MicroRNA‑145‑5p attenuates high glucose‑induced apoptosis by targeting the Notch signaling pathway in podocytes

Wei,Bing; Liu,Yi‑Song; Guan,Hai‑Xia

doi:10.3892/etm.2020.8427

MicroRNA‑145‑5p attenuates high glucose‑induced apoptosis by targeting the Notch signaling pathway in podocytes

Authors:
- Bing Wei
- Yi‑Song Liu
- Hai‑Xia Guan
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Affiliations: Department of Endocrinology, Daqing Oil Field General Hospital, Daqing, Heilongjiang 163411, P.R. China, Dental Department, Daqing Oil Field General Hospital, Daqing, Heilongjiang 163411, P.R. China, Department of Endocrinology, The First Hospital of China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: January 7, 2020 https://doi.org/10.3892/etm.2020.8427
Pages: 1915-1924

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Abstract

MicroRNAs (miRNAs/miRs) are considered to serve essential roles in podocyte apoptosis, and to be critical in the development of diabetic nephropathy (DN). Activation of the Notch signaling pathway has been demonstrated to serve an important role in DN development; however, its regulatory mechanisms are not fully understood. The present study used a high glucose (HG)‑induced in vitro apoptosis model using mouse podocytes. Expression levels of miR‑145‑5p and its target, Notch1, and other key factors involved in the apoptosis signaling pathway were detected and measured by reverse transcription‑quantitative PCR and western blotting. A luciferase reporter assay was performed to elucidate the miRNA‑target interactions. The functions of miR‑145‑5p in apoptosis were detected using flow cytometry and TUNEL staining. The present study demonstrated that in HG conditions, miR‑145‑5p overexpression inhibited Notch1, Notch intracellular domain, Hes1 and Hey1 expression at the mRNA and protein levels. Notch1 was identified as a direct target of miR‑145‑5p. Furthermore, cleaved caspase‑3, Bcl‑2 and Bax levels were reduced significantly by miR‑145‑5p overexpression. These results indicate that miR‑145‑5p overexpression inhibited the Notch signaling pathway and podocyte lesions induced by HG. In conclusion, the results of the present study suggested that miR‑145‑5p may be a regulator of DN. Additionally, miR‑145‑5p inhibited HG‑induced apoptosis by directly targeting Notch1 and dysregulating apoptotic factors, including cleaved caspase‑3, Bcl‑2 and Bax. The results of the present study provided evidence that miR‑145‑5p may offer a novel approach for the treatment of DN.

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1	Tong L and Adler SG: Diabetic kidney disease. Clin J Am Soc Nephrol. 13:335–338. 2018. View Article : Google Scholar : PubMed/NCBI
2	Ying Q and Wu G: Molecular mechanisms involved in podocyte EMT and concomitant diabetic kidney diseases: An update. Ren Fail. 39:474–483. 2017. View Article : Google Scholar : PubMed/NCBI
3	Asanuma K: The role of podocyte injury in chronic kidney disease. Nihon Rinsho Meneki Gakkai Kaishi. 38:26–36. 2015.(In Japanese). View Article : Google Scholar : PubMed/NCBI
4	Brosius FC and Coward RJ: Podocytes, signaling pathways and vascular factors in diabetic kidney disease. Adv Chronic Kidney Dis. 21:304–310. 2014. View Article : Google Scholar : PubMed/NCBI
5	Armelloni S, Corbelli A, Giardino L, Li M, Ikehata M, Mattinzoli D, Messa P, Pignatari C, Watanabe S and Rastaldi MP: Podocytes: Recent biomolecular developments. Biomol Concepts. 5:319–330. 2014. View Article : Google Scholar : PubMed/NCBI
6	Tetreault N and De Guire V: miRNAs: Their discovery, biogenesis and mechanism of action. Clin Biochem. 46:842–845. 2013. View Article : Google Scholar : PubMed/NCBI
7	Duarte FV, Palmeira CM and Rolo AP: The role of microRNAs in mitochondria: Small players acting wide. Genes (Basel). 5:865–886. 2014. View Article : Google Scholar : PubMed/NCBI
8	Ambros V: The functions of animal microRNAs. Nature. 431:350–354. 2004. View Article : Google Scholar : PubMed/NCBI
9	Li T and Cho WC: MicroRNAs: Mechanisms, functions and progress. Genomics Proteomics Bioinformatics. 10:237–238. 2012. View Article : Google Scholar : PubMed/NCBI
10	Bartel DP: MicroRNAs: Target recognition and regulatory functions. Cell. 136:215–233. 2009. View Article : Google Scholar : PubMed/NCBI
11	Graves P and Zeng Y: Biogenesis of mammalian microRNAs: A global view. Genomics Proteomics Bioinformatics. 10:239–245. 2012. View Article : Google Scholar : PubMed/NCBI
12	Fu Y, Wang C, Zhang D, Chu X, Zhang Y and Li J: 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. 2019. View Article : Google Scholar : PubMed/NCBI
13	Ma J, Li YT, Zhang SX, Fu SZ and Ye XZ: miR-590-3p attenuates acute kidney injury by inhibiting tumor necrosis factor receptor-associated factor 6 in septic mice. Inflammation. 42:637–649. 2019. View Article : Google Scholar : PubMed/NCBI
14	Li H, Zhu X, Zhang J and Shi J: MicroRNA-25 inhibits high glucose-induced apoptosis in renal tubular epithelial cells via PTEN/AKT pathway. Biomed Pharmacother. 96:471–479. 2017. View Article : Google Scholar : PubMed/NCBI
15	Peng J, Wu Y, Deng Z, Zhou Y, Song T, Yang Y, Zhang X, Xu T, Xia M, Cai A, et al: miR-377 promotes white adipose tissue inflammation and decreases insulin sensitivity in obesity via suppression of sirtuin-1 (SIRT1). Oncotarget. 8:70550–70563. 2017. View Article : Google Scholar : PubMed/NCBI
16	Barutta F, Tricarico M, Corbelli A, Annaratone L, Pinach S, Grimaldi S, Bruno G, Cimino D, Taverna D, Deregibus MC, et al: Urinary exosomal microRNAs in incipient diabetic nephropathy. PLoS One. 8:e737982013. View Article : Google Scholar : PubMed/NCBI
17	Yamamoto S, Schulze KL and Bellen HJ: Introduction to Notch signaling. Methods Mol Biol. 1187:1–14. 2014. View Article : Google Scholar : PubMed/NCBI
18	Hori K, Sen A and Artavanis-Tsakonas S: Notch signaling at a glance. J Cell Sci. 126:2135–2140. 2013. View Article : Google Scholar : PubMed/NCBI
19	Penton AL, Leonard LD and Spinner NB: Notch signaling in human development and disease. Semin Cell Dev Biol. 23:450–457. 2012. View Article : Google Scholar : PubMed/NCBI
20	Braune EB and Lendahl U: Notch-a goldilocks signaling pathway in disease and cancer therapy. Discov Med. 21:189–196. 2016.PubMed/NCBI
21	Voelkel JE, Harvey JA, Adams JS, Lassiter RN and Stark MR: FGF and Notch signaling in sensory neuron formation: A multifactorial approach to understanding signaling pathway hierarchy. Mech Dev. 134:55–66. 2014. View Article : Google Scholar : PubMed/NCBI
22	Saleem MA, O'Hare MJ, Reiser J, Coward RJ, Inward CD, Farren T, Xing CY, Ni L, Mathieson PW and Mundel P: A conditionally immortalized human podocyte cell line demonstrating nephrin and podocin expression. J Am Soc Nephrol. 13:630–638. 2002.PubMed/NCBI
23	Shepard BD, Natarajan N, Protzko RJ, Acres OW and Pluznick JL: A cleavable N-terminal signal peptide promotes widespread olfactory receptor surface expression in HEK293T cells. PLoS One. 8:e687582013. View Article : Google Scholar : PubMed/NCBI
24	Loo DT: In situ detection of apoptosis by the TUNEL assay: An overview of techniques. Methods Mol Biol. 682:3–13. 2011. View Article : Google Scholar : PubMed/NCBI
25	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
26	Shan H, Zhang Y, Lu Y, Zhang Y, Pan Z, Cai B, Wang N, Li X, Feng T, Hong Y and Yang B: Downregulation of miR-133 and miR-590 contributes to nicotine-induced atrial remodelling in canines. Cardiovasc Res. 83:465–472. 2009. View Article : Google Scholar : PubMed/NCBI
27	Pockley AG, Foulds GA, Oughton JA, Kerkvliet NI and Multhoff G: Immune cell phenotyping using flow cytometry. Curr Protoc Toxicol. 66:18.8.1–34. 2015. View Article : Google Scholar
28	Agarwal V, Bell GW, Nam J and Bartel DP: Predicting effective microRNA target sites in mammalian mRNAs. eLife. 4:e050052015. View Article : Google Scholar
29	Liu W and Wang X: Prediction of functional microRNA targets by integrative modeling of microRNA binding and target expression data. Genome Biol. 20:182019. View Article : Google Scholar : PubMed/NCBI
30	Zheng R, Pan L, Gao J, Ye X, Chen L, Zhang X, Tang W and Zheng W: Prognostic value of miR-106b expression in breast cancer patients. J Surg Res. 195:158–165. 2015. View Article : Google Scholar : PubMed/NCBI
31	Liu ZH, Dai XM and Du B: Hes1: A key role in stemness, metastasis and multidrug resistance. Cancer Biol Ther. 16:353–359. 2015. View Article : Google Scholar : PubMed/NCBI
32	Lopez-Mateo I, Arruabarrena-Aristorena A, Artaza-Irigaray C, Lopez JA, Calvo E and Belandia B: HEY1 functions are regulated by its phosphorylation at Ser-68. Biosci Rep. 36(pii): e003432016. View Article : Google Scholar : PubMed/NCBI
33	Choudhary GS, Al-Harbi S and Almasan A: Caspase-3 activation is a critical determinant of genotoxic stress-induced apoptosis. Methods Mol Biol. 1219:1–9. 2015. View Article : Google Scholar : PubMed/NCBI
34	Laulier C and Lopez BS: The secret life of Bcl-2: Apoptosis- independent inhibition of DNA repair by Bcl-2 family members. Mutat Res. 751:247–257. 2012. View Article : Google Scholar : PubMed/NCBI
35	Zhu S, Li T, Tan J, Yan X, Zhang D, Zheng C, Chen Y, Xiang Z and Cui H: Bax is essential for death receptor-mediated apoptosis in human colon cancer cells. Cancer Biother Radiopharm. 27:577–581. 2012. View Article : Google Scholar : PubMed/NCBI
36	Kim D, Lim S, Park M, Choi J, Kim J, Han H, Yoon K, Kim K, Lim J and Park S: Ubiquitination-dependent CARM1 degradation facilitates Notch1-mediated podocyte apoptosis in diabetic nephropathy. Cell Signal. 26:1774–1782. 2014. View Article : Google Scholar : PubMed/NCBI
37	Matoba K, Kawanami D, Nagai Y, Takeda Y, Akamine T, Ishizawa S, Kanazawa Y, Yokota T and Utsunomiya K: Rho-kinase blockade attenuates podocyte apoptosis by inhibiting the notch signaling pathway in diabetic nephropathy. Int J Mol Sci. 18(pii): E17952017. View Article : Google Scholar : PubMed/NCBI
38	Gao F, Yao M, Cao Y, Liu S, Liu Q and Duan H: Valsartan ameliorates podocyte loss in diabetic mice through the Notch pathway. Int J Mol Med. 37:1328–1336. 2016. View Article : Google Scholar : PubMed/NCBI
39	Yamamoto S, Schulze KL and Bellen HJ: Introduction to Notch signaling. Notch Signaling: Methods and Protocols. Bellen HJ and Yamamoto S: Springer; New York, NY: pp. 1–14. 2014
40	Sun J, Zhao F, Zhang W, Lv J, Lv J and Yin A: BMSCs and miR-124a ameliorated diabetic nephropathy via inhibiting notch signalling pathway. J Cell Mol Med. 22:4840–4855. 2018. View Article : Google Scholar : PubMed/NCBI
41	Zhang X, Song S and Luo H: Regulation of podocyte lesions in diabetic nephropathy via miR-34a in the Notch signaling pathway. Medicine (Baltimore). 95:e50502016. View Article : Google Scholar : PubMed/NCBI
42	Liu XD, Zhang LY, Zhu TC, Zhang RF, Wang SL and Bao Y: Overexpression of miR-34c inhibits high glucose-induced apoptosis in podocytes by targeting Notch signaling pathways. Int J Clin Exp Pathol. 8:4525–4534. 2015.PubMed/NCBI
43	Chen M, Zhang Y, Li W and Yang J: MicroRNA-145 alleviates high glucose-induced proliferation and migration of vascular smooth muscle cells through targeting ROCK1. Biomed Pharmacother. 99:81–86. 2018. View Article : Google Scholar : PubMed/NCBI
44	Hui Y and Yin Y: MicroRNA-145 attenuates high glucose-induced oxidative stress and inflammation in retinal endothelial cells through regulating TLR4/NF-κB signaling. Life Sci. 207:212–218. 2018. View Article : Google Scholar : PubMed/NCBI
45	Lim JH, Youn DY, Yoo HJ, Yoon HH, Kim MY, Chung S, Kim YS, Chang YS, Park CW and Lee JH: Aggravation of diabetic nephropathy in BCL-2 interacting cell death suppressor (BIS)-haploinsufficient mice together with impaired induction of superoxide dismutase (SOD) activity. Diabetologia. 57:214–223. 2014. View Article : Google Scholar : PubMed/NCBI
46	Deshpande SD, Putta S, Wang M, Lai JY, Bitzer M, Nelson RG, Lanting LL, Kato M and Natarajan R: Transforming growth factor-β-induced cross talk between p53 and a microRNA in the pathogenesis of diabetic nephropathy. Diabetes. 62:3151–3162. 2013. View Article : Google Scholar : PubMed/NCBI
47	Kolati SR, Kasala ER, Bodduluru LN, Mahareddy JR, Uppulapu SK, Gogoi R, Barua CC and Lahkar M: BAY 11–7082 ameliorates diabetic nephropathy by attenuating hyperglycemia-mediated oxidative stress and renal inflammation via NF-κB pathway. Environ Toxicol Pharmacol. 39:690–699. 2015. View Article : Google Scholar : PubMed/NCBI
48	Gao F, Yao M, Shi Y, Hao J, Ren Y, Liu Q, Wang X and Duan H: Notch pathway is involved in high glucose-induced apoptosis in podocytes via Bcl-2 and p53 pathways. J Cell Biochem. 114:1029–1038. 2013. View Article : Google Scholar : PubMed/NCBI