Chinese Journal of Nephrology, Dialysis & Transplantation
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Abstract: Objective:To explore the molecular mechanism of Krüppel like Factor 15(KLF15)protecting podocytes by suppressing nuclear factor of activated Tcells 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; Realtime quantitative PCR (RTPCR) 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) 025 μg/ml for 24h; After podocyte transfection of adenovirus transiently overexpressed KLF15,the effect of KLF15 overexpression,apoptosispromoting protein BAX and antiapoptotic protein BCL2 were detected by realtime quantitative PCR (RTPCR) 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 RTPCR and Western blot; Binding of KLF15 to NFATc1 promoter was detected by chromatinimmunoprecipitation (CHIP); The expression of NFATc1 downstream gene was detected by RTPCR.In vitro,podocytes were cultured and treated with dexamethasone,the expression of NFATc1 was observed by Western blot and RTPCR. 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 proapoptotic protein BAX was decreased,and the expression of antiapoptotic 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
DOU Caoshuai,ZHANG Hong,KE Guibao, et al. Transcription factor KLF15 protects podocytes by suppressing NFATc1 signaling pathway[J]. Chinese Journal of Nephrology, Dialysis & Transplantation, DOI: 10.3969/j.issn.1006-298X.2019.05.006.
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URL: http://www.njcndt.com/EN/10.3969/j.issn.1006-298X.2019.05.006
http://www.njcndt.com/EN/Y2019/V28/I5/427