Abstract
Diabetic nephropathy (DN) is a severe complication of diabetes lethal for end-stage renal disease, with less treatment methodologies and uncertain pathogenesis. In the current study, we determined the role of mesenchymal stem cells (MSCs)-derived extracellular vesicles (EVs) containing microRNA (miR)-15b-5p in DN. After extraction and identification of MSC-derived EVs, mouse podocyte line MPC5 was selected to establish an in vitro high-glucose (HG) cell model, where expression of miR-15b-5p, pyruvate dehydrogenase kinase 4 (PDK4) and VEGFA expression in tissues and cells were determined. The loss- and gain- function assays were conducted to determine the roles of miR-15b-5p, PDK4 and VEGFA. MPC5 cells were then co-cultured with MSC-derived EVs and their biological behaviors were detected by Western blot, CCK-8 assay, and flow cytometry. The binding relationship between miR-15b-5p and PDK43 by dual luciferase reporter gene assay. The expression of miR-15b-5p was downregulated in podocytes under HG environment, but highly expressed in mouse MSCs-derived EVs. EVs-derived miR-15b-5p could protect MPC5 cell apoptosis and inflammation. miR-15b-5p inhibited the expression of PDK4 by directly bound to the 3’UTR region of PDK4 gene. miR-15b-5p inhibits VEGF expression by binding to PDK4. Inhibition of PDK4 decreased VEGFA expression and reduced apoptosis and inflammation. Collectively, miR-15b-5p shuttled by MSC-derived EV can play protective roles in HG-induced mouse podocyte injury, possibly by targeting PDK4 and decreasing the VEGFA expression.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Data will be made available on reasonable request.
References
Atwater JA, Munemitsu SM, Samuel CE (1987) Biosynthesis of reovirus-specified polypeptides. Efficiency of expression of cDNAs of the reovirus S1 and S4 genes in transfected animal cells differs at the level of translation. Virology 159:350–357. https://doi.org/10.1016/0042-6822(87)90473-9
Baek G, Choi H, Kim Y, Lee HC, Choi C (2019) Mesenchymal stem cell-derived extracellular vesicles as therapeutics and as a drug delivery platform. Stem Cells Transl Med 8:880–886. https://doi.org/10.1002/sctm.18-0226
Bai J, Wang Y, Zhu X, Shi J (2019) Eriodictyol inhibits high glucose-induced extracellular matrix accumulation, oxidative stress, and inflammation in human glomerular mesangial cells. Phytother Res 33:2775–2782. https://doi.org/10.1002/ptr.6463
Bertoli G, Cava C, Castiglioni I (2015) MicroRNAs: new biomarkers for diagnosis, prognosis, therapy prediction and therapeutic tools for breast Cancer. Theranostics 5:1122–1143. https://doi.org/10.7150/thno.11543
Bus P, Scharpfenecker M, Van Der Wilk P, Wolterbeek R, Bruijn JA, Baelde HJ (2017) The VEGF-A inhibitor sFLT-1 improves renal function by reducing endothelial activation and inflammation in a mouse model of type 1 diabetes. Diabetologia 60:1813–1821. https://doi.org/10.1007/s00125-017-4322-3
Duan YR, Chen BP, Chen F, Yang SX, Zhu CY, Ma YL, Li Y, Shi J (2019) Exosomal microRNA-16-5p from human urine-derived stem cells ameliorates diabetic nephropathy through protection of podocyte. J Cell Mol Med. https://doi.org/10.1111/jcmm.14558
Fu Y, Wang C, Zhang D, Chu X, Zhang Y, Li J (2019) miR-15b-5p ameliorated high glucose-induced podocyte injury through repressing apoptosis, oxidative stress, and inflammatory responses by targeting Sema3A. J Cell Physiol 234:20869–20878. https://doi.org/10.1002/jcp.28691
Jeong JY, Jeoung NH, Park KG, Lee IK (2012) Transcriptional regulation of pyruvate dehydrogenase kinase. Diabetes Metab J 36:328–335. https://doi.org/10.4093/dmj.2012.36.5.328
Jeoung NH, Harris RA (2010) Role of pyruvate dehydrogenase kinase 4 in regulation of blood glucose levels. Korean Diabetes J 34:274–283. https://doi.org/10.4093/kdj.2010.34.5.274
Johnson SA, Spurney RF (2015) Twenty years after ACEIs and ARBs: emerging treatment strategies for diabetic nephropathy. Am J Physiol Renal Physiol 309:F807–F820. https://doi.org/10.1152/ajprenal.00266.2015
Keshtkar S, Azarpira N, Ghahremani MH (2018) Mesenchymal stem cell-derived extracellular vesicles: novel frontiers in regenerative medicine. Stem Cell Res Ther 9:63. https://doi.org/10.1186/s13287-018-0791-7
Kovacs B, Lumayag S, Cowan C, Xu S (2011) MicroRNAs in early diabetic retinopathy in streptozotocin-induced diabetic rats. Invest Ophthalmol Vis Sci 52:4402–4409. https://doi.org/10.1167/iovs.10-6879
Lee ES, Kwon MH, Kim HM, Kim N, Kim YM, Kim HS, Lee EY, Chung CH (2019a) Dibenzoylmethane ameliorates lipid-induced inflammation and oxidative injury in diabetic nephropathy. J Endocrinol 240:169–179. https://doi.org/10.1530/JOE-18-0206
Lee D, Pagire HS, Pagire SH, Bae EJ, Dighe M, Kim M, Lee KM, Jang YK, Jaladi AK, Jung KY, Yoo EK, Gim HE, Lee S, Choi WI, Chi YI, Song JS, Bae MA, Jeon YH, Lee GH et al (2019b) Correction to discovery of novel pyruvate dehydrogenase kinase 4 inhibitors for potential Oral treatment of metabolic diseases. J Med Chem 62:2849. https://doi.org/10.1021/acs.jmedchem.9b00249
Liu F, Chen Q, Chen F, Wang J, Gong R, He B (2019) The lncRNA ENST00000608794 acts as a competing endogenous RNA to regulate PDK4 expression by sponging miR-15b-5p in dexamethasone induced steatosis. Biochim Biophys Acta Mol Cell Biol Lipids 1864:1449–1457. https://doi.org/10.1016/j.bbalip.2019.07.003
Lu L, Li Y, Wen H, Feng C (2019) Overexpression of miR-15b promotes resistance to Sunitinib in renal cell carcinoma. J Cancer 10:3389–3396. https://doi.org/10.7150/jca.31676
Marycz K, Michalak I, Kocherova I, Maredziak M, Weiss C (2017) The Cladophora glomerata Enriched by Biosorption Process in Cr(III) Improves Viability, and Reduces Oxidative Stress and Apoptosis in Equine Metabolic Syndrome Derived Adipose Mesenchymal Stromal Stem Cells (ASCs) and Their Extracellular Vesicles (MV's). Mar Drugs 15. https://doi.org/10.3390/md15120385
Onions KL, Gamez M, Buckner NR, Baker SL, Betteridge KB, Desideri S, Dallyn BP, Ramnath RD, Neal CR, Farmer LK, Mathieson PW, Gnudi L, Alitalo K, Bates DO, Salmon AHJ, Welsh GI, Satchell SC, Foster RR (2019) VEGFC reduces glomerular albumin permeability and protects against alterations in VEGF receptor expression in diabetic nephropathy. Diabetes 68:172–187. https://doi.org/10.2337/db18-0045
Pin F, Novinger LJ, Huot JR, Harris RA, Couch ME, O'Connell TM, Bonetto A (2019) PDK4 drives metabolic alterations and muscle atrophy in cancer cachexia. FASEB J 33:7778–7790. https://doi.org/10.1096/fj.201802799R
Ramirez HA, Pastar I, Jozic I, Stojadinovic O, Stone RC, Ojeh N, Gil J, Davis SC, Kirsner RS, Tomic-Canic M (2018) Staphylococcus aureus triggers induction of miR-15B-5P to diminish DNA repair and deregulate inflammatory response in diabetic foot ulcers. J Invest Dermatol 138:1187–1196. https://doi.org/10.1016/j.jid.2017.11.038
Rashed LA, Elattar S, Eltablawy N, Ashour H, Mahmoud LM, El-Esawy Y (2018) Mesenchymal stem cells pretreated with melatonin ameliorate kidney functions in a rat model of diabetic nephropathy. Biochem Cell Biol 96:564–571. https://doi.org/10.1139/bcb-2017-0230
Stevens M, Oltean S (2018) Modulation of VEGF-A alternative splicing as a novel treatment in chronic kidney disease. Genes (Basel) 9. https://doi.org/10.3390/genes9020098
Toblli JE, Bevione P, Di Gennaro F, Madalena L, Cao G, Angerosa M (2012) Understanding the mechanisms of proteinuria: therapeutic implications. Int J Nephrol 2012:546039. https://doi.org/10.1155/2012/546039
Tsai YC, Kuo MC, Hung WW, Wu LY, Wu PH, Chang WA, Kuo PL, Hsu YL (2020) High glucose induces Mesangial cell apoptosis through miR-15b-5p and promotes diabetic nephropathy by extracellular vesicle delivery. Mol Ther 28:963–974. https://doi.org/10.1016/j.ymthe.2020.01.014
Weng Y, Shen Y, He Y, Pan X, Xu J, Jiang Y, Zhang Q, Wang S, Kong F, Zhao S, Zhou D, Zhang Y (2018) The miR-15b-5p/PDK4 axis regulates osteosarcoma proliferation through modulation of the Warburg effect. Biochem Biophys Res Commun 503:2749–2757. https://doi.org/10.1016/j.bbrc.2018.08.035
Yanez-Mo M, Siljander PR, Andreu Z, Zavec AB, Borras FE, Buzas EI, Buzas K, Casal E, Cappello F, Carvalho J, Colas E, Cordeiro-da Silva A, Fais S, Falcon-Perez JM, Ghobrial IM, Giebel B, Gimona M, Graner M, Gursel I et al (2015) Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles 4:27066. https://doi.org/10.3402/jev.v4.27066
Yang Y, Hou N, Wang X, Wang L, Chang S, He K, Zhao Z, Zhao X, Song T, Huang C (2015) miR-15b-5p induces endoplasmic reticulum stress and apoptosis in human hepatocellular carcinoma, both in vitro and in vivo, by suppressing Rab1A. Oncotarget 6:16227–16238. https://doi.org/10.18632/oncotarget.3970
Ye EA, Steinle JJ (2015) miR-15b/16 protects primary human retinal microvascular endothelial cells against hyperglycemia-induced increases in tumor necrosis factor alpha and suppressor of cytokine signaling 3. J Neuroinflammation 12:44. https://doi.org/10.1186/s12974-015-0265-0
Zhao C, Li Y, Chen G, Wang F, Shen Z, Zhou R (2017) Overexpression of miR-15b-5p promotes gastric cancer metastasis by regulating PAQR3. Oncol Rep 38:352–358. https://doi.org/10.3892/or.2017.5673
Zhou Q, Zhu Y, Wei X, Zhou J, Chang L, Sui H, Han Y, Piao D, Sha R, Bai Y (2016) MiR-590-5p inhibits colorectal cancer angiogenesis and metastasis by regulating nuclear factor 90/vascular endothelial growth factor a axis. Cell Death Dis 7:e2413. https://doi.org/10.1038/cddis.2016.306
Zhu X, Shi J, Li H (2018) Liquiritigenin attenuates high glucose-induced mesangial matrix accumulation, oxidative stress, and inflammation by suppression of the NF-kappaB and NLRP3 inflammasome pathways. Biomed Pharmacother 106:976–982. https://doi.org/10.1016/j.biopha.2018.07.045
Zhuo HC, Wang BE (1985) Preliminary clinical study of the effect of complex traditional Chinese medicine pills (848) on chronic hepatitis B virus carriers. Zhong Xi Yi Jie He Za Zhi 5:606–608, 579
Funding
This study was supported by the Natural Science Foundation of Shandong Province (No. ZR2015HL029).
Author information
Authors and Affiliations
Contributions
Tiantian Zhao, Dongdong Zhang and Minglong Li drafted manuscript, conceived and designed research and interpreted results of experiments; Qingsong Jin, Yaqin Teng and Liangyan Lin analyzed data and prepared figures; Lili Kong, Xiaoyan Yao and Yongjun Jin performed experiments and edited and revised manuscript. All authors have read and approved the final submitted manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
ESM 1
(DOC 33 kb)
Rights and permissions
About this article
Cite this article
Zhao, T., Jin, Q., Kong, L. et al. microRNA-15b-5p shuttled by mesenchymal stem cell-derived extracellular vesicles protects podocytes from diabetic nephropathy via downregulation of VEGF/PDK4 axis. J Bioenerg Biomembr 54, 17–30 (2022). https://doi.org/10.1007/s10863-021-09919-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10863-021-09919-y