Isoliquiritigenin Inhibits IL-1β-Induced Production of Matrix Metalloproteinase in Articular Chondrocytes Lei Zhang, Shiyun Ma, Hang Su, Jiaxiang Cheng

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Isoliquiritigenin Inhibits IL-1β-Induced Production of Matrix Metalloproteinase in Articular Chondrocytes Lei Zhang, Shiyun Ma, Hang Su, Jiaxiang Cheng Molecular Therapy - Methods & Clinical Development Volume 9, Pages 153-159 (June 2018) DOI: 10.1016/j.omtm.2018.02.006 Copyright © 2018 The Authors Terms and Conditions

Figure 1 Effect of Isoliquiritigenin on Proliferation of Rabbit Chondrocytes Chondrocytes were incubated for 72 hr in the presence of varying concentrations of isoliquiritigenin. Cell viability was determined using SRB assay as described in Materials and Methods. Each bar represents a mean ± SEM of three independent experiments in comparison with that of the control set at 100%. Molecular Therapy - Methods & Clinical Development 2018 9, 153-159DOI: (10.1016/j.omtm.2018.02.006) Copyright © 2018 The Authors Terms and Conditions

Figure 2 Effect of Isoliquiritigenin on the Gene Expression of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, and ADAMTS-5 in Chondrocytes Primary cultured articular chondrocytes were pretreated with varying concentrations of isoliquiritigenin for 2 hr and then stimulated with IL-1β (10 ng/mL) for 24 hr. The gene expression levels of MMP-1 (A), MMP-3 (B), MMP-9 (C), MMP-13 (D), ADAMTS-4 (E), and ADAMTS-5 (F) were measured by real-time PCR. Three independent experiments were performed and the representative data were shown. Each bar represents a mean ± SEM of three independent experiments in comparison with that of the control set at 100%. *Significantly different from control (p < 0.05). Significantly different from IL-1β alone (#p < 0.05). Molecular Therapy - Methods & Clinical Development 2018 9, 153-159DOI: (10.1016/j.omtm.2018.02.006) Copyright © 2018 The Authors Terms and Conditions

Figure 3 Effect of Isoliquiritigenin on the Protein Levels of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, and ADAMTS-5 in Chondrocytes Primary cultured articular chondrocytes were pretreated with varying concentrations of isoliquiritigenin for 2 hr and then stimulated with IL-1β (10 ng/mL) for 24 hr. The protein levels of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, and ADAMTS-5 were measured by western blotting (A). Relative protein expressions to control were shown as histogram in (B). Six independent experiments were performed and the representative data were shown. Each bar represents a mean ± SEM of three independent experiments in comparison with that of the control set at 100%. Significantly different from control (*p < 0.05). Significantly different from IL-1β alone (#p < 0.05). Molecular Therapy - Methods & Clinical Development 2018 9, 153-159DOI: (10.1016/j.omtm.2018.02.006) Copyright © 2018 The Authors Terms and Conditions

Figure 4 Effect of Isoliquiritigenin on Phosphorylation of NF-κB and IκBα in Chondrocytes Primary cultured articular chondrocytes were pretreated with varying concentrations of isoliquiritigenin for 2 hr and then stimulated with IL-1β (10 ng/mL) for 24 hr. The phosphorylation of NF-κB and IκBα were measured by western blotting (A). Relative protein expressions to control were shown as histogram in (B). Six independent experiments were performed and the representative data were shown. Each bar represents a mean ± SEM of three independent experiments in comparison with that of the control set at 100%. Significantly different from control (*p < 0.05). Significantly different from IL-1β alone (#p < 0.05). Molecular Therapy - Methods & Clinical Development 2018 9, 153-159DOI: (10.1016/j.omtm.2018.02.006) Copyright © 2018 The Authors Terms and Conditions

Figure 5 Effect of Isoliquiritigenin on the Protein Levels of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, ADAMTS-5 and the Phosphorylation of NF-κB and IκBα In Vivo The knee joint of rats was pretreated with 20, 50, or 100 mg/kg of isoliquiritigenin for 3 hr and then stimulated with IL-1β (20 ng/20 μL) for 72 hr by intra-articular injection. Tissue lysates from articular cartilage homogenates were collected for measuring the protein levels of MMP-1, MMP-3, MMP-9, MMP-13, ADAMTS-4, and ADAMTS-5 (A) and the phosphorylation of NF-κB and IκBα (C) in vivo by western blot analysis. Relative protein expressions to control were shown in (B) and (D), respectively. Six independent experiments were performed and the representative data were shown. Each bar represents a mean ± SEM of three independent experiments in comparison with that of the control set at 100%. Significantly different from control (*p < 0.05). Significantly different from IL-1β alone (#p < 0.05). Molecular Therapy - Methods & Clinical Development 2018 9, 153-159DOI: (10.1016/j.omtm.2018.02.006) Copyright © 2018 The Authors Terms and Conditions