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Effects of stromal interaction molecule 1 or Orai1 overexpression on the associated proteins and permeability of podocytes
Author(s) -
Miao Li,
Wei Dongyue,
Zhang Yuanyuan,
Liu Jiansheng,
Lu Siguang,
Zhang Aihua,
Huang Songming
Publication year - 2016
Publication title -
nephrology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.752
H-Index - 61
eISSN - 1440-1797
pISSN - 1320-5358
DOI - 10.1111/nep.12691
Subject(s) - podocyte , stim1 , orai1 , transfection , microbiology and biotechnology , lipofectamine , medicine , chemistry , biology , recombinant dna , kidney , biochemistry , endoplasmic reticulum , gene , proteinuria , vector (molecular biology)
Aim 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. Methods 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. Results 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. Conclusion 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.