Trehalose decreased PAN-induced apoptosis in human podocytes.

<p>Podocyte were treated with PAN (30 µg/ml) or/and Trehalose (50 mM) for 48 h. (<b>A</b>) Trehalose induced autophagy in PAN-treated human podocytes. The expression of LC3-II slightly increased after 48 h PAN treatment, while it dramatically up-regulated in Tre and PAN+ Tre groups. Representative immunoblot images were shown along with the statistical results. **<i>p</i><0.01 versus CON, n = 6. (<b>B–C</b>) The findings of (A) were confirmed by LC3 immunostaining. Obvious elevated LC3-II bright green puncta (indicated by white arrows) were visualized in trehalose-treated groups (Tre and PAN+ Tre groups), the representative images and statistical results were shown. Nuclei were stained in blue. **<i>p</i><0.01, ***<i>p</i><0.001 versus CON, n = 6. (<b>D</b>) No significant changes were observed in podocyte necrosis. LDH in culture medium of 4 groups was measured, n = 4. (<b>E</b>) Elevated apoptosis in PAN-treated podocytes was decreased by trehalose. Apoptosis was measured by flow cytometry with YO-PRO-1/PI assay. Podocyte apoptosis was induced by PAN and decreased significantly by trehalose. *<i>p</i><0.05, **<i>p</i><0.01 versus CON, n = 8. (<b>F</b>) The findings of (E) were confirmed by the data of active caspase-3 measurement. The active caspase-3 positive podocytes were measured by flow cytometry. The changes pattern was similar to podocyte apoptosis measured by YO-PRO-1/PI assay. **<i>p</i><0.01 versus CON, n = 7.