Journal: Respiratory research

Article Title: Altered hypoxia-induced cellular responses and inflammatory profile in lung fibroblasts from COPD patients compared to control subjects.

doi: 10.1186/s12931-024-02907-x

Figure Lengend Snippet: Fig. 1 Effect of 4 h of hypoxia exposure on gene expression levels. Genes related to remodelling: VEGFR2 (a), 5-HTR2B (b) and Collagen7 (c); cellular stress: IRE1 (d) and c-Jun (h), oxidative stress: Nrf2 (e), HIF-1α (f) and SOD3 (g) were measured by RT-qPCR in primary distal lung fibroblasts obtained from healthy subjects (n = 7) and COPD patients (n = 7) after 4 h of exposure to normoxic (21% O2) or hypoxic (1% O2) conditions. Beta-actin and 18 S were used as housekeeping genes. The data is presented as median with interquartile range. Ordinary two-way ANOVA or RM two-way ANOVA were used for unpaired and paired comparisons and the post-hoc test Fisher’s LSD was used for statistical analysis *p < 0.05, **p < 0.01

Article Snippet: The following primary antibodies were used: HIF-1α (dilution 1:50, Biotin, ab81633), HIF-2α (dilution 1:100 Novus Biologicals, Bio-Techne, #NB100132), 5HTR2B (dilution 1:300, Aviva System Biology, OAAF02801), VEGFR2 (dilution 1:200, Bio-rad, AHP1327), VEGFR3 (dilution 1:200, Abcam, GR3217142-6) and negative control (Dako, X0903), all diluted in 1% BSA in TBS.

Techniques: Gene Expression, Quantitative RT-PCR

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: M. suaveolens extract blocked the expression of VEGF and NF-κB mRNA in lung tissue. Groups of mice were challenged with cecal ligation and puncture and treated with M. suaveolens extract 24 h later. The left lung tissues were homogenized and total RNA was extracted using TRIzol reagent and assayed by quantitative polymerase chain reaction. (A) Representative gels assessing (a) VEGF and (b) NF-κB levels are demonstrated. Lane 1, marker; lane 2, normal control group; lane 3, sham operation group; lane 4, (untreated) sepsis group and lane 5, treatment group. (B) Statistical summary of the densitometric analysis of VEGF and NF-κB mRNA expression in rats from the four groups; (a) VEGF mRNA and (b) NF-κB mRNA. Data are represented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. sham operation and normal control groups; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Real-time Polymerase Chain Reaction, Marker, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on the expression of VEGF and NF-κB65 protein in lung tissue. Groups of mice were challenged with cecal ligation and puncture and treated with M. suaveolens extract 24 h later. The expression of VEGF, NF-κβ65 and GAPDH was detected by western blotting using specific antibodies. GAPDH protein was used an internal control. (A) Representative western blot analysis demonstrated the levels of VEGF and NF-κB65 protein expression in rats from the four groups; (a) normal control group; (b) sham operation group; (c) (untreated) sepsis group and (d) treatment group. (B) Quantification of the blots by densitometric analysis of (a) VEGF and (b) NF-κB65 protein expression in rats from the four groups. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. the sham operation group and normal control group; # P<0.05 vs . (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Western Blot, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on the protein expression of VEGF and NF-κB65 in rat lungs 24 h following cecal ligation and puncture-induced acute lung injury. Immunostaining was performed on lung sections following antigen retrieval using Retrievagen. (A) Representative immunostaining revealed VEGF and NF-κβ65-positive expression in rats from the four groups: (a–c) Expression of positive VEGF in the (a) sham operation group; (b) control group; (c) treatment group); (d–f) Expression of positive NF-κB65 in the (d) sham operation group; (e) (untreated) sepsis group; (f) treatment group (magnification, ×200). (B) Quantification of the images by densitometric analysis of (a) VEGF and (b) NF-κB-positive protein expression in rats from four groups. All values are expressed as the mean ± standard deviation. ** P<0.01 vs. the sham operation group; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Expressing, Ligation, Immunostaining, Control, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Effect of M. suaveolens extract on plasma levels of VEGF, TNF-α, IL-6, IL-1β, IL-4 and IL-10 levels in plasma. Groups of mice were challenged with cecal ligation and puncture, and treated with M. suaveolens extract 24 h later. (A) VEGF, (B) TNF-α, (C) IL-6, (D) IL-1β, (E) IL-10 and (F) IL-4 levels in plasma were determined by ELISA. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. * P<0.05, ** P<0.01 vs. the sham operation group and normal control group; # P<0.05, ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B; TNF-α, tumor necrosis factor-α; IL, interleukin.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Clinical Proteomics, Ligation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Administration of M. suaveolens extract attenuated lipopolysaccharide-induced pulmonary inflammation. Groups of mice were challenged with cecal ligation and puncture, and treated with M. suaveolens extract 24 h later. (A) VEGF, (B) TNF-α, (C) IL-6, (D) IL-1β, (E) IL-10 and (F) IL-4 levels in bronchoalveolar lavage were determined by ELISA. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. ** P<0.01 vs. the sham operation group and normal control group; # P<0.05, ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B; TNF-α, tumor necrosis factor-α; IL, interleukin.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Ligation, Enzyme-linked Immunosorbent Assay, Standard Deviation, Control

Journal: Molecular Medicine Reports

Article Title: Effect of Melilotus suaveolens extract on pulmonary microvascular permeability by downregulating vascular endothelial growth factor expression in rats with sepsis

doi: 10.3892/mmr.2015.3146

Figure Lengend Snippet: Administration of M. suaveolens extract reduces CLP-induced lung permeability. Rats were treated as indicated and (A) FITC-labeled albumin in the bronchoalveolar lavage fluid, (B) water content of lung tissue and (C) W/D lung weight ratio were determined 24 h following CLP challenge. Data are presented as the mean ± standard deviation of one experiment consisting of three replicates. * P<0.05, ** P<0.01 vs. the sham operation and normal control groups; ## P<0.01 vs. (untreated) sepsis group. VEGF, vascular endothelial growth factor; NF-κB, nuclear factor kappa B; TNF-α, tumor necrosis factor-α; IL, interleukin; CLP, cecal ligation and puncture; W/D, wet/dry; FITC, fluorescein isothiocyanate.

Article Snippet: Primary antibodies used included rabbit anti-VEGF monoclonal antibody (1:400), rabbit anti-NF-κB65 monoclonal antibody (1:400) (Boster Biological Technology, Ltd) and mouse anti- GAPDH monoclonal antibody (1:400) (Santa Cruz Biotechnology, Inc).

Techniques: Permeability, Labeling, Standard Deviation, Control, Ligation

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) The expression of HIF-1α and HIF-2α in gastrointestinal vascular malformations and normal vessels. Red arrow: strongly positive; Black arrow: weakly positive; Blue arrow: negative. ( B ) Percentages of positive and negative vessels in GIVM and normal tissues. ** P < 0.01.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Expressing

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) Western blot determinations of HIF-1α and HIF-2α expression at different time points of hypoxia. * P < 0.05, ** P < 0.01 vs. 0 hour. ( B ) The effect of HIF-1α and HIF-2α overexpression on the expression of VEGF, Notch1, DLL4, and Ang2. Western blot and RT-PCR demonstrated that HIF-1α and HIF-2α overexpression increased the expression of VEGF, Notch1, DLL4, and Ang2 protein and mRNA. * P < 0.05, ** P < 0.01 vs. control. ( C ) Influence of HIF-1α and HIF-2α overexpression on angiogenesis 6 and 24 h after transfection of Lenti-HIF-1α and Lenti-HIF2α. Tube formation was enhanced 6 and 24 h after transfection. ** P < 0.01 vs. control. ( D ) Fluorescence microscope observations of subintestinal vein sprouting in normal and HIF-2α-overexpressing zebrafish. *Indicates subintestinal vascular sprouts. HIF-2α overexpression significantly increased the number of subintestinal vascular sprouts. ** P < 0.01 vs. control plasmid. ( E ) Dual luciferase reporter gene assay demonstrated that HIF-2α enhanced VEGF promoter activity. ** P < 0.01 vs. control plasmid.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Western Blot, Expressing, Over Expression, Reverse Transcription Polymerase Chain Reaction, Transfection, Fluorescence, Microscopy, Plasmid Preparation, Luciferase, Reporter Gene Assay, Activity Assay

Journal: Scientific Reports

Article Title: HIF-1α and HIF-2α induced angiogenesis in gastrointestinal vascular malformation and reversed by thalidomide

doi: 10.1038/srep27280

Figure Lengend Snippet: ( A ) Immunofluorescence indicated that HIF-1α and HIF-2α expression was down-regulated by thalidomide at different concentrations. ( B ) Western blots demonstrated that the expression of HIF-1α and HIF-2α decreased with 100 and 200 μg/ml of thalidomide. * P < 0.05, ** P < 0.01. ( C ) Western blots demonstrated that thalidomide at 200 μg/ml inhibited the expression of HIF-1α and HIF-2α in HUVECs after hypoxic treatment for 24, 36, and 48 h. * P < 0.05, ** P < 0.01. ( D ) Fluorescence microscope observations of the effect of thalidomide at different concentrations on vascular development in zebrafish with HIF-2α overexpression. ** P < 0.01 vs. HIF-2α.

Article Snippet: Proteins were transferred to a nitrocellulose membrane (Millipore, Bedford, MA, USA), which was blocked using 5% milk for 2 h, and incubated with rabbit-anti-HIF-2α, VEGF, NOTCH1, DLL4, Ang2, and β-actin antibodies (all rabbit polyclonal antibodies from KangChen Bio-tech, Shanghai, China) at 4 °C overnight.

Techniques: Immunofluorescence, Expressing, Western Blot, Fluorescence, Microscopy, Over Expression

Journal: Biomedicines

Article Title: Differential Expression of CD3, TNF-α, and VEGF Induced by Olanzapine on the Spleen of Adult Male Albino Rats and the Possible Protective Role of Vitamin C

doi: 10.3390/biomedicines7020039

Figure Lengend Snippet: Photomicrographs of immunohistochemically-stained sections of the spleen with anti- vascular endothelial growth factor (VEGF) from different studied groups. ( A ) Control group, ( B ) vitamin C group showing positive brown immunoreactivity (arrows) within the red pulp, with negatively stained white pulp. ( C ) Olanzapine-treated group showing a marked increase in immunostaining within the red pulp (arrow) and negatively stained white pulp. Note, strong brown cytoplasmic staining within endothelial cells lining the venous sinuses. ( D ) Olanzapine and vitamin C group showing moderate immunostaining within the red pulp in the endothelial cells of venous sinuses (arrow) but no immunoreactivity within the white pulp. VEGF immunostaining, ×100.

Article Snippet: Sections of all studied groups were stained with rabbit monoclonal antibody (SP7) (abcam, 16669, Cambridge, UK) against CD3 at dilution 1:100, rabbit polyclonal antibody against TNF-α (abcam, ab6671, Cambridge, UK) at 1:200, and the rabbit polyclonal antibody against VEGF at 1:500 (Gene Tex, USA).

Techniques: Staining, Control, Immunostaining

Journal: Biomedicines

Article Title: Differential Expression of CD3, TNF-α, and VEGF Induced by Olanzapine on the Spleen of Adult Male Albino Rats and the Possible Protective Role of Vitamin C

doi: 10.3390/biomedicines7020039

Figure Lengend Snippet: Area percentage of VEGF of spleens in group (III) (Olanzapine-treated group) was significantly increased compared to group (I) (control group). The Olanzapine and vitamin C group (IV) was significantly decreased as compared to group (III). The differences in area percentage of VEGF between group (I) and group (II) were not significant ( p > 0.05). Results are expressed as mean ± SD and the level of significance was set for p -values less than 0.05. **: significant increase, group (III) versus group (I) ( p < 0.001). *: significant decrease, group (IV) versus group (III) ( p < 0.001).

Article Snippet: Sections of all studied groups were stained with rabbit monoclonal antibody (SP7) (abcam, 16669, Cambridge, UK) against CD3 at dilution 1:100, rabbit polyclonal antibody against TNF-α (abcam, ab6671, Cambridge, UK) at 1:200, and the rabbit polyclonal antibody against VEGF at 1:500 (Gene Tex, USA).

Techniques: Control

Journal: Neurochemistry international

Article Title: DL-3-n-butylphthalide Induced Neuroprotection, Regenerative Repair, Functional Recovery and Psychological Benefits following Traumatic Brain Injury in Mice

doi: 10.1016/j.neuint.2017.03.017

Figure Lengend Snippet: TBI mice received acute as well as daily TBI intranasal treatments until sacrifice. Expressions of regenerative factors were measured using Western blot 21 days after TBI. A. The protein levels of BDNF, VEGF, GDNF, eNOS, CXCR4, and MMP-9. B. Quantified data from A. NBP treatments significantly enhanced the expression of BDNF, VEGF, eNOS, and MMP-9 compared to the control group. There was no significant change in GDNF and CXCR4 expressions among groups. * P<0.05 versus sham group; # P<0.05 versus TBI-saline control group by one-way ANOVA followed by Bonferroni correction; n=5 per group.

Article Snippet: The primary antibodies used and the dilutions for each were rabbit anti-cleaved caspase-3 antibody (Cell Signaling, Danvers, MA) at 1:400, rabbit anti-caspase-9 antibody (Cell Signaling) at 1: 1,000, rabbit anti-cytochrome c antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) at 1:200 or rabbit anti-AIF antibody (Cell Signaling) at 1:500, rabbit anti-TNF-α (Cell Signaling) at 1:1000, rabbit anti-IL-1β (Cell Signaling) at 1:1000, rabbit anti-BDNF (BD Bioscience, San Jose, CA) at 1:2,000, rabbit anti-VEGF (BD Bioscience) at 1,000, rabbit anti-GDNF (BD Bioscience) at 1:1,000, rat anti-eNOS (BD Bioscience) at 1:1,000, rabbit anti-CXCR4 (BD Bioscience) at 1:2,000, rabbit anti-p65 (Cell Signaling) at 1:1,000, mouse anti-IκB (Cell Signaling) at 1:1,000, mouse anti-p-IκB (Cell Signaling) at 1:1,000, rabbit anti-COX (Cell Signaling) at 1:2,500, mouse anti-actin (Sigma-Aldrich) at 1:5000, and rabbit anti-MMP-9 antibody (Millipore, Billerica, MA) at 1:2500.

Techniques: Western Blot, Expressing