Influence of Nrf2/HO-l/GPX4 signaling pathway on high glucose-induced podocyte ferroptosis and intervention of berberine / 中国药理学通报

ABSTRACT

Aim To explore the regulatory mechanism of berberine in the high glucose (HG) -induced podocyte ferroptosis. Methods Western blot and R T - qPCR were used to detect the changes of G P X 4, PTGS2, ACSL4, podocin, and desmin at different time (0 h, 6 h, 12 h, 24 h, 36 h) of HG stimulation, and the relative expression of Nrf2, HO-1, GPX4 and podocin under HG after treated with berberine. The proliferation of podocytes stimulated by HG at different time points was observed by EdU staining. The therapeutic effect of berberine on podocyte damage was screened by CCK-8, and the effect of berberine on the level of oxidative stress in HG-induced podocytes were measured by fluorescence inverted microscope, GSH and GSSG kits. In addition, transmission electron microscopy was employed to observe the ultrastructural characteristics of podocytes in each group. Results The expression of podocin and GPX4 was significantly reduced at 24 h, and the mRNA levels of ACSL4 and PTGS2 were significantly up-regulated. Berberine could notably increase the expression of Nrf2, HO-1, GPX4 and podocin, and reduce the levels of PTGS2 and ACSL4. Moreover, berberine significantly improved the levels of ROS and GSH in podocytes under HG conditions, thereby alleviating membrane blistering, mitochondrial shrinkage and other morphological changes of HG-induced podocytes. Conclusions In this study, the number of podocytes decreases, which is a death mode different from autophagy and apoptosis, that is, ferroptosis. Berberine can alleviate the occurrence of this phenomenon by mediating the Nrf2/ H0-1/GPX4.