Journal: Bioactive Materials

Article Title: Bioengineered extracellular vesicles escape lysosomal degradation and deliver Tet-PKM2 for macrophage immunometabolic reprogramming and periodontitis treatment

doi: 10.1016/j.bioactmat.2026.01.002

Figure Lengend Snippet: Metabolic reprogramming and enhanced mitochondrial function in LPS-activated macrophages in response to LEV Tet−PKM2 @TA treatment. The macrophages were pretreated with 100 ng/mL LPS for 24 h and then treated with PBS (Control), 100 μg/mL LEVs PKM2 , LEVs Tet−PKM2 , or LEVs Tet−PKM2 @TA for another 24 h. ( A ) Heatmap representing differentially detected metabolites involved in glycolysis and the TCA cycle in the Control, LEVs PKM2 , LEVs Tet−PKM2 , or LEVs Tet−PKM2 @TA groups ( n = 4). ( B ) Concentrations of key glycolysis and TCA metabolites in Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( C ) Schematic illustration revealing changes in key glycolysis and TCA metabolites in the LEVs Tet−PKM2 @TA group versus the Control group. The up (down) arrows indicate increased (decreased) levels of metabolites in macrophages. ( D ) Kinetic profile of the ECAR in LPS-activated macrophages in response to sequential injections of glucose, oligomycin, and 2-DG in various groups (Seahorse XF test) ( n = 4). ( E ) Quantification of glycolysis, glycolytic capacity and glycolytic reserve in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( F ) Kinetic profile of the OCR in LPS-activated macrophages in response to sequential injections of oligomycin, FCCP, and Rot/AA in various groups (Seahorse XF test) ( n = 4). ( G ) Quantification of basal respiration, ATP production, and maximal respiration in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( H ) JC-1 aggregation (red fluorescence) in healthy mitochondria and cytosolic JC-1 monomers in compromised mitochondria (green fluorescence) (immunofluorescence assays). ( I ) Quantitative analysis of MMP levels determined by the relative ratio of red/green fluorescence intensity in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 4). ( J ) Intracellular ATP levels of LPS-activated macrophages in the Control, LEVs PKM2 , LEVs Tet−PKM2 , and LEVs Tet−PKM2 @TA groups ( n = 3). ( K-M ) The macrophages were pretreated with 100 ng/mL LPS for 24 h and then treated with PBS (Control), 10 μM UK-5099, 100 μg/mL LEVs Tet−PKM2 @TA, or 10 μM UK-5099 plus 100 μg/mL LEVs Tet−PKM2 @TA for another 24 h. ( K ) Schematic illustration revealing mechanism of LEVs Tet−PKM2 @TA promotes macrophage metabolic reprogramming depending on pyruvate influx into the TCA cycle. ( L ) Kinetic profile of the OCR in LPS-activated macrophages in response to sequential injections of oligomycin, FCCP, and Rot/AA in various groups (Seahorse XF test) ( n = 3). ( M ) Quantification of basal respiration, ATP production, and maximal respiration in the Control, UK-5099, LEVs Tet−PKM2 @TA, and UK-5099 + LEVs Tet−PKM2 @TA groups ( n = 3). The data are expressed as the mean ± SEM. Statistical analysis was performed with one-way ANOVA ( B , E , G, I, J , and M ). ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 indicate significant differences between the indicated columns.

Article Snippet: The MMP of the macrophages was assessed using a JC-1 MMP Assay Kit (MCE).

Techniques: Control, Fluorescence, Immunofluorescence

Journal: International Journal of Oncology

Article Title: Diverse roles of SERPINE1 in regulating cellular proliferation and invasion

doi: 10.3892/ijo.2026.5871

Figure Lengend Snippet: SERPINE1 regulates MMP activity. (A) MMP and TIMP levels in the supernatants of shSE1 and shc cells after 24 h of incubation and 10-fold concentration. Statistical significance was determined using a two-sided Student's t test. The data are presented as the means ± SDs. n=4. (B) Quantification of active MMP-1 and MMP-13 levels in cell lysates using fluorescence ELISA. Statistical significance was determined using two-way ANOVA followed by Šídák's multiple comparisons test. The data are presented as the means ± SDs of three independent experiments. (C) Western blotting showing the levels of the indicated proteins in H4-shSE1 cells at 72 h after transfection with the si-HSP90AA1 or siNC. The numbers beneath the bands represent the ratios of the expression levels of the indicated proteins and represent mean ± SDs of three independent experiments. (D) Representative images and quantification of Transwell invasion assays through Matrigel and collagen type I in H4-shSE1 cells transfected with the si-HSP90AA1 or siNC (scale bar, 100 μ m). Statistical significance was determined using a two-sided Student's t-test. The data are presented as the means ± SDs of three independent experiments. (E) Diagram showing the mechanisms underlying the effects of SERPINE1 on cancer proliferation and invasion. SERPINE1 knockdown influences cell proliferation and invasion through distinct signaling pathways. With respect to proliferation, SERPINE1 knockdown reduces TGF-β levels, and this reduction alters the activity of SMAD3, p53, and MCM3 to promote cell cycle progression. SERPINE1 knockdown interferes with the uPAR-mediated balance of the ERK/p38 ratio; it may also affect this ratio by modulating HSP90α expression and p38 activity, which suppress cell proliferation. In terms of invasion, SERPINE1 downregulation increases MMP-1 levels via the HSP90α-p38 pathway, thereby promoting cellular invasion. *** P<0.001, ** P<0.01, ns, not significant. SERPINE1, serine protease inhibitor clade e member 1; MMP, matrix metalloproteinases; TIMP, tissue inhibitors of metal proteases; shRNA, short hairpin RNA; shSE1, shRNA targeting SERPINE1; si, short interfering RNA; siHSP90AA1, short interfering heat shock protein 90 alpha family class a member 1; NC, negative control; uPAR, urokinase-type plasminogen activator receptor; p-, phosphorylated; MCM3, minichromosome maintenance complex component 3; HSP90α, heat shock protein 90-alpha.

Article Snippet: Following the blocking of nonspecific binding sites using 5% skimmed milk (cat. no. P0216; Beyotime Biotechnology) or bovine serum albumin (cat. no. NGP0028A; Beyotime Biotechnology) for 1 h at room temperature, the membranes were incubated with primary antibodies (incubation overnight at 4°C) against SERPINE1 (cat. no. 13801-1-AP; Proteintech Group, Inc), GAPDH (cat. no. sc-25778; Santa Cruz Biotechnology, Inc.), MCM3 (cat. no. PA5-79646; Thermo Fisher Scientific, Inc.), phosphorylated (p-)MCM3 (Ser112; cat. no. TA2362; Abmart Pharmaceutical Technology Co., Ltd.), ERK (cat. no. sc-514302; Santa Cruz Biotechnology, Inc.), uPAR (cat. no. ab10379; Abcam), Histone H3 (cat. no. 4499), p-p53 (Ser15; cat. no. 9284), p53 (cat. no. 2524), p-SMAD3 (Ser423/425; cat. no. 9520), SMAD3 (cat. no. 9523), p-Rb (Ser807/811; cat. no. 8516), Rb (cat. no. 9309), CyclinD1 (cat. no. 55506), CyclinE1 (cat. no. 20808), p21 (cat. no. 2947), p-p38 (cat. no. 4511), p38 (cat. no. 8690), p-ERK (cat. no. 4370), p-JNK (cat. no. 9255), JNK (cat. no. 9252), p-AKT (Ser473; cat. no. 4060), AKT (cat. no. 9272), HSP90α (cat. no. 4877) and MMP-1 (cat. no. 54376) were from Cell Signaling Technology, Inc.) and subsequently incubated with the corresponding secondary antibodies (dilution 1:4,000) for 2 h at room temperature.

Techniques: Activity Assay, Incubation, Concentration Assay, Fluorescence, Enzyme-linked Immunosorbent Assay, Western Blot, Transfection, Expressing, Knockdown, Protein-Protein interactions, Protease Inhibitor, shRNA, Small Interfering RNA, Negative Control

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: MMP-9 , a factor that promotes Vasculogenic mimicry, is highly expressed in CC and is associated with poor prognosis. (A) CC database of TCGA was used to analyze key factors associated with VM. (B) Association of Sox2 expression with overall survival in CC (log-rank test). (C) Association of MMP-9 expression with overall survival in CC (log-rank test). (D) Panoramic scans after immunohistochemical detection of MMP-9 and H&E staining in samples from cancerous and paracancerous tissues from subjects with CC. Scale bar, 50 µm. Original magnification, ×20. (E) Protein levels of MMP-9 in 20 paired samples, with the MMP-9 level in CC tissue expressed compared with that in the paired normal tissue. (F) Expression levels of MMP-9 mRNA in 44 paired CC and paracancerous tissues, with MMP-9 expression in CC tissue expressed compared with that in the paired normal tissue. (G) Comparison of the average expression levels of MMP-9 mRNA in CC tissues compared with paracancerous tissues. (H) HeLa and SiHa cells were incubated under hypoxia (0.1% O 2 ) and proteins collected at 24, 48 and 72 h for western blotting of ALDH1, EPHA2, MMP-9 and GAPDH. ImageJ was used to semi-quantify western blotting signals from HeLa (I) and SiHa (J) cells. GAPDH served as an internal reference. *P<0.05, **P<0.01 and ***P<0.001. MMP-9, matrix metalloproteinase 9; VM, vasculogenic mimicry; ALDH1, aldehyde dehydrogenase 1; EPHA2, ephrin type-A receptor 2; TCGA, The Cancer Genome Atlas; Sox2, SRY-box transcription factor 2; CC, cervical cancer; CESC, cervical squamous cell carcinoma.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Immunohistochemical staining, Staining, Comparison, Incubation, Western Blot

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 promotes Vasculogenic mimicry, invasion and migration of CC cells under hypoxic conditions. (A) Expression levels of NSUN2 in CC cells lines (HeLa, SiHa, CaSki, C33A and HT-3) and in a normal cervical cell line (HaCaT). (B) RT-qPCR was used to determine relative expression levels of NSUN2 mRNA in HeLa and SiHa cells after transfection of shRNAs and incubation under hypoxia for 24 h. (C) Relative expression levels of MMP-9 mRNA in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 24 h. (D) Dot blot assay analysis of m 5 C expression levels in HeLa and SiHa cells after 48 h of hypoxia culture following transfection with NSUN2 knockdown plasmids. (E) Semi-quantitative analysis of dot blot results in HeLa cells. (F) Semi-quantitative analysis of dot blot results in SiHa cells. (G) Western blotting was used to investigate NSUN2 and MMP-9 protein levels in HeLa and SiHa cells transfected with the NSUN2 -interfering plasmid and incubated under hypoxia for 48 h. ImageJ was used to semi-quantify western blotting bands for (H) NSUN2 and (I) MMP-9 protein levels. (J) 2D tube-forming assays of HeLa and SiHa cells transfected with an NSUN2 -interfering plasmid and incubated under hypoxia. Scale bar, 100 µm. Original magnification, ×4. (K) Assay displayed in panel (J) was quantified using Image Pro. (L) Invasion assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (M) Assay displayed in panel was quantified using Image Pro. (N) Migration assay of HeLa and SiHa cells transfected with a control plasmid, shNSUN2-2 or shNSUN2-3 or with shNSUN2 and pcDNA MMP-9. Scale bar, 200 µm. Original magnification, ×10. (O) Assay displayed in panel (N) was quantified using Image Pro. *P<0.05, **P<0.01 and ***P<0.001. VM, Vasculogenic mimicry; CC, cervical cancer; IHC, immunohistochemistry; NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; RT-qPCR, reverse transcription-quantitative PCR; IHC, immunohistochemistry; CaSki, human cervical cancer cell line with intestinal metastasis; C33A, human cervical cancer cell line; HaCaT, human skin keratinocytes cell line; HeLa, human cervical cancer cell line; HT-3, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Migration, Expressing, Quantitative RT-PCR, Transfection, Incubation, Plasmid Preparation, Dot Blot, Knockdown, Quantitative Dot Blot, Western Blot, Invasion Assay, Control, Immunohistochemistry, Reverse Transcription, Real-time Polymerase Chain Reaction, Cloning, shRNA, Negative Control

Journal: Oncology Letters

Article Title: RNA methyltransferase NSUN2 enhances vasculogenic mimicry and malignant progression of cervical cancer through upregulation of MMP-9

doi: 10.3892/ol.2026.15518

Figure Lengend Snippet: NSUN2 increases the stability of MMP-9 mRNA. (A) A positive correlation was observed between NSUN2 and MMP-9 mRNA expression levels in 44 pairs of samples from subjects with CC. Enrichment of the m 5 C modification of MMP-9 mRNA in HeLa (B) and SiHa (C) Cells were measured with anti-m 5 C methylated-RNA IP assays. Interaction of NSUN2 with MMP-9 mRNA in (D) HeLa and (E) SiHa cells was measured with anti- NSUN2 RNA IP assays. Stability of MMP-9 mRNA after interference with and overexpression of NSUN2 was measured in (F) HeLa and (G) SiHa cells. (H) A model illustrating the proposed mechanism by which NSUN2-mediated stabilization of MMP-9 mRNA promotes Vasculogenic mimicry in CC. NSUN2, NOP2/Sun RNA methyltransferase 2; m 5 C, 5-methylcytidine; pcDNA, plasmid cloning DNA; shRNA, short hairpin RNA; HeLa, human cervical cancer cell line; MMP9, matrix metalloproteinase 9; SiHa, human cervical squamous cell line; IP, immunoprecipitation; CC, cervical cancer; NC, negative control.

Article Snippet: The tissue samples were treated with PH9.0 EDTA repair solution for antigen retrieval and then treated with a rabbit polyclonal anti-CD31 antibody (1:2,000; cat. no. AB76533; Abcam), a rabbit polyclonal anti-NSUN2 antibody (1:200; cat. no. AB259941; Abcam) or a rabbit polyclonal anti-MMP-9 antibody (1:200; cat. no. 10375-2-AP; Proteintech Group, Inc.; Wuhan Sanying Biotechnology).

Techniques: Expressing, Modification, Methylation, Over Expression, Plasmid Preparation, Cloning, shRNA, Immunoprecipitation, Negative Control

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates the intestinal PPAR-γ and downregulates the MMP-9/2 expression. (A) The degree of the node between the HDCA and the intersection target. (B) Molecular docking analysis between HDCA and PPAR-γ. (C) Molecular docking analysis between HDCA and MMP-9. (D) Molecular docking analysis between HDCA and MMP-2. (E-G) Relative expression of mRNA of ppar-γ, mmp9 and mmp2 in the distal ileum. (n = 6) . (H) Representative protein immunoblots in distal ileum. (I-K) Relative expression of PPAR-γ, MMP-9, MMP-2 (n = 3–4). (L) Representative immunohistochemical staining and quantitative analysis of MMP-2+ (M), PPAR-γ+ (N) and MMP-9+ (O) cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (E–G, I–K, M–O). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates PPAR-γ expression and reduces MMP-9/2 expression in the spinal cord. (A–C) The relative expression of mRNA in spinal cord of ppar-γ, mmp9 and mmp2 . (n = 6) . (D–G) Representative immunoblots of proteins and relative expression of PPAR-γ, MMP-2, MMP-9 (n = 4). (H–J) Immunofluorescence staining of PPAR-γ, MMP-9, MMP-2 in spinal cord (scale, 100 μm). (n = 3). (K) Heatmap of Correlation Analysis. * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (A–D, F, G). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: Fxr knock down abolished the protective effect of HDCA in neuropathic pain. (A) Immunofluorescence staining showing FXR co-localization in spinal cord. (scale, 25 μm). (B) Immunofluorescence staining demonstrating MMP-2 colocalization in spinal cord. (scale, 25 μm). (C) Immunofluorescence staining showing PPAR-γ colocalization in spinal cord. (scale, 25 μm). (D) Immunofluorescence staining showing MMP-9 colocalization in spinal cord. (scale, 25 μm). (E) Immunofluorescence staining of FXR and MMP-2 colocalization in spinal cord. (scale, 25 μm). (F) Representative immunoblots of protein expression in the spinal cord from Fxr -/- mice. (G) Relative expression of PPAR-γ, MMP-2 and MMP-9 (n = 4). (H) Representative immunoblots for the proteins in spinal cord following INT-747 treatment. (I) PWT of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (J) PWL of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (K) Relative expression of PPAR-γ, MMP-2 and MMP-9 following INT-747 treatment. (n = 4). (L, M) Representative immunoblots for the proteins in spinal cord and relative expression of PPAR-γ, MMP-2 and MMP-9 in Fxr -/- mice following HDCA treatment. (n = 4). * P < 0.05, ** P < 0.01, ns by unpaired Student's t‐test (G, M). ns by Two-way repeated ANOVA with post hoc Bonferroni’s test (I and J), * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (K). Data are represented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Knockdown, Immunofluorescence, Staining, Western Blot, Expressing

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates the intestinal PPAR-γ and downregulates the MMP-9/2 expression. (A) The degree of the node between the HDCA and the intersection target. (B) Molecular docking analysis between HDCA and PPAR-γ. (C) Molecular docking analysis between HDCA and MMP-9. (D) Molecular docking analysis between HDCA and MMP-2. (E-G) Relative expression of mRNA of ppar-γ, mmp9 and mmp2 in the distal ileum. (n = 6) . (H) Representative protein immunoblots in distal ileum. (I-K) Relative expression of PPAR-γ, MMP-9, MMP-2 (n = 3–4). (L) Representative immunohistochemical staining and quantitative analysis of MMP-2+ (M), PPAR-γ+ (N) and MMP-9+ (O) cells in the distal ileum (scale bar, 100 μm, n = 6). * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (E–G, I–K, M–O). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunohistochemical staining, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: HDCA upregulates PPAR-γ expression and reduces MMP-9/2 expression in the spinal cord. (A–C) The relative expression of mRNA in spinal cord of ppar-γ, mmp9 and mmp2 . (n = 6) . (D–G) Representative immunoblots of proteins and relative expression of PPAR-γ, MMP-2, MMP-9 (n = 4). (H–J) Immunofluorescence staining of PPAR-γ, MMP-9, MMP-2 in spinal cord (scale, 100 μm). (n = 3). (K) Heatmap of Correlation Analysis. * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (A–D, F, G). Data is presented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Expressing, Western Blot, Immunofluorescence, Staining

Journal: Journal of Advanced Research

Article Title: Hyodeoxycholic acid relieves neuropathic pain by activating farnesoid X receptor signaling

doi: 10.1016/j.jare.2025.07.017

Figure Lengend Snippet: Fxr knock down abolished the protective effect of HDCA in neuropathic pain. (A) Immunofluorescence staining showing FXR co-localization in spinal cord. (scale, 25 μm). (B) Immunofluorescence staining demonstrating MMP-2 colocalization in spinal cord. (scale, 25 μm). (C) Immunofluorescence staining showing PPAR-γ colocalization in spinal cord. (scale, 25 μm). (D) Immunofluorescence staining showing MMP-9 colocalization in spinal cord. (scale, 25 μm). (E) Immunofluorescence staining of FXR and MMP-2 colocalization in spinal cord. (scale, 25 μm). (F) Representative immunoblots of protein expression in the spinal cord from Fxr -/- mice. (G) Relative expression of PPAR-γ, MMP-2 and MMP-9 (n = 4). (H) Representative immunoblots for the proteins in spinal cord following INT-747 treatment. (I) PWT of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (J) PWL of Fxr -/- mice following HDCA or INT-747 treatment. (n = 6). (K) Relative expression of PPAR-γ, MMP-2 and MMP-9 following INT-747 treatment. (n = 4). (L, M) Representative immunoblots for the proteins in spinal cord and relative expression of PPAR-γ, MMP-2 and MMP-9 in Fxr -/- mice following HDCA treatment. (n = 4). * P < 0.05, ** P < 0.01, ns by unpaired Student's t‐test (G, M). ns by Two-way repeated ANOVA with post hoc Bonferroni’s test (I and J), * P < 0.05, ** P < 0.01, *** P < 0.001 by One-way ANOVA with post hoc Tukey's test (K). Data are represented as mean ± SEM.

Article Snippet: Tissue sections of each group were blocked with 1 % bovine serum albumin and 10 % donkey serum at room temperature for 1 h and then incubated at 4 °C overnight with primary antibodies for neuronal nuclear protein (NeuN) (1:500, Abcam, ab104224), ionized calcium-binding adapter molecule 1 (IBA-1) (1:500, Abcam, ab5076), glial fibrillary acidic protein (GFAP) (1:500, Millipore, MAB360), FXR (1:200, Proteintech, 25055–1-AP), MMP-2 (1:200, Proteintech, 10373-2-AP), MMP-9 (1:200, Proteintech, 10375-2-AP), PPAR-γ (1:200, Proteintech, 16643–1-AP), NLRP3 (1:200, Immunoway, YT5382), and cluster of differentiation 86 (CD86) (1:200, CST, #91882).

Techniques: Knockdown, Immunofluorescence, Staining, Western Blot, Expressing