The Effect Of High Glucose On The Wnt-β-catenin Signaling Pathway In Human Mesangial Cells And The Interventive Effect Of Rhein

Objective:To investigate the effect of high glucose on the Wnt/β-catenin signaling pathway in human mesangial cells, and to explore the intervention effect and the underlying mechanisms of rhein.Methods:Human mesangial cells were revived,cultured respectively in DMEM and10%FBS in a5%CO2,37℃incubator for several days and harvested the logarithmic growth phase cells for further studies. Incubated the cells for24h to synchronize with fetal bovine serum-free medium when there were about80%adherented. Then divided the cells into the following groups. NG (5.6mmol/L) group, HG (30mmol/L) group,HG (30mmol/L)+Rhein (Rhein110-4mol/L, Rhein210-5mol/L, Rhein310-6mol/L) group, Measured the proliferative activity of cells by the MTT colorimetry after cultured24h,48h and72h. Collecting the HMC after cultured48h and observing the expression of Wnt, β-catenin in mRNA by using RT-PCR; The expression of Wnt、β-catenin protein was detected by using immunohistochemistry. The expression of TGF-β1was examined by ELISA.Results:Compared with the NG group, the HG group and HG+Rhein group OD values increased significantly (P<0.05); compared with the HG group, HG+Rhein group showed an downward trend in24h, but not significantly (P>0.05) And in48h and72h, the downward trend is significant, expressed as time, concentration-dependent (P<0.05or P<0.01); Compared with the HG+Rhein3group, HG+Phein2group showed an downward trend significantly, expressed as time, concentration-dependent (P<0.05), the HG+Rhein1group discreased significantly in48h and72h (P<0.05); Compared with the HG+Rhein2group, HG+Rhein1group showed an downward trend significantly in72h (P<0.05). In the nomal state, the mesangial cells expressed a certain amount of Wnt and β-catenin, when they were stimulated by high glucose, the expression of Wnt-(3-catenin in mRNA and protein increased (P<0.05);Rhein significantly reduced the expression of them in high glucose-induced mesangial cells (P<0.05). The high glucose groups could promote cells to secrete TGF-β1, In HG+Rhein group, the secretion was decreased (P<0.05).Conclusion:In vitro, rhein could inhibit the proliferation of mesangial cells which were promoted by hyperglycemia, the mechanism of which may be related to down-regulating the expression of Wnt, β-catenin and the secretion of TGF-β1.