FIGURE 2.

In vitro cardioprotective activity of puerarin against hypoxia/reoxygenation injury. (A) Determination of LDH release from H9c2 cells. After exposure to 3 h hypoxia (0.1% O₂) and 24 h reoxygenation, H9c2 cells were treated with/without puerarin (10, 20, and 40 μM) for 1 h. LDH release was measured by a commercial LDH cytotoxicity assay kit. (B) Western blot analysis of COX2, galectin-3 and cleaved-PARP-1 expression. H9c2 cells were collected and the proteins were analyzed by Western blot with antibodies against COX2, galectin-3, cleaved-PARP-1. (C) Quantification of COX2, galectin-3 and cleaved-PARP-1 expression. The blots from Panel B were quantified by a densitometric method. (D) Immunostaining of 8-OHdG. H9c2 cells were probed with anti-8-OHdG antibody and visualized with Alexa Fluor 488-conjugated secondary antibody, whereas the cell nuclei were stained with DAPI. Scale bar, 50 μm. (E) Quantification of 8-OHdG generation. The results were presented as mean ± SD (n = 3). *p < 0.05.