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Cyclic AMP prevents decrease of phosphorylated ezrin/radixin/moesin and chloride intracellular channel 5 expressions in injured podocytes

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Abstract

Background

Our previous in vitro studies suggested that cyclin AMP (cAMP) signaling protects against podocyte injury. However, the molecular mechanisms remain unknown. The aim of the present study was to explore the role of forskolin, an agonist for adenylate cyclase, on ezrin/radixin/moesin (ERM) phosphorylation and chloride intracellular channel 5 (CLIC5) expressions in injured podocytes.

Methods

ADR nephrosis model were induced by adriamycin (ADR) injection in BalB/C mice. Parts of ADR nephrosis mice were pretreated with forskolin. Albuminuria was estimated by urine Coomassie blue stain. Nephrin, synaptopodin, CLIC5, phosphorylated ERM and podocalyxin were measured by confocal microscopy. CLIC5 and phosphorylated ERM also were studied using western blotting. RhoA and Rac1 were estimated by G-Lisa kit.

Results

We found that forskolin partially alleviated albuminuria and width of foot processes. Nephrin, synaptopodin, phosphorylated-ERM (p-ERM) and CLIC5 expression were decreased in ADR mice, which were improved by forskolin pretreatment. In vitro studies, pretreatment of podocytes with pCPT-cAMP(PKA-selective cAMP analogue)prevented puromycin aminonucleoside (PAN)-induced CLIC5 downregulation. 8-pCPT-2′-O-Me-cAMP (2Me-cAMP, an Epac-selective cAMP analogue) blocked PAN-induced p-ERM downregulation. PAN inhibited RhoA activation in podocytes, which could be prevented by pCPT-cAMP pretreatment. Y-27632, a Rho inhibitor, decreased CLIC5 expression in podocytes.

Conclusion

Activation cAMP signaling might attenuate albuminuria in ADR-induced nephrosis mice. Different downstream signaling pathway might mediate cAMP protection on CLIC5 and p-ERM expression, respectively.

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Acknowledgments

We are grateful to Professor Cijiang He (USA) for the podocyte clones. LG and ZN are supported by National Basic Research Program of China 973 Program 2012CB517602. LG is supported by National Nature Science Foundation Grant of China 30971363 and 81270781, as well as Nature Science Foundation Grant of Shanghai 08ZR1413200.

Conflict of interest

The authors have declared that no conflict of interest exists.

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    Authors and Affiliations

    1. Renal Division and Molecular Cell Lab for Kidney Disease, Renji Hospital, School of Medicine, Shanghai Jiaotong University, 160#, Pu Jian Road, Shanghai, 200001, China

      Hua Tao, Xiaoying Li, Kai Wei, Kewei Xie, Zhaohui Ni & Leyi Gu

    2. Renal Division, Kashgar Prefecture Second People’s Hospital of Xinjiang Uygur Autonomous Region, 1#, Jian Kang Road, Kashgar, Xinjiang Uygur Autonomous Region, China

      Leyi Gu

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    1. Hua Tao
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    Correspondence to Leyi Gu.

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    Hua Tao and Xiaoying Li have contributed equally for this paper.

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    Tao, H., Li, X., Wei, K. et al. Cyclic AMP prevents decrease of phosphorylated ezrin/radixin/moesin and chloride intracellular channel 5 expressions in injured podocytes. Clin Exp Nephrol 19, 1000–1006 (2015). https://doi.org/10.1007/s10157-015-1102-6

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    • DOI: https://doi.org/10.1007/s10157-015-1102-6

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