Fig 2: MATN3 suppresses IL-1β-induced ECM degradation and autophagy defects in chondrocytes via the IL-17A/PI3K/AKT/mTOR axis. A Western blot for the protein levels of PI3K, AKT, and mTOR and phosphorylation levels of PI3K, AKT, and mTOR in cells. B, CCK-8 for the viability of chondrocytes. C-D, ELISA for IL-6 and TNF-α contents in the supernatants of chondrocytes. E, Western blot for the expression levels of MMP-13, ADAMTS5, aggrecan, Collagen II, LC3II/LC3I, and Beclin1 in chondrocytes. ∗ indicates p ​< ​0.05 compared with control/DMSO group; # indicates p ​< ​0.05 compared with the IL-1β + sh-NC/IL-1β + sh-IL-17A ​+ ​DMSO group; & indicates p ​< ​0.05 compared with the IL-1β ​+ ​oe-NC/IL-1β ​+ ​oe-MATN3 ​+ ​DMSO group. Cell experiments were performed in triplicate.

Fig 3: MATN3 inhibits IL-1β-induced ECM degradation and autophagy defects in chondrocytes through interaction with IL-17A. A, RT-qPCR for IL-17A expression in chondrocytes. B, Western blot for IL-17A and MATN3 protein levels in chondrocytes. C, RT-qPCR for mRNA levels of IL-17A in the cartilage tissues of mice. D, Western blot for IL-17A and MATN3 protein levels in the cartilage tissue of mice. E, Co-IP experiment for the interaction between IL-17A and MATN3. F, Immunofluorescence for the co-localization of IL-17 and MATN3 in chondrocytes (bar ​= ​25 ​μm). G-H, ELISA for the IL-6 and TNF-α content in the supernatants of the chondrocytes. I, Western blot for expression of MMP-13, ADAMTS5, aggrecan, Collagen II, LC3II/LC3I, and Beclin1 in chondrocytes. ∗ indicates p ​< ​0.05 compared with the control group/sham; # indicates p ​< ​0.05 compared with the IL-17A ​+ ​oe-NC/OA ​+ ​oe-NC group; & indicates p ​< ​0.05 compared with the IL-17A ​+ ​sh-NC group. Cell experiments were performed in triplicate.

Fig 4: The mechanism diagram. The mechanism diagram showed that dsHMGB1 released from IL-17A induced pyrosis of prostatic epithelial cells, which affected Jak2/Stat1 transcriptional regulation of Pfkp, led to changes in macrophage glucose metabolism, and further affected M1 polarization process (By Figdraw).

Fig 5: dsHMGB1 released from IL-17A-induced pyroptotic prostate epithelial cells. (A-D) The flow chart, HE staining, inflammation score and pain response frequency in control, EAP and EAP+anti-IL-17A groups of mice; (E) The RNA levels of inflammatory factors IL-1β, IL-6 and TNF-α between control, EAP and EAP+ anti-IL-17A groups were detected by RT-qPCR; (F-G) The TUNEL and GSDMD staining between control, EAP and EAP+anti-IL-17A group; (H) The levels of IL-17A and dsHMGB1 among the control, EAP and EAP+anti-IL-17A groups was detected by ELISA; (I-J) The expression levels of HMGB1, GSDMD-NT, GSDMD, IL-18, Cleaved IL-1β, NLRP3, ASC, Pro Caspase-1 and Cleaved Caspase-1 among the control, EAP and EAP+anti-IL-17A groups was detected by western blot; (K) The immunofluorescence staining of HMGB1 among the control, EAP and EAP+anti-IL-17A groups; (L-M) The expression levels of HMGB1, GSDMD-NT, GSDMD, IL-18, Cleaved IL-1β, NLRP3, ASC, Pro Caspase-1 and Cleaved Caspase-1 among the control and IL-17A groups was detected by western blot; (N) The RNA levels of HMGB1 between control and IL-17A groups were detected by RT-qPCR; (O) The levels of dsHMGB1 between control and IL-17A groups were detected by ELISA; (P) The co-immunofluorescence staining of Caspase-1 and GSDMD between control and IL-17A groups. *P < 0.05; **P < 0.01; ***P < 0.001.