Effects of p53 on aldosterone-induced mesangial cell apoptosis in vivo and in vitro

Shi,Huimin; Zhang,Aiqing; He,Yanfang; Yang,Min; Gan,Weihua

doi:10.3892/mmr.2016.5156

Effects of p53 on aldosterone-induced mesangial cell apoptosis in vivo and in vitro

Authors:
- Huimin Shi
- Aiqing Zhang
- Yanfang He
- Min Yang
- Weihua Gan
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Affiliations: Department of Pediatric Nephrology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, Jiangsu 210011, P.R. China, Department of Nephrology, The Third Affiliated Hospital of Soochow University, Changzhou, Jiangsu 200040, P.R. China
Published online on: April 21, 2016 https://doi.org/10.3892/mmr.2016.5156
Pages: 5102-5108
Copyright: © Shi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Aldosterone (ALD) is a well‑known hormone, which may initiate renal injury by inducing mesangial cell (MC) injury in chronic kidney disease (CKD); however, the molecular mechanism remains unknown. The aim of the present study was to investigate the effects of p53 on ALD‑induced MC apoptosis and elucidate the underlying molecular mechanism. For the in vivo studies, rats were randomly assigned to receive normal saline or ALD for 4 weeks. The ratio of MC apoptosis was analysed by terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) assay. In addition, the expression level and localisation of p53, a well-known cell apoptosis-associated key protein, were detected by immunofluorescence. For the in vitro studies, rat MCs were incubated in medium containing either buffer (control) or ALD (10‑6 M) for 24 h. The cell apoptosis ratio was assessed by flow cytometry, and the expression level of p53 was assessed by reverse transcription quantitative polymerase chain reaction and western blotting. In order to confirm the role of p53 in ALD‑regulated cell apoptosis, a rescue experiment was performed using targeted small interfering (si)RNA to downregulate the expression of p53. The ALD‑treated rats exhibited greater numbers of TUNEL‑positive MCs and higher expression levels of p53 when compared with the control group. Furthermore, the ratio of MC apoptosis and the p53 expression level were significantly increased following ALD exposure, compared with the control group. Additionally, in the rescue experiment, the effects of ALD on MC were blocked by downregulating the expression level of p53 in MCs. The present study hypothesized that ALD may directly contribute to the occurrence of MC apoptosis via p53, which may participate in ALD-induced renal injury.

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1	White SL, Cass A, Atkins RC and Chadban SJ: Chronic kidney disease in the general population. Adv Chronic Kidney Dis. 12:5–13. 2005. View Article : Google Scholar : PubMed/NCBI
2	Lozano R, Naghavi M, Foreman K, Lim S, Shibuya K, Aboyans V, Abraham J, Adair T, Aggarwal R, Ahn SY, et al: Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: A systematic analysis for the global burden of disease study 2010. Lancet. 380:2095–2128. 2012. View Article : Google Scholar : PubMed/NCBI
3	Bolignano D, Palmer SC, Navaneethan SD and Strippoli GF: Aldosterone antagonists for preventing the progression of chronic kidney disease. Cochrane Database Syst Rev. 4:CD0070042014.PubMed/NCBI
4	Greene EL, Kren S and Hostetter TH: Role of aldosterone in the remnant kidney model in the rat. J Clin Invest. 98:1063–1068. 1996. View Article : Google Scholar : PubMed/NCBI
5	Lai L, Chen J, Hao CM, Lin S and Gu Y: Aldosterone promotes fibronectin production through a Smad2-dependent TGF-beta1 pathway in mesangial cells. Biochem Biophys Res Commun. 348:70–75. 2006. View Article : Google Scholar : PubMed/NCBI
6	Liang W, Chen C, Shi J, Ren Z, Hu F, van Goor H, Singhal PC and Ding G: Disparate effects of eplerenone, amlodipine and telmisartan on podocyte injury in aldosterone-infused rats. Nephrol Dial Transplant. 26:789–799. 2011. View Article : Google Scholar :
7	Del Vecchio L, Procaccio M, Vigano S and Cusi D: Mechanisms of disease: The role of aldosterone in kidney damage and clinical benefits of its blockade. Nat Clin Pract Nephrol. 3:42–49. 2007. View Article : Google Scholar
8	Nishiyama A, Yao L, Nagai Y, Miyata K, Yoshizumi M, Kagami S, Kondo S, Kiyomoto H, Shokoji T, Kimura S, et al: Possible contributions of reactive oxygen species and mitogen-activated protein kinase to renal injury in aldosterone/salt-induced hypertensive rats. Hypertension. 43:841–848. 2004. View Article : Google Scholar : PubMed/NCBI
9	Fan YY, Baba R, Nagai Y, Miyatake A, Hosomi N, Kimura S, Sun GP, Kohno M, Fujita M, Abe Y and Nishiyama A: Augmentation of intrarenal angiotensin II levels in uninephrectomized aldosterone/salt-treated hypertensive rats; renoprotective effects of an ultrahigh dose of olmesartan. Hypertens Res. 29:169–178. 2006. View Article : Google Scholar : PubMed/NCBI
10	Kiyomoto H, Rafiq K, Mostofa M and Nishiyama A: Possible underlying mechanisms responsible for aldosterone and mineralocorticoid receptor-dependent renal injury. J Pharmacol Sci. 108:399–405. 2008. View Article : Google Scholar : PubMed/NCBI
11	Shibata S, Nagase M, Yoshida S, Kawachi H and Fujita T: Podocyte as the target for aldosterone: Roles of oxidative stress and Sgk1. Hypertension. 49:355–364. 2007. View Article : Google Scholar : PubMed/NCBI
12	Nagase M and Fujita T: Endocrinological aspects of proteinuria and podocytopathy in diabetes: Role of the aldosterone/mineralocorticoid receptor system. Curr Diabetes Rev. 7:8–16. 2011. View Article : Google Scholar
13	Epstein M: Aldosterone blockade: An emerging strategy for abrogating progressive renal disease. Am J Med. 119:912–919. 2006. View Article : Google Scholar : PubMed/NCBI
14	Zhu D, Yu H, He H, Ding J, Tang J, Cao D and Hao L: Spironolactone inhibits apoptosis in rat mesangial cells under hyperglycaemic conditions via the Wnt signalling pathway. Mol Cell Biochem. 380:185–193. 2013. View Article : Google Scholar : PubMed/NCBI
15	Nagase M and Fujita T: Aldosterone and glomerular podocyte injury. Clin Exp Nephrol. 12:233–242. 2008. View Article : Google Scholar : PubMed/NCBI
16	Merkel CE, Karner CM and Carroll TJ: Molecular regulation of kidney development: Is the answer blowing in the Wnt? Pediatr Nephrol. 22:1825–1838. 2007. View Article : Google Scholar : PubMed/NCBI
17	Chen C, Liang W, Jia J, van Goor H, Singhal PC and Ding G: Aldosterone induces apoptosis in rat podocytes: Role of PI3-K/Akt and p38MAPK signaling pathways. Nephron Exp Nephrol. 113:e26–e34. 2009. View Article : Google Scholar : PubMed/NCBI
18	Abboud HE: Mesangial cell biology. Exp Cell Res. 318:979–985. 2012. View Article : Google Scholar : PubMed/NCBI
19	Brunskill EW and Potter SS: Changes in the gene expression programs of renal mesangial cells during diabetic nephropathy. BMC Nephrol. 13:702012. View Article : Google Scholar : PubMed/NCBI
20	Dalla Vestra M, Saller A, Mauer M and Fioretto P: Role of mesangial expansion in the pathogenesis of diabetic nephropathy. J Nephrol. 14(Suppl 4): S51–S57. 2001.
21	Kitamura H, Shimizu A, Masuda Y, Ishizaki M, Sugisaki Y and Yamanaka N: Apoptosis in glomerular endothelial cells during the development of glomerulosclerosis in the remnant-kidney model. Exp Nephrol. 6:328–336. 1998. View Article : Google Scholar : PubMed/NCBI
22	Shimizu A, Masuda Y, Kitamura H, Ishizaki M, Sugisaki Y and Yamanaka N: Apoptosis in progressive crescentic glomerulonephritis. Lab Invest. 74:941–951. 1996.PubMed/NCBI
23	Mishra R, Emancipator SN, Kern T and Simonson MS: High glucose evokes an intrinsic proapoptotic signaling pathway in mesangial cells. Kidney Int. 67:82–93. 2005. View Article : Google Scholar
24	Sugiyama H, Kashihara N, Makino H, Yamasaki Y and Ota A: Apoptosis in glomerular sclerosis. Kidney Int. 49:103–111. 1996. View Article : Google Scholar : PubMed/NCBI
25	Ford HL, Sclafani RA and DeGregori J: Cell cycle and growth control: Biomolecular regulation and cancer. Stein GSaP AB: Cell cycle regulatory cascades Hoboken, New Jersey: Wiley-Liss; pp. 95–128. 2004
26	Livak and Schmittgen: Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCt method. Methods. 25:402–408. 2001. View Article : Google Scholar
27	Chrysostomou A and Becker G: Spironolactone in addition to ACE inhibition to reduce proteinuria in patients with chronic renal disease. N Engl J Med. 345:925–926. 2001. View Article : Google Scholar : PubMed/NCBI
28	Rocha R, Chander PN, Zuckerman A and Stier CT Jr: Role of aldosterone in renal vascular injury in stroke-prone hypertensive rats. Hypertension. 33:232–237. 1999. View Article : Google Scholar : PubMed/NCBI
29	Blasi ER, Rocha R, Rudolph AE, Blomme EA, Polly ML and McMahon EG: Aldosterone/salt induces renal inflammation and fibrosis in hypertensive rats. Kidney Int. 63:1791–1800. 2003. View Article : Google Scholar : PubMed/NCBI
30	Miric G, Dallemagne C, Endre Z, Margolin S, Taylor SM and Brown L: Reversal of cardiac and renal fibrosis by pirfenidone and spironolactone in streptozotocin-diabetic rats. Br J Pharmacol. 133:687–694. 2001. View Article : Google Scholar : PubMed/NCBI
31	Aldigier JC, Kanjanbuch T, Ma LJ, Brown NJ and Fogo AB: Regression of existing glomerulosclerosis by inhibition of aldosterone. J Am Soc Nephrol. 16:3306–3314. 2005. View Article : Google Scholar : PubMed/NCBI
32	Han SY, Chang EJ, Choi HJ, Kwak CS, Park SB, Kim HC and Mun KC: Apoptosis by cyclosporine in mesangial cells. Transplant Proc. 38:2244–2246. 2006. View Article : Google Scholar : PubMed/NCBI
33	Oltvani ZN and Korsmeyer SJ: Checkpoints of dueling dimers foli death wishes. Cell. 79:189–192. 1994. View Article : Google Scholar
34	Vogelstein B and Kinzler KW: p53 function and dysfunction. Cell. 70:523–526. 1992. View Article : Google Scholar : PubMed/NCBI
35	Miyashita T and Reed JC: Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell. 80:293–299. 1995. View Article : Google Scholar : PubMed/NCBI
36	Pierzchalski P, Reiss K, Cheng W, Cirielli C, Kajstura J, Nitahara JA, Rizk M, Capogrossi MC and Anversa P: p53 induces myocyte apoptosis via the activation of the renin-angiotensin system. Exp Cell Res. 234:57–65. 1997. View Article : Google Scholar : PubMed/NCBI
37	Leri A, Fiordaliso F, Setoguchi M, Limana F, Bishopric NH, Kajstura J, Webster K and Anversa P: Inhibition of p53 function prevents renin-angiotensin system activation and stretch-mediated myocyte apoptosis. Am J Pathol. 157:843–857. 2000. View Article : Google Scholar : PubMed/NCBI
38	el-Deiry WS: Regulation of p53 downstream genes. Semin Cancer Biol. 8:345–357. 1998. View Article : Google Scholar
39	Chang NS: A potential role of p53 and WOX1 in mitochondrial apoptosis (review). Int J Mol Med. 9:19–24. 2002.