Fig. 6.

Blockade effects of the specific α7-nicotinic acetylcholine receptor (α7-nAChR) antagonist methyllycaconitine on cinobufagin (a, b)- and the specific α7-nAChR agonist PHA-543613 (c, d)-stimulated gene expression of IL-10 and the β-endorphin precursor proopiomelanocortin (POMC) in primary cultures of spinal microglia. e–h. Effects of PHA-543613 and ouabain on the protein expression of IL-10 and β-endorphin. Microglial cells, originated from the spinal cords of 1-day-old neonatal rats, were incubated with cinobufagin (100 μM), PHA-543613 (100 μM), or ouabain (10 μM). The mRNA and protein expression of IL-10 and POMC was measured 2 h later using qRT-PCR and the commercial kits, respectively. For the methyllycaconitine blockade study, methyllycaconitine was incubated 0.5 h prior to cinobufagin or PHA-543613 treatment. 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 and cinobufagin group, respectively, by one-way ANOVA followed by the post hoc Student–Newman–Keuls test