Fig. 8

The effects of SAM and NaHS administration on NLRP3 inflammasome activation in primary microglial cells. a Quantitative analysis of NLRP3 mRNA levels by qPCR in the control, LPS + ATP, LPS + ATP + SAM and LPS + ATP + NaHS groups. The relative densities of each mRNA have been normalized against those of the LPS + ATP group. LPS and ATP stimulation significantly enhanced NLRP3 mRNA levels (p < 0.01 vs. control). Both SAM and NaHS treatment inhibited this enhancement (p < 0.05 vs. LPS + ATP). b–f Representative bands and quantitative analysis of NLRP3 and ASC expression in the microglial cell lysates and caspase-1 p20 subunit and mature IL-1β levels in the medium and in the control, LPS + ATP, LPS + ATP + SAM and LPS + ATP + NaHS groups. Compared with the control group, LPS and ATP stimulation increased the protein levels of the NLRP3 inflammasome components in the LPS + ATP group, leading to increased levels of IL-1β and caspase-1 release in the medium (p < 0.01), but SAM and NaHS treatment significantly suppressed these upregulations (p < 0.05). The relative densities of each protein have been normalized against those of the LPS + ATP group. N = 3 per group, Data are presented as the mean ± SEM. &&p < 0.01 vs. control; @p < 0.05 vs. LPS + ATP. SAM S-adenosyl-l-methionine, GAPDH glyceraldehyde 3-phosphate dehydrogenase, IL interleukin, NLRP3 pyrin domain-containing 3, ASC adaptor protein apoptosis-associated speck-like protein containing a CARD