Journal of Shandong University (Health Sciences) ›› 2022, Vol. 60 ›› Issue (5): 16-21.doi: 10.6040/j.issn.1671-7554.0.2021.1504

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Effects of orphan nuclear receptor NR4A1 on H2O2 induced injury of mouse renal podocytes

LIU Min1, ZHANG Yuchao2, MA Xiaoli2, LIU Xinyu2, SUN Lu2, ZUO Dan2, LIU Yuantao3   

  1. 1. Qingdao Clinical Medical School of Nanjing Medical University, Qingdao Municipal Hospital, Qingdao 266011, Shandong, China;
    2. Department of Endocrinology, Qingdao Municipal Hospital, Qingdao 266011, Shandong, China;
    3. Department of Endocrinology, Qilu Hospital of Shandong University, Qingdao 266011, Shandong, China
  • Published:2022-06-01

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Abstract: Objective To explore the expression of orphan nuclear receptor(NR4A1)in mouse renal podocytes(MPC-5)under H2O2 induced oxidative stress and to investigate the effects of NR4A1 on the viability and apoptosis of MPC-5. Methods MPC-5 were divided into blank control group(H2O2 concentration was 0 μmol/L or treatment time was 0 h), and treatment groups(related to different H2O2 concentrations or treatment time). After MPC-5 were treated with H2O2 to establish the oxidative stress model, the mRNA and protein expressions of NR4A1 and Caspase 3(17 kDa)were detected with Q-PCR and Western blotting, respectively. After MPC-5 were infected with lentivirus with cDNA encoding NR4A1, the stable cell lines with NR4A1 over-expression(Ad-NR4A1 group)and the empty vector(Ad-Ctrl group)were selected with puromycin. After H2O2 treatment, the protein expression of Caspase 3(17 kDa), viability and apoptosis were detected with Western blotting, CCK8 assay and TUNEL staining, respectively. Results After H2O2 treatment, both protein and mRNA expressions of NR4A1 were significantly higher in the treatment groups than in the blank control group(P<0.05), and the protein expression of Caspase 3(17 kDa)also significantly increased(P<0.05). The protein expression of NR4A1 was much higher in Ad-NR4A1 group than in Ad-Ctrl group(P<0.001). After H2O2 treatment, the protein expression of caspase 3(17 kDa)was significantly higher in Ad-NR4A1 group than in Ad-Ctrl group, the apoptotic rate was significantly higher(P<0.05), and the cell viability was significantly lower(P<0.05). Conclusion H2O2 induced oxidative stress contributes to the decreased cell viability and increased cell apoptosis in MPC-5. The possible mechanism is related to the activation of Caspase 3 by NR4A1.

Key words: Mouse renal podocytes, Orphan nuclear receptor NR4A1, Oxidative stress, Apoptosis

CLC Number: 

  • R34
[1] Ghaderian SB, Hayati F, Shayanpour S, et al. Diabetes and end-stage renal disease; a review article on new concepts [J]. J Renal Inj Prev, 2015, 4(2): 28-33.
[2] Lim AKh. Diabetic nephropathy-complications and treatment [J]. Int J Nephrol Renovasc Dis, 2014, 7: 361-381. doi: 10.2147/IJNRD.S40172.
[3] Liu BC, Song X, Lu XY, et al. High glucose induces podocyte apoptosis by stimulating TRPC6 via elevation of reactive oxygen species [J]. Biochim Biophys Acta, 2013, 1833(6): 1434-1442.
[4] Bose M, Almas S, Prabhakar S. Wnt signaling and podocyte dysfunction in diabetic nephropathy [J]. J Investig Med, 2017, 65(8): 1093-1101.
[5] Zhang XK. Targeting Nur77 translocation[J]. Expert Opin Ther Targets, 2007, 11(1): 69-79.
[6] Pearen MA, Muscat GEO. Minireview: nuclear hormone receptor 4A signaling: implications for metabolic disease [J]. Mol Endocrinol, 2010, 24(10): 1891-1903.
[7] Ogawa D, Eguchi J, Wada J, et al. Nuclear hormone receptor expression in mouse kidney and renal cell lines [J]. PLoS One, 2014, 9(1): e85594.
[8] Sheng J, Li H, Dai Q, et al. NR4A1 promotes diabetic nephropathy by activating Mff-mediated mitochondrial fission and suppressing parkin-Mediated mitophagy [J]. Cell Physiol Biochem, 2018, 48(4): 1675-1693.
[9] Shankland SJ. The podocytes response to injury: role in proteinuria and glomerulosclerosis [J]. Kidney Int, 2006, 69(12): 2131-2147.
[10] Pagtalunan ME, Miller PL, Jumping-Eagle S, et al. Podocyte loss and progressive glomerular injury in type II diabetes [J]. J Clin Invest, 1997, 99(2): 342-348.
[11] Kriz W, Gretz N, Lemley KV. Progression of glomerular diseases: is the podocyte the culprit? [J]. Kidney Int, 1998, 54(3): 687-697.
[12] Lee SO, Jin UH, Kang JH, et al. The orphan nuclear receptor NR4A1(Nur77)regulates oxidative and endoplasmic Reticulum stress in pancreatic cancer cells [J]. Mol Cancer Res, 2014, 12(4): 527-538.
[13] Zong C, Qin D, Yu C, et al. The stress-response molecule NR4A1 resists ROS-induced pancreatic β-cells apoptosis via WT1 [J]. Cell Signal, 2017, 35: 129-139. doi: 10.1016/j.cellsig.2017.03.012.
[14] Herring JA, Elison WS, Tessem JS. Function of NR4A orphan nuclear receptors in proliferation, apoptosis and fuel utilization across tissues [J]. Cells, 2019, 8(11): 1373.
[15] 吴凌娟, 徐晓丽, 林峻, 等. 核受体参与调控细胞自噬的研究进展[J]. 生物医学工程学杂志, 2018, 35(5): 822-828.
[16] Pearen MA, Muscat GE. Minireview: nuclear hormone receptor 4A signaling: implications for metabolic disease [J]. Mol Endocrinol, 2010, 24(10): 1891-1903.
[17] Safe S, Karki K. The paradoxical roles of orphan nuclear receptor 4A(NR4A)in cancer [J]. Mol Cancer Res, 2021, 19(2): 180-191.
[18] Pu ZQ, Yu TF, Liu D, et al. NR4A1 enhances MKP7 expression to diminish JNK activation induced by ROS or ER-stress in pancreatic β cells for surviving [J]. Cell Death Discov, 2021, 7(1): 133.
[19] Munnur D, Somers J, Skalka G, et al. NR4A nuclear receptors target poly-ADP-ribosylated DNA-PKcs protein to promote DNA repair [J]. Cell Rep, 2019, 26(8): 2028-2036.
[20] Chen HZ, Wen Q, Wang WJ, et al. The orphan nuclear receptor TR3/Nur77 regulates ER stress and induces apoptosis via interaction with TRAPγ [J]. Int J Biochem Cell Biol, 2013, 45(8): 1600-1609.
[21] Holmes WF, Soprano DR, Soprano KJ. Early events in the induction of apoptosis in ovarian carcinoma cells by CD437: activation of the p38 MAP kinase signal pathway [J]. Oncogene, 2003, 22(41): 6377-6386.
[22] Nuñez G, Benedict MA, Hu Y, et al. Caspases: the proteases of the apoptotic pathway [J]. Oncogene, 1998, 17(25): 3237-3245.
[23] Mazumder S, Plesca D, Almasan A. Caspase-3 activation is a critical determinant of genotoxic stress-induced apoptosis [J]. Methods Mol Biol, 2008, 414: 13-21. doi: 10.1007/978-1-59745-339-4_2.
[24] Lin B, Kolluri SK, Lin F, et al. Conversion of Bcl-2 from protector to killer by interaction with nuclear orphan receptor Nur77/TR3 [J]. Cell, 2004, 116(4): 527-540.
[25] Porter AG, Jänicke RU. Emerging roles of caspase-3 in apoptosis [J]. Cell Death Differ, 1999, 6(2): 99-104.
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