HSP60 Regulates Monosodium Urate Crystal-Induced Inflammation by Activating the TLR4-NF-κB-MyD88 Signaling Pathway and Disrupting Mitochondrial Function

Figure 5

HSP60 knockdown alleviates mitochondrial dysfunction in MSU crystal-stimulated THP-1-derived macrophages. (a–g) THP-1-derived macrophages were transfected with control siRNA or HSP60 siRNA for 48 h, primed with LPS (100 ng/ml) for 1 h, and then treated with MSU suspension (50 μg/ml) for 12 h. (a) The effect of HSP60 knockdown on the mitochondrial ROS, representative images of MitoTracker green and MitoROS staining. Blue shows nuclei staining with Hoechst33342. Scale bar: 20 μm. (b) SOD activity. (c) CAT activity. (d) GSH-Px activity. (e) Mitochondrial DNA release was detected by quantitative real-time PCR analysis. (f) Relative mitochondrial ATP content. (g) The effect of HSP60 knockdown on the mitochondrial membrane potential (MMP), cells were stained using JC-1 probe. Blue shows nuclei staining with Hoechst33342. Scale bar: 20 μm. For mtROS and MMP analysis, >50 individual cells were imaged per group from 3 culture dishes. Data are representative of for three experiments. Each experiment had 10 fields of view. .

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