Effect of rapamycin on autophagy of mesangial cells under high glucose condition
Objective To investigate the effect of rapamycin on autophagy of mesangial cells in high glucose state.Methods Human mesangial cells(HMC)were cultured in a constant tempera-ture incubater and divided into HMC group,low glycemic group,high glycemic group,mannitol iso-tonic group and rapamycin group in constant temperature incubator.The autophagy flow of cells in each group was marked by GFP-RFP-LC3.Western blot was used to detect the expression of autophagy re-lated proteins LC3-Ⅱ/LC3-Ⅰ and P62 in each group.The contents of cell type Ⅳ collagen fiber(ColⅣ),hyaluronic acid(HA)and laminin(LA)were determined by ELISA.Results Compared with HMC group,low glycemic group and mannitol isotonic group,LC3-Ⅱ expression was significantly de-creased,LC3-Ⅰ expression was significantly increased and P62 expression was significantly increased in high glycemic group.The expression of LC3-Ⅱ in rapamycin group increased significantly,while the expression of LC3-Ⅰ and P62 decreased significantly in high glycemic group,with statistical sig-nificance(all P＜0.05).The contents of Col Ⅳ,HA and LA in high glycemic group were significant-ly higher than those in HMC group(all P＜0.001).The contents of Col Ⅳ,HA and LA in rapamycin group were significantly decreased in high glycemic group,with statistical significance(all P＜0.05).Conclusions High glucose status can inhibit the autophagy activity of human mesangial cells,in-crease the aggregation of extracellular matrix,and promote the progression of diabetic nephropathy,while rapamycin can induce the occurrence of autophagy,reduce the proliferation of mesangial matrix of glomerular mesangial cells,and then inhibit the progression of diabetic nephropathy.
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