Journal: Biomedical reports

Article Title: Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells.

doi: 10.3892/br.2023.1681

Figure Lengend Snippet: Figure 4. Effect of Oct‑LMSG on type I collagen mRNA transcription in L929 fibroblasts. The mRNA levels of Col1A1 and Col1A2 were determined by reverse transcription‑quantitative PCR assay after treatment with either Oct‑LMSG (100 µg/ml) or TGF‑β (10 ng/ml) for 6, 12 and 24 h relative to GAPDH. Values are expressed as the mean ± standard error of the mean of three independent experiments. *P<0.05 vs. control. Oct‑LMSG, low molecular weight sulfated galactan added with octanoyl moiety; Col1A1, collagen type I α1 chain.

Article Snippet: After 12 h of treatment, cells were incubated with anti‐collagen type I α1 chain (Col1A1) antibody (dilution, 1:250; cat. no. A22089; ABclonal Science, Inc.) overnight at 4 ̊C, followed by incu‐ bation with the secondary antibody, FITC‐conjugated goat anti‐rabbit IgG (dilution, 1:500; cat. no. 701078; Thermo Fisher Scientific, Inc.), for 1 h at RT.

Techniques: Control, Molecular Weight

Journal: Biomedical reports

Article Title: Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells.

doi: 10.3892/br.2023.1681

Figure Lengend Snippet: Figure 5. Effect of Oct‑LMSG on type I collagen protein expression in L929 fibroblasts. The Col1A1 and Col1A2 protein levels were detected by western blot assay after treatment with either Oct‑LMSG (100 µg/ml) or TGF‑β (10 ng/ml) for 6, 12 and 24 h relative to β‑actin. Values are expressed as the mean ± standard error of the mean of three independent experiments. *P<0.05 vs. control. Oct‑LMSG, low molecular weight sulfated galactan added with octanoyl moiety; Col1A1, collagen type I α1 chain.

Article Snippet: After 12 h of treatment, cells were incubated with anti‐collagen type I α1 chain (Col1A1) antibody (dilution, 1:250; cat. no. A22089; ABclonal Science, Inc.) overnight at 4 ̊C, followed by incu‐ bation with the secondary antibody, FITC‐conjugated goat anti‐rabbit IgG (dilution, 1:500; cat. no. 701078; Thermo Fisher Scientific, Inc.), for 1 h at RT.

Techniques: Expressing, Western Blot, Control, Molecular Weight

Journal: Biomedical reports

Article Title: Octanoyl esterification of low molecular weight sulfated galactan enhances the cellular uptake and collagen expression in fibroblast cells.

doi: 10.3892/br.2023.1681

Figure Lengend Snippet: Figure 6. Effect of Oct‑LMSG on collagen production in L929 fibroblasts. The collagen synthesis was observed by Col1A1‑immunofluorescence staining assay. Fluorescent micrographs of Col1A1‑stained fibroblasts. Cells were treated with either Oct‑LMSG (100 µg/ml) or TGF‑β (10 ng/ml) for 12 h and stained with Col1A1, followed by secondary conjugation with FITC antibodies. Cells were counterstained with DAPI (nucleus) and CellMask Deep Red (cell cytoskeleton). Collagen‑Col1A1 is represented by fluorescent green, DAPI by fluorescent blue and CellMask Deep Red by fluorescent red. The negative control was prepared by omission of the secondary antibody, resulting in the absence of fluorescent green (data not shown). Merged fluorescent micrographs are also provided (scale bars, 75 µm). Oct‑LMSG, low molecular weight sulfated galactan added with octanoyl moiety; Col1A1, collagen type I α1 chain.

Article Snippet: After 12 h of treatment, cells were incubated with anti‐collagen type I α1 chain (Col1A1) antibody (dilution, 1:250; cat. no. A22089; ABclonal Science, Inc.) overnight at 4 ̊C, followed by incu‐ bation with the secondary antibody, FITC‐conjugated goat anti‐rabbit IgG (dilution, 1:500; cat. no. 701078; Thermo Fisher Scientific, Inc.), for 1 h at RT.

Techniques: Staining, Conjugation Assay, Negative Control, Molecular Weight

Journal: International Journal of Molecular Medicine

Article Title: Orcinol glucoside ameliorates pulmonary fibrosis by suppressing hyaluronic acid synthesis and macrophage M2 polarization via targeting hyaluronic acid synthase 2

doi: 10.3892/ijmm.2026.5764

Figure Lengend Snippet: Elevated HAS2 expression and HA accumulation in murine and cellular fibrosis models. (A) Schematic of the experimental design for the time-course study. C57BL/6J mice received a single intratracheal dose of BLM or saline and were sacrificed at the indicated time points (days 0, 3, 5, 7, 10, 14 and 21) for sample collection (n=5 for per group). (B) Representative western blot images (left panel) and densitometric quantification (right panel) showing the protein expression levels of COL1A1 and HAS2 in lung tissues across the time course. β-tubulin served as the loading control (n=4 for per group). (C) ELISA quantification of HA levels in BALF at different days post-BLM injury (n=5 for per group). (D) Schematic diagram illustrating the workflow for the isolation and culture of primary mouse lung fibroblasts from BLM-treated fibrotic mice. (E) Western blotting analysis of HAS2 and COL1A1 protein expression in primary lung fibroblasts isolated from control (saline) and BLM-induced fibrotic mice. Representative blot images (left panel) and densitometric quantification of protein levels normalized to β-tubulin are shown (n=3 for each group). (F) Schematic diagram depicting TGF-β1-induced transition of NIH/3T3 fibroblasts into myofibroblasts. (G) COL1A1, ACTA2, and HAS2 mRNA expression in NIH/3T3 cells was detected using PCR after 5 ng/ml TGF-β1 administration for 12 h (n=3-4 for each group). (H) HAS2 protein expression in NIH/3T3 was detected using western blotting after 5 ng/ml TGF-β1 administration for 24 h (n=3 for each group). (I) ELISA was used to quantify HA concentrations in the culture media of myofibroblasts (n=6 for each group). * P<0.05, ** P<0.01, *** P<0.001. HAS2, hyaluronic acid synthase 2; HA, hyaluronic acid; BLM, bleomycin; BALF, bronchoalveolar lavage fluid; COL1A1, Collagen type I α 1 chain; ACTA2, actin alpha 2, smooth muscle.

Article Snippet: Antibodies for Collagen type I α 1 chain (COL1A1; cat. no. 72026; 1:1,000), Phospho-STAT6 (Tyr641; cat. no. 56554S; 1:1,000) and α-smooth muscle actin (α-SMA; cat. no. 19245; 1:1,000) were obtained from Cell Signaling Technology, Inc.

Techniques: Expressing, Saline, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Isolation

Journal: International Journal of Molecular Medicine

Article Title: Orcinol glucoside ameliorates pulmonary fibrosis by suppressing hyaluronic acid synthesis and macrophage M2 polarization via targeting hyaluronic acid synthase 2

doi: 10.3892/ijmm.2026.5764

Figure Lengend Snippet: OG exerts an anti-pulmonary fibrosis effect by impairing myofibroblast-mediated macrophage M2 polarization. (A) Schematic workflow: Fibroblast treatment with OG during TGF-β1 stimulation, followed by conditioned medium collection and macrophage culture for downstream assays. (B) TGF-β1 release from macrophages measured by ELISA (n=3 for each group). (C) Flow cytometric analysis of CD206 and CD86 expression in macrophages cultured in conditioned medium. The bar graph quantifies the proportion of CD206 + CD86 − cells (M2-like macrophage subpopulation; n=3 for each group). (D) Animal experimental protocol. (E) A BLM-induced mouse model was established to assess the anti-fibrotic effects of OG. Histopathological alterations in lung tissues from different groups were assessed using H&E and Masson's trichrome staining. Scale bars: 500 μ m; magnification, ×40. (F) Western blotting analysis to measure the expression levels of fibrotic markers, COL1A1 and α-SMA, in lung tissues (n=4 for each group). (G) The levels of HA in mouse serum were quantified by ELISA. (H) The levels of TGF-β1 in mouse serum were quantified by ELISA (n=6 for each group). (I) OG concentrations in serum and lung tissues after 14-day oral administration. Left: serum concentrations of OG across treatment groups. Right: lung tissue concentrations of OG, expressed as a percentage of tissue weight (n=3 for each group). * P<0.05, ** P<0.01, *** P<0.001. OG, glucoside; BLM, bleomycin; COL1A1, Collagen type I α 1 chain; α-SMA, α-smooth muscle actin; H&E, hematoxylin and eosin.

Article Snippet: Antibodies for Collagen type I α 1 chain (COL1A1; cat. no. 72026; 1:1,000), Phospho-STAT6 (Tyr641; cat. no. 56554S; 1:1,000) and α-smooth muscle actin (α-SMA; cat. no. 19245; 1:1,000) were obtained from Cell Signaling Technology, Inc.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Cell Culture, Staining, Western Blot

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Interleukin 10 Attenuates Angiotensin II-Induced Aortic Remodelling by Inhibiting Oxidative Stress-Induced Activation of the Vascular p38 and NF- κ B Pathways

doi: 10.1155/2022/8244497

Figure Lengend Snippet: IL-10 suppresses Ang II-induced proliferation, fibrosis, and inflammation of mouse vascular adventitial fibroblasts (mVAFs). (a) and (b) Representative images of immunofluorescence colocalization staining of Ki67 (green) and DAPI (navy) in mVAFs treated with Ang II ( n =8 for each group; scale bars, 100 μ m). (c)–(j) The mRNA levels of inflammatory markers (TNF- α , IL-1 β , IL-6, and IL-8) and fibrotic markers (Col1 α 1, Col3 α 1, TGF- β , and CTGF) in mVAFs treated with Ang II were assessed by real-time PCR. (k)–(p) The protein expression levels of PCNA, TNF- α , and fibrotic markers were examined by western blotting in mVAFs treated with Ang II. Relative protein levels were normalized to GAPDH. The data are representative of three independent experiments. Error bars show S.E. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001.

Article Snippet: Collagen type I α 1 (Colla1, cat. no. ab34710), TGF- β (cat. no. ab179695), TNF alpha (TNF- α , cat. no. ab6671), alpha smooth muscle actin ( α -SMA, cat. no. ab32575), and Ki67 (cat. no. ab16667) were purchased from Abcam (Cambridge, MA, USA).

Techniques: Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Expressing, Western Blot