Background: Very-low-density lipoprotein (VLDL) in vitro can induce foam cell formation in human mesangial cells. Lipoprotein lipase (LPL) expressed in the arterial wall plays a key role in atherogenesis by actions of enzymolysis and ‘molecular bridge’, and, thereby, leads to the formation of lipid-loaded foam cells. It is known that LPL is expressed by glomerular mesangial cells. This study was designed to investigate if LPL plays a role in VLDL-induced lipid accumulation in human mesangial cells and its underlying mechanism. Methods: Human wild-type LPL (hLPLwt), catalytically inactive LPL (hLPL194) or control alkaline phosphatase (hAP) were expressed in human mesangial cell line (HMCLs) via adenoviral vectors. Orlistat (tetrahydrolipstatin), a specific inhibitor of the lipoidolytic activity of endogenous LPL, and heparinase, which degrades cell-surface heparan sulfate proteoglycans, were also used to estimate the role of either the enzymolysis or ‘molecular bridge’ actions of LPL in the uptake of VLDL. Anti-low-density lipoprotein receptor (LDLr) antibody and anti-LDL receptor-related protein antibody were used to evaluate the effect of lipoprotein receptors on VLDL-induced lipid accumulation in HMCLs. Cellular lipid deposition was visualized by Oil Red O staining and analyzed quantitatively by standard enzymatic procedures. LPL protein expression and activity were measured by Western blot and a chemical analysis, respectively. Results: VLDL induced triglyceride accumulation in HMCLs in a time- and dose-dependent manner. Compared with Ad-hAP transfected HMCLs, cellular triglyceride content increased 4.55-fold (p < 0.05) in Ad-hLPLwt-transfected HMCLs and 1.52-fold (p < 0.05) in Ad-hLPL194-transfected HMCLs. Triglyceride accumulation in response to VLDL was mostly blocked by orlistat. Pretreatment of the cells with heparinase slightly reduced cellular triglyceride accumulation in the present of high concentrations of VLDL. The blockade of some lipoprotein receptors, such as LDLr, did not significantly reduce cellular triglyceride accumulation. LPL expression was upregulated by VLDL. Conclusions: VLDL-induced triglyceride accumulation in human mesangial cells is mainly mediated by LPL, and the enzymolysis action of LPL could be a major factor in this process. These results suggest that LPL may be an important factor participating in the initiation and progression of VLDL-mediated lipid renal injury.