MMP13 Antibody - #DF6494

Fig. 2. Quercitrin suppressed MMP13 expression and increased collagen Ⅱ deposition in IL-1b-induced rat chondrocytes and SW1353 cells. (A–D) Rat chondrocytes and SW1353 cells were pretreated with IL-1b (10 ng/ml) for 2 h prior to treatment with different concentrations of quercitrin (12.5, 25, and 50 lM) for 48 h. The cells were collected and collagen II and MMP13 mRNA expression levels were evaluated by RT-PCR. (E-J) Rat chondrocytes and SW1353 cells were pre-treated with IL-1b (10 ng/ml) for 2 h prior to treatment with different concentrations of quercitrin (12.5, 25, and 50 lM) for 48 h. The cells were collected and collagen II and MMP13 protein expression levels were evaluated by western blot. RT-PCR and western blot were repeated at least three times with similar results. All data are represented as the mean ± SD. *P < 0.05, **P < 0.01and ***P < 0.001 versus the control group; #P < 0.05, ##P < 0.01 and ###P < 0.001 versus the IL-1b-treated group. (K) Compared with control (n = 6), p110a mRNA expression levels were significantly decreased in severe OA patients (n = 12) by RT-PCR. ***P < 0.001 versus the control. Data from RT-PCR and western blot were analyzed by using one-way ANOVA, followed by Tukey’s range test.

Fig. 2. Effects of avicularin on ECM degradation. (A-D) The mRNA levels of ACAN, Collagen 2, MMP3 and MMP13 in rat and human chondrocytes were detected by qPCR. The dates were shown as the means ± S.D. ** p < 0.01 and *** p < 0.001 compared with the control group; # p < 0.05, ## p < 0.01 and ### p < 0.001 compared with the IL-1β-induced group, n = 3. (E-F) The expression of MMP3, MMP13, ADAMTS5, SOX9, ACAN and Collagen II was detected by western blots.

Fig. 7. Avicularin attenuates ECM degradation in ACLT rats. (A) ACAN, (B) Collagen II, (C) MMP3, (D) MMP13 and (E) TRAF6 immunohistochemistry of knee joint medial compartment cartilage. (F-J) Quantitative analysis of ACAN, Collagen II, MMP3, MMP13 and TRAF6. The values were shown as the means ± S.D. ** p < 0.01 and *** p < 0.001 compared with the control group; # p < 0.05, ## p < 0.01 and ### p < 0.001 compared with the ACLT group, n = 6. (K) The working model of how avicularin protects against OA development.

Figure 3. Key indicators expression of knee cartilage in 7-day male mice offspring of control and obese groups. (A) Relative mRNA, (B) representative western blot bands, and (C) statistical results of knee cartilage 7-day male mice offspring in the control group versus obese group. O-WT-P7: offspring 7-day male mice from the control mothers. O-OB-P7: offspring 7-day male mice from the obese mothers. (A) n = 6 represents six mice in each group from at least three litters of different mothers. (B) n = 6, each band represents the expression of cartilage tissue proteins in two mice from at least three litters of different mothers. Results are expressed as mean ± SD. ns not significant, * p < 0.05, ** p < 0.01, and *** p < 0.001.

Fig. 6. (A) Analysis of the cytotoxicity induced by the 6 lignans in SW1353 cells. (B) Effects of the 6 lignans on collagen II gene expression in IL-1β- treated SW1353 cells. (C) Effects of the 6 lignans on MMP13 gene expression in IL-1β-treated SW1353 cells. (D) Effects of schisanhenol and gammaschisandrin on collagen II gene expression in IL-1β- treated SW1353 cells. (E) Effect of schisanhenol on MMP13 and collagen II protein expression in IL-1β- treated SW1353 cells. All the data are represented as the mean ± SD. Ns represents not significant, *p < 0.05, **p < 0.01 and ***p < 0.001, versus the control group; #p < 0.05, ##p < 0.01 and ###p < 0.001, versus the IL-1β-treated group.

Figure 2 Effect of irisin on MMP-13 and Col-II in IL-1β-induced SW1353 cells. SW1353 cells were treated with 20 mM irisin for 24 h, with or without 10 ng/ml IL-1β. (A) Col II protein levels, (B) MMP-13 protein levels, (C) Col II and MMP-13 protein expression, (D) Col II mRNA levels and (E) MMP-13 mRNA levels were evaluated by reverse transcription-quantitative PCR and western blot analysis (n=5/group). *P<0.05, **P<0.01 and ***P<0.001. IL-1β, interleukin-1β; MMP-13, matrix metalloproteinase-13; Col-II, collagen II; NC, negative control.