NLRP3 inflammasome expression in peripheral blood monocytes of coronary heart disease patients and its modulation by rosuvastatin

Figures

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  Figure 1.. Upregulation of NLRP3 inflammasome mRNA levels in PBMCs in patients with SAP and AMI, compared with non-CHD controls. (A) PBMCs were isolated from peripheral blood and the positive rate of CD14 was calculated to be ≥95% of PBMCs by flow cytometry. RT-PCR assays were performed to quantify the mRNA levels of (B) NLRP3, (C) ASC and (D) caspase-1 in PBMCs of each group. *P<0.05; **P<0.01. NLRP3, nucleotide-binding oligomerization domain, leucine rich repeat, and pyrin domain-containing protein 3; PBMCs, peripheral blood monocytes; SAP, stable angina pectoris; AMI, acute myocardial infarction; CHD, coronary heart disease; ASC, apoptosis-associated speck-like protein containing a CARD.

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  Figure 2.. Increase in protein expression levels of NLRP3 inflammasome and secretion of its downstream interleukins in PBMCs of patients with SAP and AMI, compared with non-CHD controls. (A) Western blotting was used to detect the protein expression of NLRP3, ASC and caspase-1 in PBMCs of each group. (B) Quantitative results are illustrated for the three indicators using western blot analysis. (C) Plasma levels of IL-1β and IL-18 from each group were measured by ELISA. *P<0.05; **P<0.01. NLRP3, nucleotide-binding oligomerization domain, leucine rich repeat, and pyrin domain-containing protein 3; PBMCs, peripheral blood monocytes; SAP, stable angina pectoris; AMI, acute myocardial infarction; CHD, coronary heart disease; ASC, apoptosis-associated speck-like protein containing a CARD; ELISA, enzyme-linked immunosorbent assay.

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  Figure 3.. Effect of rosuvastatin on NLRP3 inflammasome mRNA levels in PBMCs in vitro. PBMCs were isolated from the peripheral blood of patients with non-CHD, SAP and AMI, respectively, followed by proliferation and treatment with rosuvastatin at a concentration of 20 µM for the indicated time-points. RT-PCR assay was performed to investigate the influence of rosuvastatin on the mRNA levels of (A) NLRP3, (B) ASC and (C) caspase-1 in PBMCs of each group. *P<0.05; **P<0.01. NLRP3, nucleotide-binding oligomerization domain, leucine rich repeat, and pyrin domain-containing protein 3; PBMCs, peripheral blood monocytes; CHD, coronary heart disease; SAP, stable angina pectoris; AMI, acute myocardial infarction; ASC, apoptosis-associated speck-like protein containing a CARD; NS, not significant.

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  Figure 4.. Effect of rosuvastatin on protein expression levels of NLRP3 inflammasome in PBMCs and secretion of its downstream cytokines in supernatants in vitro. PBMCs were isolated from the peripheral blood of patients with non-CHD, SAP and AMI, respectively, followed by proliferation and treatment with rosuvastatin at a concentration of 20 µM for the indicated time-points. (A) Western blotting was conducted to analyze the influence of rosuvastatin on the protein levels of NLRP3, ASC and caspase-1 in PBMCs of each group. (B) Quantitative results for modulating the protein expression of the aforementioned three indicators by rosuvastatin are illustrated. (C) Modulation of IL-1β and IL-18 secretion in the supernatants by rosuvastatin of each group was measured by ELISA. *P<0.05, **P<0.01 and #P<0.05 (compared to the 0-h group); ΔP<0.05 (compared to the 12-h group). NLRP3, nucleotide-binding oligomerization domain, leucine rich repeat, and pyrin domain-containing protein 3; PBMCs, peripheral blood monocytes; CHD, coronary heart disease; SAP, stable angina pectoris; AMI, acute myocardial infarction; ASC, apoptosis-associated speck-like protein containing a CARD; NS, not significant.

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  Figure 5.. Proposed model for NLRP3 inflammasome signaling pathway involved in inflammatory pathogenesis of coronary atherosclerosis. Stimulated PBMCs orchestrate activation of NLRP3 inflammasome due to the upregulation of the expression of NLRP3, ASC, caspase-1, and their downstream mediators, leading to atherosclerotic progression. NLRP3, nucleotide-binding oligomerization domain, leucine rich repeat, and pyrin domain-containing protein 3; PBMCs, peripheral blood monocytes; ASC, apoptosis-associated speck-like protein containing a CARD.