Journal: Scientific Reports

Article Title: Change in the chondroitin/dermatan structure in distal lung tissue from COPD patients

doi: 10.1038/s41598-026-44120-4

Figure Lengend Snippet: Effect of TGF-β activity. ( A ) Gene set enrichment analysis (GSEA) of ranked list generated from differential expression of RNAseq data from cohort 2 was performed on COPD patients (GOLD stage I-II, n = 96) versus controls (n = 91) lung tissue enriched against cellular response to TGF-β stimulus. ( B ) Pearson and Spearman correlation between CHST11 and TGF-β score generated using enrichment scoring for each sample. Correlation of all GAG enzymes are found in Supplemental Fig. . ( C ) Quantification of proteoglycans in HFL1 cells following TGF-β stimulation measured using SRM technology with pre-chosen peptides described in supplemental Table , (n = 4). ( D ) Quantification of GAGs in HFL1 cell medium following TGF-β stimulation (n = 4). ( E ) Quantification of HS disaccharides in HFL1 cell medium following TGF-β stimulation (n = 4). ( F ) Quantification of CS/DS disaccharides in HFL1 cell medium following TGF-β stimulation, (n = 4). ( G ) Quantification of DS disaccharides in HFL1 cells following TGF-β stimulation using AMAC-labelling after chondroitinase B digestion (n = 4). ( H ) mRNA fold change of selected CS/DS biosynthetic enzymes in TGF-β stimulated HFL1 cells compared to vehicle control measured by qPCR, (n = 4). Data is presented as mean ± S.D., * p < 0.05, ** p < 0.01 and *** P < 0.001 using Student’s T-test.

Article Snippet: HFL1 cells were obtained from ATCC (American Type Culture Collection).

Techniques: Activity Assay, Generated, Quantitative Proteomics, RNA sequencing, Control

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) prevent deterioration of renal function in vivo. a Representative images of renal histology (200×, scale bar = 50 μm). b The histomorphological score. c Immunohistochemical analysis of PCNA expression in kidney tissues (200×, scale bar = 50 μm). d Percentage of proliferating cell nuclear antigen ( PCNA )-positive cells. e TUNEL assay for apoptotic cells in rat renal tissues (200×, scale bar = 50 μm). f Percentage of apoptotic cells. Serum creatinine ( Cr ) ( g ) and blood urea nitrogen ( BUN ) ( h ) levels were measured at different times after cisplatin injection. Data are expressed as mean ± SD. * P < 0.05, n = 6. Rats treated as mentioned for the in vivo rat model were divided into four groups as follows: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; and 4) human fetal lung fibroblast-1exosomes ( HFL1-Ex ). All animals were sacrificed at 3 days after cisplatin injection

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing, TUNEL Assay, Injection, Control, Saline

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) inhibit cisplatin-induced apoptosis and secretion of inflammatory cytokines in NRK-52E cells in vitro. a NRK-52E cells were stained with JC-1 dye and observed under fluorescent microscopy (JC-1 produces red fluorescence within the mitochondria as JC-1-aggregates while it emits green fluorescence when it leaks into the cytoplasm as JC-1-monomers; the shift between green and red is proportional to the mitochondrial membrane potential change) (200×, scale bar = 50 μm). b TUNEL assay was performed to detect the apoptotic cells (200×, scale bar = 50 μm). c Percentage of apoptotic cells. ** P < 0.01, n = 3. d The expression of proliferating cell nuclear antigen ( PCNA ), BCL-XL, BCL-2, Bax, Cytochrome-c, cleaved Caspase-3, and GAPDH proteins was evaluated by Western blot. e Immunohistochemical analysis of tumor necrosis factor alpha ( TNF-α ) expression (200×, scale bar = 50 μm). f The expression of interleukin 1-β ( IL1-β ) and GAPDH proteins was evaluated by Western blot. NRK-52E cells treated as mentioned for the in vitro experiment were divided into four groups: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; and 4) human fetal lung fibroblast-1exosomes ( HFL1-Ex )

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vitro, Staining, Microscopy, Fluorescence, Membrane, TUNEL Assay, Expressing, Western Blot, Immunohistochemical staining, Control, Saline

Journal: Stem Cell Research & Therapy

Article Title: Pre-incubation with hucMSC-exosomes prevents cisplatin-induced nephrotoxicity by activating autophagy

doi: 10.1186/s13287-016-0463-4

Figure Lengend Snippet: Human umbilical cord-derived mesenchymal stem cell exosomes ( hucMSC-Ex ) prevent apoptosis and secretion of inflammatory cytokines by activating autophagy in vivo. a Representative images of renal histology (200×, scale bar = 50 μm). b The histomorphological score. c Immunohistochemical analysis of PCNA expression in kidney tissues (200×, scale bar = 50 μm). d Percentage of proliferating cell nuclear antigen ( PCNA )-positive cells. e TUNEL assay for apoptotic cells in rat renal tissues (200×, scale bar = 50 μm). f Percentage of apoptotic cells. g The expression of LC3B, BCL-2, Bax, IL1-β, and GAPDH proteins in the cisplatin-induced rat model were measured by Western blot. The serum inflammatory cytokines ( h ) tumor necrosis factor alpha ( TNF-α ), ( i ) interleukin-1β ( IL1-β ), and ( j ) interleukin-6 ( IL6 ) were measured via Luminex assay. Data are expressed as mean ± SD. * P < 0.05, n = 6. Rats were divided into six groups as follows: 1) control; 2) phosphate-buffered saline ( PBS ); 3) hucMSC-Ex; 4) hucMSC-Ex + 3-methyladenine ( 3MA ); 5) 3MA; and 6) rapamycin ( Rapa )

Article Snippet: Terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) staining was used to detect the apoptotic cells. hucMSCs, HFL1 cells, NRK-52E cells, and the kidney tissues were examined using the TdT-FragEL DNA fragmentation detection kit according to the manufacturer's protocol (Boster, China).

Techniques: Derivative Assay, In Vivo, Immunohistochemical staining, Expressing, TUNEL Assay, Western Blot, Luminex, Control, Saline

Journal: Scientific Reports

Article Title: Plant miRNA osa-miR172d-5p suppressed lung fibrosis by targeting Tab1

doi: 10.1038/s41598-023-29188-6

Figure Lengend Snippet: Osa-miR172d-5p identified as a plant miR candidate with an anti-fibrotic effect. ( A ) Scheme of plant miR selection. ( B ) In silico analysis of the interaction between osa-miR172d-5p and TAB1. ( C ) Human lung fibroblast HFL1 cells were transfected with the osa-miR172d-5p for 48 h, and TAB1 expression was evaluated via western blot analysis ( n = 4). ( D ) HFL1 cells were transfected with the indicated concentration of osa-miR172d-5p for 48 h, and cDNA was evaluated via qRT-PCR ( n = 4). Data are shown as mean ± SEM. * P < 0.05. *** P < 0.001 versus control group.

Article Snippet: Human lung fibroblast HFL1 cells (JCRB, Osaka, Japan) were cultured in 10% fetal bovine serum (FBS) (Sigma-Aldrich), Dulbecco's Modified Eagle Medium (DMEM) (044-29765; Fujifilm Tokyo, Japan) supplemented with penicillin G (876111)–streptomycin (876161; Meiji Pharmaceutical Co., Tokyo, Japan) in 5% CO 2 and 100% humidity at 37 °C.

Techniques: Selection, In Silico, Transfection, Expressing, Western Blot, Concentration Assay, Quantitative RT-PCR, Control

Journal: Scientific Reports

Article Title: Plant miRNA osa-miR172d-5p suppressed lung fibrosis by targeting Tab1

doi: 10.1038/s41598-023-29188-6

Figure Lengend Snippet: TAB1 knockdown suppressed TGFβ-induced fibrotic gene expression. ( A ) Human lung fibroblast HFL1 cells were transfected with TAB1-siRNA (10 nM, 48 h) and TAB1 expression levels were determined by western blot analysis. ( B – D ) Human lung fibroblast HFL1 cells were transfected with TAB1-siRNA (10 nM, 48 h) and treated with TGFβ (5 ng/mL for B , D 48 h; C 24 h). mRNA expression levels were assessed via RT-qPCR. ( B ) ASMA (αSMA) (n = 4), ( C ) COL1A1 (n = 4), and ( D ) FN (fibronectin; n = 4). Data are shown as mean ± SEM. * P < 0.05. ** P < 0.01. *** P < 0.001 versus control group.

Article Snippet: Human lung fibroblast HFL1 cells (JCRB, Osaka, Japan) were cultured in 10% fetal bovine serum (FBS) (Sigma-Aldrich), Dulbecco's Modified Eagle Medium (DMEM) (044-29765; Fujifilm Tokyo, Japan) supplemented with penicillin G (876111)–streptomycin (876161; Meiji Pharmaceutical Co., Tokyo, Japan) in 5% CO 2 and 100% humidity at 37 °C.

Techniques: Knockdown, Gene Expression, Transfection, Expressing, Western Blot, Quantitative RT-PCR, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: Macrophage‐derived exosomes modulate myofibroblast differentiation in vitro. (A) Exosomes derived from RAW264.7 cells were labelled with PKH26 dye and then incubated with HFL1 cells for 24 h. Scale bar: 20 μm. (B, D‐E) Western blot (B) and RT‐qPCR (D‐E) analyses of collagen I, α‐SMA and GAPDH in HFL1 cells treated with PBS, NC‐Exos, SiO 2 ‐Exos or SiO 2 + GW4869‐Exos were performed. These exosomes were derived from THP‐1 cells. (C, F‐G) Western blot (C) and RT‐qPCR (F‐G) analyses of collagen I, α‐SMA and GAPDH in NIH‐3T3 cells incubated with PBS, NC‐Exos, SiO 2 ‐Exos or SiO 2 + GW4869‐Exos were performed. These exosomes were derived from RAW264.7 cells. Student's t test; * P < .05, ** P < .01

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Derivative Assay, In Vitro, Incubation, Western Blot, Quantitative RT-PCR

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: SiO 2 ‐Exos induce lung fibroblast proliferation and migration (A‐B) CCK‐8 assay was used to evaluate the viability of HFL1 cells (A) and NIH‐3T3 cells (B) treated with various exosomes (SiO 2 ‐Exos or SiO 2 + GW4869‐Exos). Two‐way ANOVA; ** P < .01, *** P < .001, n.s: not significant. (C‐D) The migration of HFL1 cells (C) and NIH‐3T3 cells (D) was assessed by wound closure assay. Scale bar: 100 μm. Student's t test; * P < .05, ** P < .01, *** P < .001, n.s: not significant

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Migration, CCK-8 Assay, Wound Closure Assay

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: SiO 2 ‐Exo‐induced myofibroblast differentiation is ER stress dependent (A‐C, E) Western blot analysis of BIP, XBP1s, P ‐eIF2α, α‐SMA, Collagen I, and GAPDH in HFL1 cells (A, C) and NIH‐3T3 cells (B, E) incubated with SiO 2 ‐Exos (± 4‐PBA) at different points in time (0, 8, 16 or 24 h). (D, F) Analysis of collagen I, α‐SMA, and GAPDH in HFL1 cells (D) and NIH‐3T3 cells (F) incubated with SiO 2 ‐Exos or SiO 2 ‐Exos +4‐PBA for 24 h

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Western Blot, Incubation

Journal: Journal of Cellular and Molecular Medicine

Article Title: Macrophage‐derived exosomes mediate silica‐induced pulmonary fibrosis by activating fibroblast in an endoplasmic reticulum stress‐dependent manner

doi: 10.1111/jcmm.16524

Figure Lengend Snippet: Inhibition of ER stress attenuates SiO 2 ‐Exo‐induced lung fibroblast proliferation and migration (A‐B) CCK‐8 assay was used to evaluate the viability of HFL1 cells (A) and NIH‐3T3 cells (B) incubated with SiO 2 ‐Exos ± 4‐PBA. Two‐way ANOVA; * P < .05, ** P < .01, *** P < .001, n.s: not significant. (C‐D) The migration of HFL1 cells (C) and NIH‐3T3 cells (D) was assessed by wound closure assay. Scale bar: 100 μm. Student's t test; * P < .05, ** P < .01, *** P < .001, n.s: not significant

Article Snippet: The mouse macrophage cell line RAW264.7, mouse embryonic fibroblast cell line NIH‐3T3, human monocyte leukaemia cell line THP‐1 and human embryonic fibroblast cell line HFL1 were purchased from the China Center for Type Culture Collection (CCTCC).

Techniques: Inhibition, Migration, CCK-8 Assay, Incubation, Wound Closure Assay