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
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