Fig. 3.

Effects of cinobufagin on the gene (a–d) and protein (e, f) expression of IL-10, the β-endorphin precursor proopiomelanocortin (POMC) and dynorphin precursor prodynorphin (PDYN) in the primary cultures of spinal microglia, neurons, and astrocytes. g, h The blockade effect of the microglial inhibitor minocycline on cinobufagin-induced gene overexpression of IL-10 and POMC. The primary culture cells, originated from the spinal cords of 1-day-old neonatal rats, were collected 2 h after cinobufagin incubation. For the blockade study, minocycline was incubated 1 h prior to cinobufagin treatment. The mRNA expression of IL-10, POMC, and PDYN was measured by using qRT-PCR. The data are presented as means ± SEM (n = 3 independent repeats with duplicates). The asterisk and number sign denote statistical significance (p < 0.001) compared to the control group and cinobufagin group, respectively, by unpaired and two-tailed Student t test (a–f) or one-way ANOVA followed by the post hoc Student–Newman–Keuls test (g, h)