Effects of recombinant lentivirus encoding human apM1 gene on platelet-derived growth factor inducedmesangial cell proliferation

Abstract

Objective To evaluate the effects of recombinant lentivirus encoding human apM1 gene ( LentiapM1-EGFP) on platelet-derived growth factor (PDGF) induced human mesangial cell (HMC) proliferation and intracellular AMP-activated protein kinase(AMPK) signaling pathway.Methods Protein expression of apM1 in cell culture supernatant of HMC transfected with Lenti-apM1-EGFP was detected by ELISA.The effect of human adiponectin on cell proliferation and cell cycle was assessed by [ 3 H ] thymidine incorporation assay and Flow cytometry respectively.The phosphorylation of AMPK was detected by Western blotting.Results Lenti-apM1-EGFP had no significant toxicity on HMC.The 50 multiplicity of infection (MOI) of the Lenti-apM1-EGFP efficiently infected HMC,and made it stable expression of adiponectin protein ( 149.6 ± 12.8 ) μg/L.PDGF-induced HMCs proliferation was significantly inhibited by adiponectin.When co-treatment with compound C,an AMPK inhibitor,the inhibitory effort was reversed.The phosphorylation level of AMPK was increased in HMC transfected Lenti-apM1-EGFP compared to that of control.Conclusions Adiponectin antagonizes stimulatory effect of platelet-derived growth factor on mesangial cell proliferation by AMPK signaling. Key words: Mesangial cell; Lentivirus; Adiponectin; AMP-activated protein kinase