Journal: Drug Design, Development and Therapy

Article Title: Integrated Serum Pharmacochemistry, Network Pharmacology, and Transcriptomics Reveal the Mechanisms and Active Constituents of Qingfei Huoxue Decoction Against Bleomycin-Induced Pulmonary Fibrosis

doi: 10.2147/DDDT.S592770

Figure Lengend Snippet: Effects of QFHXD on histopathology and pro-fibrotic markers in BLM-induced PF mice. ( A ) Dynamic changes in body weight. ( B ) The Kaplan-Meier survival curve of each group. Compared with the model group, Treatment with different doses of QFHXD markedly boosted the survival rate of PF mice. ( C ) The representative images of lung sections stained with H&E, Masson’s trichrome, and immunohistochemical staining for α-SMA and FN proteins (×200 magnification, scale bar: 100 μm, n = 6). ( D ) Ashcroft score for grading PF severity (n = 6). Quantification of immunohistochemical results of α-SMA ( E ) and FN ( F ) expression (n = 6). The relative mRNA levels of Fn1 ( G ) and Col1a1 ( H ) in lung samples of each group (n = 5). QFHXD administration effectively mitigated fibrotic lesions, inhibited collagen deposition, and decreased the expression of pro-fibrotic markers. Serum levels of TGF-β1 ( I ), CCL2 ( J ), and KL-6 ( K ) were measured by ELISA (n = 8). Data are displayed as the mean ± SD. ## P < 0.01 versus the control group; * P < 0.05, and ** P < 0.01 versus the model group. Model, BLM-induced PF model; QFHXD-L, QFHXD low dose (4.78 g/kg); QFHXD-H, QFHXD high dose (9.56 g/kg).

Article Snippet: BLM (99.03%, HY-17565), human TGF-β1 protein (HY-P7118), and SB-431542 (HY-10431) were obtained from MedChemExpress.

Techniques: Histopathology, Staining, Immunohistochemical staining, Expressing, Enzyme-linked Immunosorbent Assay, Control

Journal: Drug Design, Development and Therapy

Article Title: Integrated Serum Pharmacochemistry, Network Pharmacology, and Transcriptomics Reveal the Mechanisms and Active Constituents of Qingfei Huoxue Decoction Against Bleomycin-Induced Pulmonary Fibrosis

doi: 10.2147/DDDT.S592770

Figure Lengend Snippet: Experimental validation of inflammatory targets regulated by QFHXD in BLM-induced PF. ( A and B ) The relative mRNA levels of key inflammatory targets obtained from network pharmacology and RNA-Seq analysis, including Tlr2, Tlr4, Rela, Nfkb1, Il1b, Il6, Tnf, Tgfb1, Ccl2, Spp1, Mmp9, Mmp13, and Mmp14 (n = 5). The concentrations of TLR2 ( C ), TLR4 ( D ), NF-κB ( E ), IL-1β ( F ), IL-6 ( G ), TNF-α ( H ), TGF-β1 ( I ), CCL2 ( J ), and SPP1 ( K ) proteins were investigated by ELISA (n = 6). ( L ) The typical images of immunohistochemical staining of MMP9 protein in each group (×200 magnification, scale bar: 100 μm, n = 6). Black arrows indicated MMP9-positive cells. ( M ) Quantification of MMP9-positive cells (n = 6). Compared with the untreated PF model group, QFHXD intervention significantly suppressed the mRNA and protein expression of pro-inflammatory mediators in fibrotic lung samples. RT-qPCR results are expressed as the mean ± SEM. Data of ELISA and immunohistochemical staining are displayed as the mean ± SD. ## P < 0.01 versus the control group; * P < 0.05, and ** P < 0.01 versus the model group. Model, BLM-induced PF model; QFHXD-L, QFHXD low dose (4.78 g/kg); QFHXD-H, QFHXD high dose (9.56 g/kg).

Article Snippet: BLM (99.03%, HY-17565), human TGF-β1 protein (HY-P7118), and SB-431542 (HY-10431) were obtained from MedChemExpress.

Techniques: Biomarker Discovery, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Expressing, Quantitative RT-PCR, Control

Journal: Science Advances

Article Title: Control of airway basal stem cell–mediated lung repair by TGF-β signaling

doi: 10.1126/sciadv.adz1519

Figure Lengend Snippet: ( A ) Schematic diagram of iBMP7-BCs transplantation into BLM-damaged lungs of immune-deficient NOD-SCID mice. ( B ) Immunostaining detected pSmad2 and pSmad3 in lung tissue at different times following BLM injury. DAPI: nuclear counterstain. ( C ) pSmad2 + and pSmad3 + cells were quantified by ImageJ ( n = 6 technical replicates from three independent mice, one-way ANOVA with Dunnett’s test). ( D ) Time-series direct observations of iBMP7-BCs transplantation using a stereomicroscope. ( E ) Immunostaining for HuLamin and GFP in lung sections after iBMP7-BCs transplantation. HuLamin: lamin A/C, a human-specific nuclear antigen. DAPI: nuclear counterstain. ( F ) The relative GFP intensity was quantified by ImageJ ( n = 3 mice per group). ( G ) Immunofluorescence of BMP7-Flag and GFP expression at 7 days after transplantation (14 dpi) of iBMP7-BCs into BLM-damaged lung. DAPI: nuclear counterstain. ( H ) Axial plane micro-CT images of mouse fibrosis at day 14 following treatment with saline, Ctrl-BCs, or iBMP7-BCs. Lower panels: representative reconstructed 3D images. Red: pulmonary injury area (including intrapulmonary airways); gray: normal lung tissue. Right panel: quantitative analysis of pulmonary injury volume based on CT images ( n = 3 mice per group). ( I ) Collagen deposition was assessed by hydroxyproline content in lung tissue ( n = 4 mice per group). ( J ) qPCR analysis of Col1a1 , Col3a1 , Fn1 , and Acta2 mRNA expression levels in mouse lungs 14 days after treatment with saline, Ctrl-BCs, or iBMP7-BCs ( n = 4 mice per group; two technical replicates were performed for each mouse). ( K ) Arterial blood gas analysis was performed on the arterial blood of mice treated with saline, Ctrl-BCs, or iBMP7-BCs for 14 days ( n = 4 mice per group). All data are represented as the means ± SEM. One-way ANOVA with Tukey’s test was used for (H) to (K). * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001. ns, not significant.

Article Snippet: The mouse lung was injured by intratracheally instilling with a 3 U/kg body weight of BLM (Selleckchem, Houston, US) 7 days before transplantation.

Techniques: Transplantation Assay, Immunostaining, Immunofluorescence, Expressing, Micro-CT, Saline

Journal: RSC Advances

Article Title: Structure-based design of orthosteric and allosteric CCR2 inhibitors for potential IPF therapy

doi: 10.1039/d5ra05026j

Figure Lengend Snippet: CCR2 Expression and Pathological Validation in IPF Progression. (A–C) In silico analysis based on patient data from dataset GSE70866 . (A) The violin plot depicts CCR2 gene expression levels in the normal control and IPF groups ( GSE70866 ) (* P < 0.1). (B) The survival analysis plot outlines survival probabilities over time for groups with high and low CCR2 expression. (C) The ROC curve demonstrates the diagnostic performance of CCR2 in identifying pulmonary fibrosis. (D–J) In vivo validation using a bleomycin-induced pulmonary fibrosis mouse model. (D) Representative images of H&E and Masson's trichrome staining of lung tissues 4 weeks after intratracheal instillation of BLM in mice. (E) Pathological scoring results of H&E-stained lung tissues at week 4 (** P < 0.01). (F) Analysis of collagen area in lung tissues at week 4 (*** P < 0.001). (G) Hydroxyproline content in lung tissue of bleomycin (BLM)-induced pulmonary fibrosis mice and control mice. The hydroxyproline level was significantly increased in BLM-treated mice compared to controls (*** P < 0.001). (H) RT-qPCR analysis of CCR2 expression levels in BLM-induced idiopathic pulmonary fibrosis (*** P < 0.001). (I and J) Western blot analysis of CCR2 expression levels in lung tissues of each group (** P < 0.01).

Article Snippet: Mice in the experimental group were anesthetized with 1% sodium pentobarbital (50 mg kg −1 ), and a single dose of 5 mg kg −1 bleomycin (BLM) solution (Selleck, USA) was administered intratracheally.

Techniques: Expressing, Biomarker Discovery, In Silico, Gene Expression, Control, Diagnostic Assay, In Vivo, Staining, Quantitative RT-PCR, Western Blot