FIGURE 4.

Neferine suppressed apoptosis in LPS-treated H9c2 cells (A) Cell Counting Kit-8 Assay (CCK8) was performed to assess cell viability in H9c2 cells that were treated with a concentration gradient of LPS (5, 10, 20, and 50 μg/ml) for 24 h (B) The cytotoxicity of different neferine concentrations (0.25, 0.5, 1, 2, 4, 8, 16, 32, and 64 μM) in H9c2 cells was detected by CCK8 assay (C) Pretreatment with neferine (0.25, 0.5, 1, 2, 4, and 8 μM) for 2 h prior to LPS exposure (10 μg/ml) for 12 h; cell viability was tested by CCK8 assay (D–F) Western blot analysis and densitometric quantification of the protein expression levels of cleaved caspase 3 and Bcl-2 (G) TUNEL staining-positive cells (green) labeled apoptotic cells, DAPI (blue) labeled H9c2 cells nuclei. Scale bar, 100 μm (H) Measurement of TUNEL-positive cell ratio. All data are expressed as mean ± standard deviation. All experiments were repeated at least three times. *p < 0.05, **p < 0.01, ***p < 0.001; ns: no significant difference; TUNEL, terminal deoxynucleotidyl transferase dUTP nick end labeling; DAPI, 4′,6-diamidino-2-phenylindole.