〔Abstract〕 Objective To investigate the effect of berberine(BBR) on the proliferation and autophagy of mesangial cells in high glucose(HG) environment and the specific molecular mechanism. Methods Mesangium cells at exponential growth stage were divided into the following groups: normal group, high glucose group, high glucose + BBR treatment group(30, 60 and 90 μmol/L), high glucose + BBR(90 μmol/L) + AMPK inhibitor Compound C group(CC group); the number of mesangial cells was calculated by high content cell imager. The expressions of type Ⅳ collagen(Col-Ⅳ), fibronectin(FN) and microtubule-associated protein 1 light chain 3B(LC3B) in mesangial cells were detected by immunofluorescence assay. The protein expression levels of LC3B, Beclin-1, p62, Col-Ⅳ, FN and silencing regulatory factor 1(SIRT1)/adenylate activated protein kinase(AMPK) signaling pathway were detected by Western blot. Results Compared with the normal group, high content cell imaging showed abnormal proliferation of mesangial cells in the hyperglycemic group. The results of immunofluorescence and Western blot showed that the expression levels of Col-Ⅳ and FN deposited in mesangial extracellular matrix increased in the high glucose group. The results of Western blot showed that the protein expressions of SIRT1, p-AMPK, LC3B and Beclin-1 decreased, while the protein expressions of p-p65 and p62 increased. BBR inhibited the abnormal proliferation of mesangial cells induced by high glucose. BBR could reduce the expression levels of Col-Ⅳ and FN deposited in mesangial extracellular matrix. BBR could increase the expressions of SIRT1, p-AMPK, LC3B and Beclin-1 proteins in mesangial cells, while decrease the expressions of p-p65 and p62 proteins. CC group weakened the inhibition of mesangial cell proliferation and autophagy by high dose BBR. Conclusion Berberine can effectively inhibit the proliferation of mesangial cells induced by high glucose and increase the level of autophagy, which may be related to SIRT1/AMPK signaling pathway.