Objective To explore the effect of silent information regulator 1（SIRT1） on acetylation of nuclear factor-κB (NF-κB) p65 subunit and the expression of monocyte chemoattractant protein 1 (MCP-1) in rat mesangial cells (RMC) induced by high glucose. Methods Construct the lentiviral shRNA plasmid pTRC-shSIRT1 to interference SIRT1 gene and identification . The RMCs was classified into high glucose group (high glucose culture medium used), resveratrol high glucose group (with low glucose culture medium containing 1μmol / L SIRT1 activator resveratrol for 24 h and then switching to high glucose culture medium), SIRT1 RNAi group (4h after viral pTRC-shSIRT1 infection, and then switch to low-glucose culture medium), SIRT1 RNAi high glucose group (4h after viral pTRC-shSIRT1 infection, and then switch to high glucose culture medium) , While negative control group and the control group (hypertonic mannitol). The mRNA expression of SIRT1 and MCP-1 gene was analyzed by real-time quantitative PCR. The protein expression of SIRT1 and the acetylation of NF-κB p65 subunit were observed by Western Blot. The protein expression of MCP-1 was detected by ELISA. Results DNA sequencing demonstrated that the plasmid pTRC-shSIRT1 was successfully constructed. Viral produced by pTRC-shSIRT1 infected RMCs and knocked down SIRT1 expression both at mRNA and protein levels. The expression of SIRT1 was decreased and acetylation of NF-κB p65 subunit was significantly increased by high glucose. Resveratrol, as an activator of SIRT1 can decreases high glucose-induced acetylation of NF-κB p65, which results in the reduction of MCP-1 secretion. Conversely, Gene silencing of SIRT1 shows increased the acetylation of cellular p65 protein and secretion of MCP-1 when challenged with high glucose. Conclusion The plasmid pTRC-shSIRT1 was successfully constructed and can effectively down regulate SIRT1 mRNA and protein expression. SIRT1 activation can significantly inhibited high glucose mediated upregulation of MCP-1, This is probably attributable to the increasing SIRT1-mediated NF-κB p65 deacetylation.