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Nephrology (Carlton, Vic.)

Effects of stromal interaction molecule 1 or Orai1 overexpression on the associated proteins and permeability of podocytes.


PMID 26715123

Abstract

The present study was conducted to determine whether two important signalling molecules of store-operated channel (SOC), stromal interaction molecule 1 (STIM1) and Orai1, were involved in glomerular podocyte injury. We explored the effects of STIM1/Orai1 overexpression on podocyte associated proteins and cell permeability. The expression of STIM1 and Orai1 were examined in the renal cortex of adriamycin-induced nephropathy mice by real-time RT-PCR. The recombinant plasmid of STIM1/Orai1, identified by restriction enzyme digestion and PCR, was transfected into MPC5 cells via lipofectamine 2000. The transfecting efficiency was observed by a fluorescence microscope. RT-PCR and Western blotting were used to evaluate the expression levels of STIM1, Orai 1 and some podocyte-associated molecules in the transfected MPC5 cells. In addition, we examined the diffusion of FITC-dextran across the podocyte monolayer to investigate whether STIM1/Orai1 overexpression could affect cell permeability. We found that the mRNA levels of STIM1 and Orai1 were increased in adriamycin-induced nephropathy mice. STIM1/Orai1 overexpression significantly decreased the expression of podocin and CD2-associated protein (CD2AP), whereas it increased the expression of α-actinin-4. The permeability was significantly increased in the STIM1/Orai1 overexpression group. Our findings suggested that STIM1/Orai1 overexpression could affect the cell permeability and the expression of partial podocyte-associated proteins, which may ultimately result in podocyte injury.