Figure 2

Human adipose-derived mesenchymal stem cells (hAd-MSCs)-conditioned medium (CM) reduces apoptosis and injury of mouse podocyte clone (MPC5) cells induced by high glucose (HG). A) Representative photographs of annexinV/PI double-staining in different groups (n = 3 per group) and flow cytometry to test the podocytic apoptosis rate after culture in Wi38-CM or MSC-CM. MSC-CM reduced podocytic apoptosis in a dose-dependent manner, but Wi38-CM did not. B) Western blot was used to detect the expression of cleaved caspase-3 at three time points (24, 48 and 72 hours). MSC-CM reduced the expression of cleaved caspase-3, but Wi38-CM did not. All of the experiments were repeated three times (n = 3). *P <0.05 HG group versus normal glucose (NG) group or MSC-CM group; #P <0.05, 48-hour HG group or 72-hour HG group versus 24-hour HG group. C) The expression and location of the podocytic cytoskeletal protein synaptopodin (red) were measured by confocal microscopy. MSC-CM prevented the downregulation and rearrangement of synaptopodin, but Wi38-CM did not. Nuclei were stained with DAPI (blue). Magnification = 600×, 1800×. D) Using flow cytometry with TUNEL staining to measure the apoptosis rate of podocytes under treatment with NG, HG and MSCs-CM at three time points (24, 48 and 72 hours) (n = 3 each group). Cells analyzed under marker ‘A’ are apoptotic (TUNEL positive). The results showed that the podocytic apoptosis rate was significantly lower at all time points in the MSCs-CM group than in the HG group. DAPI, 4',6-diamidino-2-phenylindole; PI, propidium iodide.