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