Original article (Basic research)

Prevention of apoptosis of podocytes by protein kinase A via cAMP response element binding protein signaling pathway

  • XIANG Peng ,
  • XIE Ke-wei ,
  • JIANG Jin-xing ,
  • et al
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  • Department of Nephrology, Molecular Cell Laboratory for Kidney Disease, Renji Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200127, China

Online published: 2016-02-26

Supported by

National Natural Science Foundation of China, 30971363, 81270781。

Abstract

Objective To explore the effects of cAMP response element binding protein (CREB) signaling pathway on the protection of podocytes by the protein kinase A (PKA). Methods The 14-25 generations of mouse podocytes 5P12 differentiated under constant condition were selected and divided into the control group, adriamycin (ADR) treatment group (ADR group), and PKA specific agonist (pCPT-cAMP) pre-culture+ADR stimulation group (pCPT-cAMP+ADR group). Cell toxicity was detected by CCK-8 method. The expression of cleaved caspase-3 in podocytes was detected by Western blotting. The expression of CREB in podocytes was inhibited by siRNA and the localization and expression of phosphorylated CREB (p-CREB) were detected by immunofluorescence confocal microscopy and Western blotting. Results ADR induced up-regulation of the expression of cleaved caspase-3 in podocytes and pre-culture by pCPT-cAMP could significantly decrease the elevated expression of cleaved caspase-3 induced by ADR. ADR decreased the number of podocytes to (40.45±6.78)% of its original number and pre-culture by pCPT-cAMP increased the number of podocytes to (69.8±5.48)% of its original number. Results of Western blotting showed that pre-culture by pCPT-cAMP for 5 and 15 min could increase the expressions of p-CREB in total protein of podocytes by (105.64±38.21)% and (98.61±41.03)% and those in nuclei by (114.52±11.28)% and (118.84±14.44)%. Immunofluorescence confocal microscopy showed that pCPT-cAMP mainly increased the expression of p-CREB in nuclei. siRNA could decreased the protein expression of CREB in podocytes to (25.1±2.44)% of its original expression and pre-culture by pCPT-cAMP could not prevent the ADR-induced up-regulation of the expression of cleaved caspase-3. Conclusion Transcription factor CREB mediates the protective effect of PKA signaling pathway on podocytes.

Cite this article

XIANG Peng , XIE Ke-wei , JIANG Jin-xing , et al . Prevention of apoptosis of podocytes by protein kinase A via cAMP response element binding protein signaling pathway[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2016 , 36(1) : 9 . DOI: 10.3969/j.issn.1674-8115.2016.01.002

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