摘要
Objective:To study the potential effect of miR-21 and SIRT1 on CD40 expression induced by TNF-α and underlying mechanisms in renal inner medulla collecting duct(IMCD)cells.Method:After stimulation with the indicated times and doses of TNF-α,miR-21 mRNA levels were analyzed by RT-qPCR.Following transfection with miR-21 mimics or miR-21 inhibitor,the mRNA and protein expression of SIRT1 and CD40 were identified by RT-qPCR and Western blot,respectively.Furthermore,the underlying molecular mechanism was also investigated.Results:TNF-α increased miR-21,CD40 and Ac-p65 and reduced SIRT1 expression.miR-21 mimics increased SIRT1 and reduced Ac-p65 and CD40 expressions,the corresponding changes were observed by miR-21 inhibitor,which was impeded by pre-treating with SIRT1 siRNA.Overexpression of SIRT1 reduced CD40 and Ac-p65 expression,which was reversed by silencing SIRT1 and augmented by NF-κB siRNA.Conclusion:miR-21 inhibits TNF-α-induced CD40 expression in IMCD cells via the SIRT1-NF-κB signaling pathway.
Objective:To study the potential effect of miR-21 and SIRT1 on CD40 expression induced by TNF-α and underlying mechanisms in renal inner medulla collecting duct(IMCD)cells.Method:After stimulation with the indicated times and doses of TNF-α,miR-21 mRNA levels were analyzed by RT-qPCR.Following transfection with miR-21 mimics or miR-21 inhibitor,the mRNA and protein expression of SIRT1 and CD40 were identified by RT-qPCR and Western blot,respectively.Furthermore,the underlying molecular mechanism was also investigated.Results:TNF-α increased miR-21,CD40 and Ac-p65 and reduced SIRT1 expression.miR-21 mimics increased SIRT1 and reduced Ac-p65 and CD40 expressions,the corresponding changes were observed by miR-21 inhibitor,which was impeded by pre-treating with SIRT1 siRNA.Overexpression of SIRT1 reduced CD40 and Ac-p65 expression,which was reversed by silencing SIRT1 and augmented by NF-κB siRNA.Conclusion:miR-21 inhibits TNF-α-induced CD40 expression in IMCD cells via the SIRT1-NF-κB signaling pathway.
引文