ISSN 1006-298X      CN 32-1425/R

Chinese Journal of Nephrology, Dialysis & Transplantation

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Transcription factor KLF15 protects podocytes by suppressing NFATc1 signaling pathway

  

  • Online:2019-10-28 Published:2020-01-07

Abstract: Objective:To explore the molecular mechanism of Krüppel like Factor 15(KLF15)protecting podocytes by suppressing nuclear factor of activated Tcells cytoplasmic 1(NFATc1)signaling pathway.
Methodology:The expressions of KLF15 in podocytes of normal people and patients with glomerular disease were observed by immunofluorescence staining; Realtime quantitative PCR (RTPCR) and Western blot were used to detect expression of transcription factor KLF15 of the conditional immortalized mouse podocytes cultured in high glucose (HG) 30 mmol/L for 48h,lipopolysaccharide (LPS) 100 μg/ml for 48h and adriamycin (ADR) 025 μg/ml for 24h; After podocyte transfection of adenovirus transiently overexpressed KLF15,the effect of KLF15 overexpression,apoptosispromoting protein BAX and antiapoptotic protein BCL2 were detected by realtime  quantitative PCR (RTPCR) and Western blot; Apoptosis of podocytes was detected by flow cytometry after injury stimulation (ADR,LPS,HG) was administered to podocytes after overexpression KLF15; After  overexpression of KLF15 in podocytes,the expression of NFATc1 was detected by  RTPCR and Western blot; Binding of  KLF15 to NFATc1 promoter was detected by chromatinimmunoprecipitation (CHIP); The expression of NFATc1 downstream gene was detected by RTPCR.In vitro,podocytes were cultured and treated with dexamethasone,the expression of NFATc1 was observed by Western blot and RTPCR.
Results:In patients with glomerular disease,the expression of KLF15 in podocytes were decreased; The expression of KLF15  mRNA and protein were decreased in podocytes cultured in vitro; After overexpression of KLF15 in podocytes,the expression of proapoptotic protein BAX was decreased,and the expression of antiapoptotic protein BCL2 was increased.Under the condition of injury stimulation,the apoptotic rate in overexpression of KLF15 in podocytes was significantly lower than that in the control group; The expression of NFATc1 at mRNA and protein levels was decreased after overexpression of KLF15 in podocytes; Downstream target gene of NFATc1 was reduced.In normal podocytes,binding of KLF15  to the NFATc1 promoter region  was weakened by LPS stimulation,after overexpression of  KLF15 in podocytes,downstream genes of NFATc1 (Fzd9,Rcan1,Plaur)  mRNA were reduced.
Conclusion:
Transcription factor KLF15 protects podocytes by suppressing NFATc1,and dexamethasone may protect podocytes by inhibition of NFATc1 through increasing KLF15.

Key words: podocyte,Krü, ppel like factor 15, NFATc1 protein