Jiangsu Province Official Hospital, 65 Jiangsu road, Nanjing, 210024, Jiangsu province, China

    Juanjuan Jiang

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  1. Jiang Liu
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  2. Peng Duan
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  3. ChunYang Xu
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  4. DingBo Xu
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  5. YongHua Liu
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Contributions

JL: project development, literature search, data analysis and collection, manuscript writing. PD: project development, literature search, data analysis and collection, manuscript writing. CX: project development, data collection. DX: project development, data collection. YL: project development, data collection. JJ: project development, manuscript editing.

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Correspondence to Juanjuan Jiang.

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The authors declare they have no competing interests.

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Animal experiments were approved by the Ethics Committee of Chinese PLA General Hospital.

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Responsible Editor: John Di Battista.

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Liu, J., Duan, P., Xu, C. et al. CircRNA circ-ITCH improves renal inflammation and fibrosis in streptozotocin-induced diabetic mice by regulating the miR-33a-5p/SIRT6 axis. Inflamm. Res. 70, 835–846 (2021). https://doi.org/10.1007/s00011-021-01485-8

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  • DOI: https://doi.org/10.1007/s00011-021-01485-8

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