Naofen promotes TNF-α-mediated apoptosis of hepatocytes by activating caspase-3 in lipopolysaccharide-treated rats

Figure 4 Effect of Kupffer cell-conditioned medium on naofen mRNA expression and caspase-3 activation. Primary Kupffer cell (KCs) and hepatocytes were separated as described above, and KC-CM was obtained by incubating KCs with LPS (100 ng/mL) for 6 h. Anti-TNF-α antibody (500 ng/mL) was added to KC-CM treated with LPS for 6 h and incubated at 37 °C for 1 h (anti-TNF-α + 6 h KC-CM). Hepatocytes were incubated with LPS (100 ng/mL), TNF-α (10 ng/mL), IgG (500 ng/mL), 6 h KC-CM and anti-TNF-α + 6 h KC-CM for 12 h, respectively. A: Naofen mRNA in hepatocytes was measured with qPCR and GAPDH was used as an internal control (n = 6); B: Immunoblotting assay for naofen. Hepatocyte lysates were analyzed with naofen antibody (n = 6); C: Caspase-3 activation was also measured (n = 6). ^bP < 0.01 vs controls; ^cP < 0.05 and ^dP < 0.01 vs 6 h KC-CM. TNF-α: Tumor necrosis factor-α; LPS: Lipopolysaccharide; KC-CM: Kupffer cell-conditioned medium.