Research Paper Volume 12, Issue 18 pp 18588—18602

Lipopolysaccharide upregulates miR-132/212 in Hirschsprung-associated enterocolitis, facilitating pyroptosis by activating NLRP3 inflammasome via targeting Sirtuin 1 (SIRT1)

Figure 4. Activation of NLRP3 inflammasome in patients with postoperative HAEC and LPS-stimulated HT29 cell line. (A) qRT-PCR validated relative mRNA levels of NLRP3, Caspase-1 and ASC in dilated segments of 32 po-HAEC and 32 No-HAEC patients. The above genes mRNA expression was distinctly increased in po-HAEC tissues. Compared with control mice tissues, NLRP3, Caspase-1 and ASC mRNA levels showed a increase in mice intestinal tissues challenged with LPS. (B) The protein levels of NLRP3, Caspase-1 and ASC were up-regulated in 4 pairs of po-HAEC tissues compared to No-HAEC samples. LPS treatment to mice increased NLRP3, Caspase-1 and ASC protein expression of intestinal tissues. β-actin was regarded as an internal control. (C and D) The mRNA and protein levels of NLRP3, Caspase-1 and ASC were upregulated in HT29 cell line after treatment with LPS for 24h. Mean ± SD. n = 3, *P < 0.05. (E) Confocal images of SIRT1 and NLRP3 in HT29 cell line treated with 1000 ng/ml LPS for 24 h. Scale bar =25μm.