Journal: Annals of Translational Medicine

Article Title: MicroRNA-25-3p therapy for intervertebral disc degeneration by targeting the IL-1β/ZIP8/MTF1 signaling pathway with a novel thermo-responsive vector

doi: 10.21037/atm-20-6595

Figure Lengend Snippet: The influence of IL-1β on ECM-degrading enzyme (MMP3, MMP13 and ADAMTS5) and ECM protein (collagen type II and aggrecan) expression. (A) Relative expression levels of MMP3 mRNA, MMP13 mRNA and ADAMTS5 mRNA in IL-1β-stimulated nucleus pulposus (NP) cells (10 ng/mL) and control cells via qRT-PCR. (B) Relative protein expression levels of MMP3, MMP13 and ADAMTS5 were examined in IL-1β-stimulated NP cells (10 ng/mL) and control cells via western blotting. (C) Relative protein expression levels of collagen type II and aggrecan were detected in IL-1β-stimulated NP cells (10 ng/mL) and control cells via immunocytochemistry (ICC). The bar is 50 µm. Data are the mean ± standard deviation (SD) of triplicate results. *, P<0.05.

Article Snippet: Membranes were incubated overnight at 4 °C with anti-IL-1β antibody (12703, 1:1,000, CST, USA), anti-MTF1 antibody (ab184119, 1:1,000, Abcam, Cambridge, UK), anti-ZIP8 antibody (NBP1-76505, 1:2,000, Novus, Biologicals, Colorado, USA), anti-MMP3 antibody (NB100-91878, 1:1,000, Novus Biologicals, Colorado, USA), anti-MMP13 antibody (NBP2-45887, 1:1,000, Novus, Biologicals, Colorado, USA), anti-ADAMTS5 antibody (NBP2-15286, 1:1,000, Novus, Biologicals, Colorado,USA), anti-β-actin antibody (abs118937, 1:3,000, Absin, Shanghai, China), and anti-PCNA antibody (abs13075, 1:2,000, Absin, Shanghai, China).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Immunocytochemistry, Standard Deviation

Journal: Annals of Translational Medicine

Article Title: MicroRNA-25-3p therapy for intervertebral disc degeneration by targeting the IL-1β/ZIP8/MTF1 signaling pathway with a novel thermo-responsive vector

doi: 10.21037/atm-20-6595

Figure Lengend Snippet: The effects of miRNA-25-3p modulation on IL-1β-stimulated ECM-degrading enzyme (MMP3, MMP13 and ADAMTS5) and ECM protein (collagen type II and aggrecan) expression. (A) Relative mRNA expression levels of MMP3, MMP13 and ADAMTS5 in the miRNA-25-3p mimic group and the control group determined via qRT-PCR. (B) Relative protein expression levels of MMP3, MMP13 and ADAMTS5 were examined in the miRNA-25-3p mimic group and the control group via western blotting. (C) Relative protein expression levels of collagen type II and aggrecan were detected in the miRNA-25-3p mimic group and the control group via immunocytochemistry (ICC). The bar is 50 µm. Data are the mean ± standard deviation (SD) of triplicate results. *, P<0.05.

Article Snippet: Membranes were incubated overnight at 4 °C with anti-IL-1β antibody (12703, 1:1,000, CST, USA), anti-MTF1 antibody (ab184119, 1:1,000, Abcam, Cambridge, UK), anti-ZIP8 antibody (NBP1-76505, 1:2,000, Novus, Biologicals, Colorado, USA), anti-MMP3 antibody (NB100-91878, 1:1,000, Novus Biologicals, Colorado, USA), anti-MMP13 antibody (NBP2-45887, 1:1,000, Novus, Biologicals, Colorado, USA), anti-ADAMTS5 antibody (NBP2-15286, 1:1,000, Novus, Biologicals, Colorado,USA), anti-β-actin antibody (abs118937, 1:3,000, Absin, Shanghai, China), and anti-PCNA antibody (abs13075, 1:2,000, Absin, Shanghai, China).

Techniques: Expressing, Control, Quantitative RT-PCR, Western Blot, Immunocytochemistry, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Detection of miR-29a in plasma of patients with lumbar spinal stenosis and the clinical significance

doi: 10.3892/mmr.2018.8956

Figure Lengend Snippet: Comparison of plasma levels of (A) MMP9 and (B) ADAMTS5 between different patient groups. Triangles and black dots represent the ROC curve of LDH and LSS, respectively. The gray lines are lines of identity. **P<0.01, compared with the control group. ## P<0.01, compared with patients with LDH. ADAMTS5; a disintegrin and metalloproteinase with thrombospondin motifs 5; control, healthy patients; LDH, lumbar intervertebral disc herniation; LSS, lumbar spinal stenosis; MMP9, matrix metalloproteinase 9.

Article Snippet: Human MMP-9 Quantikine ELISA kit DMP900 (R&D Systems, Inc., Minneapolis, MN, USA) and Human ADAMTS5 DuoSet ELISA kit (R&D Systems, Inc.) were used according to the manufacture's protocols.

Techniques: Comparison, Clinical Proteomics, Control

Journal: Molecular Medicine Reports

Article Title: Detection of miR-29a in plasma of patients with lumbar spinal stenosis and the clinical significance

doi: 10.3892/mmr.2018.8956

Figure Lengend Snippet: Comparison of MMP9 and ADAMTS5 expression levels in intervertebral disc tissue of various patient groups. (A) Expression levels of MMP9 and ADAMTS5 in intervertebral disc tissue of various patient groups; (B) Expression levels of ADAMTS5 in intervertebral disc tissue of various patient groups. **P<0.01, compared with the control group. ## P<0.01, compared with patients with LDH. ADAMTS5; a disintegrin and metalloproteinase with thrombospondin motifs 5; control, healthy patients; LDH, lumbar intervertebral disc herniation; LSS, lumbar spinal stenosis; MMP9, matrix metalloproteinase 9.

Article Snippet: Human MMP-9 Quantikine ELISA kit DMP900 (R&D Systems, Inc., Minneapolis, MN, USA) and Human ADAMTS5 DuoSet ELISA kit (R&D Systems, Inc.) were used according to the manufacture's protocols.

Techniques: Comparison, Expressing, Control

Journal: Molecular Medicine Reports

Article Title: Detection of miR-29a in plasma of patients with lumbar spinal stenosis and the clinical significance

doi: 10.3892/mmr.2018.8956

Figure Lengend Snippet: Correlations between the expression levels of miR-29, MMP9 and ADAMTS5, and correlation between their expression levels in plasma and intervertebral disc tissue of LSS patients. (A) Correlation between the expression levels of miR-29 and MMP9. (B) Correlation between the expression levels of miR-29 and ADAMTS5. (C) Correlation between the expression levels of miR-29 in plasma and intervertebral disc tissue. Correlation between the expression levels of (D) MMP9 and (E) ADAMTS5 in plasma and intervertebral disc tissue. y, the binary equation representing the distributions of values; ADAMTS5; a disintegrin and metalloproteinase with thrombospondin motifs 5; MMP9, matrix metalloproteinase 9.

Article Snippet: Human MMP-9 Quantikine ELISA kit DMP900 (R&D Systems, Inc., Minneapolis, MN, USA) and Human ADAMTS5 DuoSet ELISA kit (R&D Systems, Inc.) were used according to the manufacture's protocols.

Techniques: Expressing, Clinical Proteomics

Journal: Burns & Trauma

Article Title: H3K56 lactylation promotes collagen II synthesis to modulate chondrocyte metabolism in posttraumatic osteoarthritis

doi: 10.1093/burnst/tkaf073

Figure Lengend Snippet: H3K56 lactylation in chondrocytes preserves cartilage matrix in mouse knee joints. ( a ) Immunohistochemistry of Col2a1, aggrecan, ADAMTS5, and MMP 13 in mouse knee joint (red arrow indicates the typically positive area, scale bar = 100 μm, n = 5). ( b) Relative positive area of Col2a1, aggrecan, ADAMTS5, and MMP 13 per section. Significance was determined by one-way ANOVA with Dunnett’s multiple comparison test (b). Data are mean ± SD from at least five experiments. ADAMTS5 a Disintegrin and Metalloproteinase with Thrombospondin motifs 5, MMP 13 matrix metalloproteinase 13, H3K56la histone H3 lysine 56 lactylation

Article Snippet: For western blotting analysis, antibodies against Col2a1 (AF6528, Beyotime, China), ADAMTS5 (bs-3573R, Bioss, China), Aggrecan (AF6126, Beyotime, China), MMP13 (AF7479, Beyotime, China), β-actin (AF0003, Beyotime, China), Histone H3 (AF0009, Beyotime, China), Histone H4 (AF2581, Beyotime, China), Pan Kla (PTM-1401, PTM, China), Pan Kac (PTM-101, PTM, China), H3K56la (PTM-1421RM, PTM, China), HIF 1α (#36169, CST, USA), HIF 1α (AF7087,Beyotime, China), HIF 1α (Huabio, China) were used.

Techniques: Immunohistochemistry, Comparison

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Flavokawain A alleviates the progression of mouse osteoarthritis: An in vitro and in vivo study

doi: 10.3389/fbioe.2022.1071776

Figure Lengend Snippet: Protective effects of FKA against IL-1β-induced inflammation and ECM destruction in mouse chondrocytes. Chondrocyte cell treatment with 5 ng/ml of IL-1β only or with FKA (20 and 40 μM) for a period of 24 h. Western blot analysis of inflammatory cytokines (COX2 and iNOS), catabolic (ADAMTS5, MMP3, and MMP13), and anabolic markers (Col2, Aggrecan, and Sox9) (A,C and E) . Quantitative analysis of protein expression (B,D and F) . (G and H) 5 ng/ml of IL-1β was added for treating the cells for 24 h in the absence or presence of 40 µM FKA. Confocal microscopy showing the expression of MMP13 and Aggrecan in immunofluorescence experiments (scale bar 10 μm). (I) Relative quantification of fluorescence intensity. The values are presented as means ± SD ( n = 3). # p < 0.05 vs the control group; * p < 0.05 and ** p < 0.01 vs the IL-1β group.

Article Snippet: ADAMTS5 primary antibody, secondary antibody, phosphate-buffered saline (PBS), trypsin, collagenase type II, the CCK8 assay kit, bovine serum albumin (BSA), and protein extraction kit were ordered and acquired from Boster Biological Technology (Wuhan, Hubei, China).

Techniques: Western Blot, Expressing, Confocal Microscopy, Immunofluorescence, Quantitative Proteomics, Fluorescence, Control