Fig 1: Intramyocardial injection of human extracellular vesicles (EVs) reduce activation of the nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-3 (NLRP3) inflammasome in a model of rat sterile pericarditis. (A) Schematic outline of the inflammatory effect of post operative pericarditis on NLRP3 activation, atrial remodeling and EV effects (red boxes). (B) Effect of pericarditis and EV treatment on the atrial content of NLRP3 and downstream mediators (interleukin-1 beta (IL-1β) and IL-18). NLRP3 data was analyzed using a one-way ANOVA with individual-mean comparisons by Šídák's multiple comparisons test. *P<0.05 vs. talc + vehicle treated animals. Relationship between NLRP3 and IL-1β or IL-18 abundance within atrial tissue was analyzed using simple linear regression. (C) Rank order plot highlighting the abundance of miRNAs (left panel) and proteins (right panel) found in EVs suggested to directly inhibit NLRP3 activation. (D) In vitro effect of explant-derived cell (EDC) EVs on caspase-1 activity, a measure of NLRP3 activation, in THP-1 cells (macrophages) and primary cultured atrial fibroblasts after activation by lipopolysaccharide (LPS) and nigericin. One-way ANOVA with individual-mean comparisons by Šídák's multiple comparisons test. *P<0.05 vs. untreated cells; #P<0.05 vs. LPS+nigericin treated cells. (E) In vitro effect of EDC EVs on downstream mediators of NLRP3 activation (IL-1β and IL-18) in THP-1 cells (macrophages) after activation by LPS and nigericin. One-way ANOVA with individual-mean comparisons by Šídák's multiple comparisons test. *P<0.05 vs. untreated cells; #P<0.05 vs. LPS+nigericin treated cells. (F) Comparative effect of EVs from bone marrow mesenchymal stromal cells (BM-MSCs), umbilical cord mesenchymal stromal cells (UC-MSCs) or EDCs on caspase-1 activity, the principal effector of NLRP3 activation. One-way ANOVA with individual-mean comparisons by Šídák's multiple comparisons test. *P<0.05 vs. untreated cells; #P<0.05 vs. BM-MSC or UC-MSC treated cells. NEDD4, neural precursor cell expressed developmentally down-regulated protein 4; PRDX1, Peroxiredoxin-1; SQSTM1, Sequestosome 1.
Fig 2: Molecular mechanism of BMP7 involved in the IDD of T1DM rats. BMP7 inhibits NPC pyroptosis and NLRP3 inflammasome activity to ameliorate IDD in T1DM
Fig 3: Effect of the different doses of canagliflozin on the hippocampal TXNIP/NF-κB p65/NLRP3 inflammasome signalling in rats injected with aluminium chloride (mean ± SD); *** = p < 0.001. AlCl3, aluminium chloride; CNG, canagliflozin.
Fig 4: BMP7 affects the IDD of T1DM rats by regulating NLRP3 inflammasome activity and NPC pyroptosis. STZ-induced T1DM rats were treated with oe-BMP7 alone or combined with 4’MR (n = 12). A Measurement of disc height of STZ-induced T1DM rats. B HE staining of NP tissues from STZ-induced T1DM rats. C Safranin O-fast green staining for NP tissues and AF tissues of STZ-induced T1DM rats. D ECM-related gene expression in NP tissues from STZ-induced T1DM rats. E BMP7 expression in NP tissues of STZ-induced T1DM rats determined by RT-qPCR. F BMP7 levels in NP tissues of STZ-induced T1DM rats measured by ELISA. G Levels of pyroptosis-related proteins in NP tissues of STZ-induced T1DM rats measured by ELISA. H Apoptosis in NP tissues of STZ-induced T1DM rats detected by TUNEL staining. I Ki67-positive cells in NP tissues of STZ-induced T1DM rats detected by IHC. *p < 0.05. Data are shown as the mean ± standard deviation. Data comparisons among multiple groups were analyzed by one-way ANOVA with Tukey’s post hoc test
Fig 5: Effect of BMP7 on the IDD of T1DM rats by regulating NLRP3 inflammasome activity and pyroptosis of NPCs. A BMP7 expression in the transcriptome sequencing data of T1DM-induced IDD rats. Control (normal rats) n = 3, STZ (T1DM-induced IDD rats) n = 3. B BMP7 expression in the NP tissues of STZ-induced T1DM rats measured by RT-qPCR. C BMP7 expression in the NPCs isolated from STZ-induced T1DM rats measured by RT-qPCR. D Levels of pyroptosis-related proteins in NPCs and cell supernatant measured by ELISA. E BMP7 expression in NPCs from STZ-induced T1DM rats transduced with oe-BMP7 and treated with 4’MR determined by RT-qPCR. F BMP7 levels in NPCs and cell supernatant from STZ-induced T1DM rats transduced with oe-BMP7 and treated with 4’MR measured by ELISA. G Levels of pyroptosis-related proteins in NPCs from STZ-induced T1DM rats transduced with oe-BMP7 and treated with 4’MR measured by ELISA. n = 12. *p < 0.05. n.s. indicates not significant. Data are shown as the mean ± standard deviation. Data between two groups were compared by unpaired t-test, and data comparisons among multiple groups were analyzed by the one-way ANOVA with Tukey’s post hoc test. The cell experiment was repeated three times independently
Supplier Page from Abcam for Rat NLRP3 ELISA Kit