Journal: Pharmaceutics

Article Title: Colon-Targeted Poly(ADP-ribose) Polymerase Inhibitors Synergize Therapeutic Effects of Mesalazine Against Rat Colitis Induced by 2,4-Dinitrobenzenesulfonic Acid.

doi: 10.3390/pharmaceutics16121546

Figure Lengend Snippet: Figure 3. AQSA-Glu potentiates the anticolitic activity of 5-AIQ. Three days after colitis induction by DNBS, 5-AIQ (5 mg/kg) and AQSA-Glu [7.5 mg/kg, equivalent to 2.5 mg/kg of 5-AIQ (L) and 15 mg/kg, equivalent to 5 mg/kg of 5-AIQ (H)] were administered orally to rats once per day, and the rats were euthanized 24 h after the sixth treatment. (A) Left panel: photos of the distal colons of rats in which serosal and luminal sides are shown separately. Right panel: overall colonic damage was scored for each group and presented as colonic damage score (CDS). * α < 0.05 vs. DNBS control. (B) H & E staining was performed with the colonic tissue sections of rats subjected to various treatments. Upper panel: representative images of 100× magnification. Lower panel: representative images of 200× magnification for the dotted boxes in the upper panel. In the inflamed distal colons (4.0 cm), (C) myeloperoxidase (MPO) activity was measured in addition to determining the levels of (D) CINC-3 and (E) iNOS and COX-2 using an Elisa kit and Western blotting. A loading control (α-Tubulin) was used for Western blot analysis of COX-2 and iNOS. NM: not measurable. The data are represented as mean ± SD (n = 5). * p < 0.05 vs. DNBS control # p < 0.05.

Article Snippet: Nrf2, HO-1, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) proteins were detected using the following antibodies: anti-Nrf2 (sc-365949, Santa Cruz Biotechnology, Dallas, TX, USA), anti-HO-1 (sc-136961, Santa Cruz Biotechnology), anti-COX-2 (sc-365374, Santa Cruz Biotechnology), anti-iNOS (NOS-2) antibody (sc-7271, Santa Cruz Biotechnology), and secondary antibodies corresponding to the primary antibodies (Santa Cruz Biotechnology).

Techniques: Activity Assay, Control, Staining, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Pharmaceutics

Article Title: Colon-Targeted Poly(ADP-ribose) Polymerase Inhibitors Synergize Therapeutic Effects of Mesalazine Against Rat Colitis Induced by 2,4-Dinitrobenzenesulfonic Acid.

doi: 10.3390/pharmaceutics16121546

Figure Lengend Snippet: Figure 4. Colon-targeted PARP inhibitors synergize the anticolitic effects of mesalazine. (A) RAW264.7 cells pretreated with 5-ASA (20 mM), 3-AB (1 mM), and 5-AIQ (10 µM) for 1 h were challenged with LPS for 24 h. The levels of iNOS and COX-2 proteins were analyzed using West- ern blotting. (B) SSZ (50 mg/kg) and ABSA (36 mg/kg, equimolar to 50 mg/kg of SSZ) suspended in PBS (1 mL) were administered orally to rats. The rats were killed 2, 4, and 8 h after oral administration. The concentrations of 5-ASA in the cecum were analyzed using HPLC. (C) Three days after colitis induction by DNBS, SSZ (50 mg/kg), AQSA-Glu (15 mg/kg), a mixture of AQSA-Glu (15 mg/kg) + olsalazine (OSZ, 19 mg/kg, half-equimolar to 50 mg/kg of SSZ), and ABSA (36 mg/kg, equimolar to 50 mg/kg of SSZ) were administered orally to rats once per day, and the rats were euthanized 24 h after the sixth treatment. (C) Left panel: photos of the distal colons of rats where serosal and luminal sides are shown separately. Right panel: overall colonic damage was scored for each group and presented as colonic damage score (CDS). * α < 0.05 vs. DNBS control. (D) H & E staining was performed with the colonic tissue sections of rats subjected to various treatments. Upper panel: rep- resentative images of 100× magnification. Lower panel: representative images of 200× magnification for the dotted boxes in the upper panel. In the inflamed distal colons (4.0 cm), (E) myeloperoxidase (MPO) activity was measured in addition to determining the levels of (F) CINC-3 and (G) iNOS and COX-2 using an Elisa kit and Western blotting. A loading control (α-Tubulin) was used for Western blot analysis of COX-2 and iNOS. NM: not measurable. The data are represented as mean ± SD (n = 5). * p < 0.05 vs. DNBS control # p < 0.05.

Article Snippet: Nrf2, HO-1, cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS) proteins were detected using the following antibodies: anti-Nrf2 (sc-365949, Santa Cruz Biotechnology, Dallas, TX, USA), anti-HO-1 (sc-136961, Santa Cruz Biotechnology), anti-COX-2 (sc-365374, Santa Cruz Biotechnology), anti-iNOS (NOS-2) antibody (sc-7271, Santa Cruz Biotechnology), and secondary antibodies corresponding to the primary antibodies (Santa Cruz Biotechnology).

Techniques: Control, Staining, Activity Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Pharmacological research

Article Title: CircRNA CDR1as affects functional repair after spinal cord injury and regulates fibrosis through the SMAD pathway.

doi: 10.1016/j.phrs.2024.107189

Figure Lengend Snippet: Fig. 4. CDR1as regulates NSCs differentiation and microglia polarization. NSCs differentiation was identified by immunofluorescence. GFAP staining was used to detect astrocytes and MAP2 was used to detect nerve cells (A-B). The differentiation was further identified by PCR. The characteristics of nerve cells included MAP2, Nestin and TUJ1, and the characteristics of astrocytes were GFAP (C). iNOS (D) and Arg-1 (E) were detected by immunofluorescence after the BV2 model was established in vitro. Quantitative analysis of immunofluorescence staining (F). Expression of proinflammatory cytokines iNOS, IL-6, TNF-α (G) and anti-inflammatory cytokines Arg-1, CD206 and IL-4 (H) by PCR. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: The immunofluorescence steps of NSCs and BV2 differentiation are consistent with the identification of fibroblast, and primary antibodies were: inducible nitric oxide synthase (iNOS,6022, Proteintech, Rosemont, USA), Arginase-1 (Arg-1, 16001–1-AP, Proteintech), GFAP (3670, CST), and microtubule-associated protein 2 (MAP2, 17490–1-AP, Proteintech).

Techniques: Immunofluorescence, Staining, In Vitro, Expressing

Journal: International immunopharmacology

Article Title: KLF15 regulates macrophage polarization patterns in deep vein thrombosis.

doi: 10.1016/j.intimp.2025.114632

Figure Lengend Snippet: Fig. 1. Macrophage polarization patterns and KLF15 expression in DVT. A-B: ELISA kits were used to measure plasma levels of P-selectin (A) and vWF (B) in Sham and DVT group mice (N = 3). C-D: western blotting (C) and IF staining (D) were used to examine the expression and localization of M1-like macrophage (iNOS) and M2-like macrophage (CD206) markers in the inferior vena cava (IVC) tissues of mice in the Sham and DVT groups (N = 3). Scale = 100 μm. E: serum levels of pro-inflammatory factors (IL-1β and IL-6) and anti-inflammatory factors (IL-10 and TGF-β) released by M1/M2-like macrophages in mice from Sham and DVT groups were detected using ELISA kits (N = 3). F: Representative images of KLF15 IHC staining of IVC tissues from Sham and DVT group mice (N = 3). Scale = 500 μm. G-H: Western blotting (G) and RT-qPCR (H) assays were performed to detect the levels of KLF15 protein and mRNA in IVC tissues (N = 3).

Article Snippet: PVDF membranes were incubated in hybridization bags with antibodies, including rabbit KLF15 pAb, rabbit CD206 pAb (18704-1- AP; Proteintech; 1:5000), mouse iNOS mAb, rabbit NEK2 pAb (24171-1- AP; Proteintech; 1:1000), rabbit recombinant P65 (80979-1-RR; Proteintech; 1:40,000), mouse β-actin mAb (TA-09; ZSGB, China; 1:4000), rabbit p –P65 pAb (bs-5661R; Bioss, China; 1:2000), goat anti-mouse/ rabbit IgG-HRP (M21001L/M21002L; Abmart; 1:4000).

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Western Blot, Staining, Immunohistochemistry, Quantitative RT-PCR

Journal: International immunopharmacology

Article Title: KLF15 regulates macrophage polarization patterns in deep vein thrombosis.

doi: 10.1016/j.intimp.2025.114632

Figure Lengend Snippet: Fig. 2. KLF15 overexpression promotes M1 polarization in macrophages. A-B: the expression differences of KLF15 protein and mRNA in macrophages of PMA, M1, M1-NC and M1-KLF15 groups were detected using western blotting (A) and RT-qPCR (B) assays (N = 3). C: Effect of KLF15 overexpression on the proportion of CD86+ macrophages detected by flow cytometry (N = 3). D-E: Effect of KLF15 overexpression on the expression of M1 macrophage marker (iNOS) protein (D) and mRNA (E) (N = 3). F: representative images of iNOS IF staining in macrophages (N = 3). Scale = 100 μm. G: levels of pro-inflammatory factors (IL-1β and IL-6) in the supernatants of macrophages from the PMA, M1, M1-NC and M1-KLF15 groups were detected by ELISA kits (N = 3). H: Effect of KLF15 overexpression on IL-12 and TNF-α levels in M1 macrophages (N = 3).

Article Snippet: PVDF membranes were incubated in hybridization bags with antibodies, including rabbit KLF15 pAb, rabbit CD206 pAb (18704-1- AP; Proteintech; 1:5000), mouse iNOS mAb, rabbit NEK2 pAb (24171-1- AP; Proteintech; 1:1000), rabbit recombinant P65 (80979-1-RR; Proteintech; 1:40,000), mouse β-actin mAb (TA-09; ZSGB, China; 1:4000), rabbit p –P65 pAb (bs-5661R; Bioss, China; 1:2000), goat anti-mouse/ rabbit IgG-HRP (M21001L/M21002L; Abmart; 1:4000).

Techniques: Over Expression, Expressing, Western Blot, Quantitative RT-PCR, Flow Cytometry, Marker, Staining, Enzyme-linked Immunosorbent Assay