【Abstract】 Objective To construct eukaryotic expression vectors carrying the small hairpin RNA (shRNA) for NG2 mRNA and to study the effect on proliferation of rat mesangial cells (RMC) induced by high glucose stimulation through transfection and expression of shRNA targeting NG2 gene. Methods RMC was cultured in vitro and treated with high glucose (30 mmol/L) for 24 h. The change of NG2 expression was observed. Oligonucleotide hairpin sequences targeting NG2 gene were designed by the internet tool and was then inserted into the plasmid Pgenesil-1 containing enhanced green fluorescence protein (EGFP) to form plasmid Pgenesil-siNG21 and Pgenesil-siNG22. Plasmid expressing irrelevant shRNA was used as negative control named Pgenesil-siNC. RMC was transfected with above three kinds of plasmids respectively. Then the cells were stimulated with high glucose, and the changes of cell proliferation were observed by MTT and flow cytometry (FCM) methods. Results The expression of NG2 mRNA in RMC was significantly increased (P<0.01) after 24 h treatment with high glucose. The expression of EGFP gene was (50±10)%. The expression of NG2 in RMC was significantly inhibited by Pgenesil-siNG21 and Pgenesil-siNG22. Compared with the control group, cell proliferation levels were markedly decreased in siNG2 following high glucose stimulation (P<0.01). Conclusion shRNA targeting NG2 gene can inhibit the cell proliferation of RMC induced by high glucose stimulation.