Abstract: Objective: To investigate the preparation process of Lycopus lucidus polysaccharides （LLP） with different ethanol
precipitation fractions and evaluate their effects on high glucose-induced rat glomerular mesangial cells （HBZY-1）. Methods: The
crude LLP was obtained by deproteinization and macroporous resin column chromatography, and the components were obtained by
fractional ethanol precipitation. The activity of high glucose-induced HBZY-1 cells was tested in vitro. Results: Three different
ethanol precipitation fractions （LLP-30%, LLP-60%, and LLP-90%） showed no toxicity to normal HBZY-1 cells at concentrations
ranging from 0.25 to 4.00 mg/mL. LLP-60% and LLP-90% exhibited significant inhibitory effects on the proliferation of high glucoseinduced
HBZY-1 cells （P<0.01） and promoted the content of superoxide dismutase while reducing the contents of malondialdehyde
and reactive oxygen species. Conclusion: The extraction, separation and preparation process of LLP are feasible, and the active
component LLP-90% not only has the highest content, but also exhibits the most significant effects on inhibiting the proliferation of
high glucose-induced HBZY-1 cells and alleviating cellular oxidative stress, which provided the basis for the development of L.
lucidus as a functional food of diabetic nephropathy.

Key words: Lycopus lucidus polysaccharides, fractional ethanol precipitation, glomerular mesangial cells, proliferation, oxidative stress