MicroRNA-5010-5p Ameliorates High-glucose Induced Inflammation in Renal Tubular Epithelial Cells by Modulating the Expression of PPP2R2D

Overview

Journal BMJ Open Diabetes Res Care

Specialty Endocrinology

Date 2024 Mar 5

PMID 38442987

Authors

Sunghee Choi

Mithun Kumer Sarker

Mi Ra Yu

Haekyung Lee

Soon Hyo Kwon

Jin Seok Jeon

Hyunjin Noh

Hyoungnae Kim

Affiliations

Soon will be listed here.

Abstract

Introduction: We previously reported the significant upregulation of eight circulating exosomal microRNAs (miRNAs) in patients with diabetic kidney disease (DKD). However, their specific roles and molecular mechanisms in the kidney remain unknown. Among the eight miRNAs, we evaluated the effects of miR-5010-5p on renal tubular epithelial cells under diabetic conditions in this study.

Research Design And Methods: We transfected the renal tubular epithelial cell line, HK-2, with an miR-5010-5p mimic using recombinant plasmids. The target gene of hsa-miR-5010-5p was identified using a dual-luciferase assay. Cell viability was assessed via the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay. Moreover, mRNA and protein expression levels were determined via real-time PCR and western blotting, respectively.

Results: High glucose levels did not significantly affect the intracellular expression of miR-5010-5p in HK-2 cells. Transfection of the miR-5010-5p mimic caused no change in cell viability. However, miR-5010-5p-transfected HK-2 cells exhibited significantly decreased expression levels of inflammatory cytokines, such as the monocyte chemoattractant protein-1, interleukin-1β, and tumor necrosis factor-ɑ, under high-glucose conditions. These changes were accompanied by the restored expression of phosphorylated AMP-activated protein kinase (AMPK) and decreased phosphorylation of nuclear factor-kappa B. Dual-luciferase assay revealed that miR-5010-5p targeted the gene, protein phosphatase 2 regulatory subunit B delta (PPP2R2D), a subunit of protein phosphatase 2A, which modulates AMPK phosphorylation.

Conclusions: Our findings suggest that increased miR-5010-5p expression reduces high glucose-induced inflammatory responses in renal tubular epithelial cells via the regulation of the target gene, PPP2R2D, which modulates AMPK phosphorylation. Therefore, miR-5010-5p may be a promising therapeutic target for DKD.

References

Du Y, Kowluru A, Kern T . PP2A contributes to endothelial death in high glucose: inhibition by benfotiamine. Am J Physiol Regul Integr Comp Physiol. 2010; 299(6):R1610-7. PMC: 3774204. DOI: 10.1152/ajpregu.00676.2009. View

Conserva F, Barozzino M, Pesce F, Divella C, Oranger A, Papale M . Urinary miRNA-27b-3p and miRNA-1228-3p correlate with the progression of Kidney Fibrosis in Diabetic Nephropathy. Sci Rep. 2019; 9(1):11357. PMC: 6684817. DOI: 10.1038/s41598-019-47778-1. View

Kovesdy C . Epidemiology of chronic kidney disease: an update 2022. Kidney Int Suppl (2011). 2022; 12(1):7-11. PMC: 9073222. DOI: 10.1016/j.kisu.2021.11.003. View

Kato M, Zhang J, Wang M, Lanting L, Yuan H, Rossi J . MicroRNA-192 in diabetic kidney glomeruli and its function in TGF-beta-induced collagen expression via inhibition of E-box repressors. Proc Natl Acad Sci U S A. 2007; 104(9):3432-7. PMC: 1805579. DOI: 10.1073/pnas.0611192104. View

Salminen A, Kaarniranta K, Kauppinen A . Age-related changes in AMPK activation: Role for AMPK phosphatases and inhibitory phosphorylation by upstream signaling pathways. Ageing Res Rev. 2016; 28:15-26. DOI: 10.1016/j.arr.2016.04.003. View

Choi S, Lim J, Kim M, Kim E, Kim Y, Choi B . Adiponectin receptor agonist AdipoRon decreased ceramide, and lipotoxicity, and ameliorated diabetic nephropathy. Metabolism. 2018; 85:348-360. DOI: 10.1016/j.metabol.2018.02.004. View

Zhou P, Shaffer D, Alvarez Arias D, Nakazaki Y, Pos W, Torres A . In vivo discovery of immunotherapy targets in the tumour microenvironment. Nature. 2014; 506(7486):52-7. PMC: 4052214. DOI: 10.1038/nature12988. View

Zhong Y, Lee K, Deng Y, Ma Y, Chen Y, Li X . Arctigenin attenuates diabetic kidney disease through the activation of PP2A in podocytes. Nat Commun. 2019; 10(1):4523. PMC: 6778111. DOI: 10.1038/s41467-019-12433-w. View

Tang J, Yao D, Yan H, Chen X, Wang L, Zhan H . The Role of MicroRNAs in the Pathogenesis of Diabetic Nephropathy. Int J Endocrinol. 2019; 2019:8719060. PMC: 6914872. DOI: 10.1155/2019/8719060. View

10.

Wang B, Yao K, Huuskes B, Shen H, Zhuang J, Godson C . Mesenchymal Stem Cells Deliver Exogenous MicroRNA-let7c via Exosomes to Attenuate Renal Fibrosis. Mol Ther. 2016; 24(7):1290-301. PMC: 5088767. DOI: 10.1038/mt.2016.90. View

11.

Joseph B, Liu H, Francisco J, Pandya D, Donigan M, Gallo-Ebert C . Inhibition of AMP Kinase by the Protein Phosphatase 2A Heterotrimer, PP2APpp2r2d. J Biol Chem. 2015; 290(17):10588-98. PMC: 4409226. DOI: 10.1074/jbc.M114.626259. View

12.

Ma H, Guo X, Cui S, Wu Y, Zhang Y, Shen X . Dephosphorylation of AMP-activated protein kinase exacerbates ischemia/reperfusion-induced acute kidney injury via mitochondrial dysfunction. Kidney Int. 2021; 101(2):315-330. DOI: 10.1016/j.kint.2021.10.028. View

13.

Kato M, Natarajan R . MicroRNAs in diabetic nephropathy: functions, biomarkers, and therapeutic targets. Ann N Y Acad Sci. 2015; 1353:72-88. PMC: 4607544. DOI: 10.1111/nyas.12758. View

14.

Barr F, Elliott P, Gruneberg U . Protein phosphatases and the regulation of mitosis. J Cell Sci. 2011; 124(Pt 14):2323-34. DOI: 10.1242/jcs.087106. View

15.

. Impact of diabetes on the effects of sodium glucose co-transporter-2 inhibitors on kidney outcomes: collaborative meta-analysis of large placebo-controlled trials. Lancet. 2022; 400(10365):1788-1801. PMC: 7613836. DOI: 10.1016/S0140-6736(22)02074-8. View

16.

Hashemi Sheikhshabani S, Amini-Farsani Z, Modarres P, Amini-Farsani Z, Khazaei Feyzabad S, Shaygan N . In silico identification of potential miRNAs -mRNA inflammatory networks implicated in the pathogenesis of COVID-19. Hum Gene (Amst). 2023; 36:201172. PMC: 10085880. DOI: 10.1016/j.humgen.2023.201172. View

17.

Pan W, Sharabi A, Ferretti A, Zhang Y, Burbano C, Yoshida N . PPP2R2D suppresses IL-2 production and Treg function. JCI Insight. 2020; 5(19). PMC: 7566706. DOI: 10.1172/jci.insight.138215. View

18.

Kim H, Bae Y, Jeon J, Noh H, Park H, Byun D . The circulating exosomal microRNAs related to albuminuria in patients with diabetic nephropathy. J Transl Med. 2019; 17(1):236. PMC: 6647278. DOI: 10.1186/s12967-019-1983-3. View

19.

Liu S, Wu W, Liao J, Tang F, Gao G, Peng J . MicroRNA-21: A Critical Pathogenic Factor of Diabetic Nephropathy. Front Endocrinol (Lausanne). 2022; 13:895010. PMC: 9294636. DOI: 10.3389/fendo.2022.895010. View

20.

Liu J, Li W, Li J, Liu F, Zhou L . Screening key long non-coding RNAs in early-stage colon adenocarcinoma by RNA-sequencing. Epigenomics. 2018; 10(9):1215-1228. DOI: 10.2217/epi-2017-0155. View