Journal: Frontiers in Pharmacology

Article Title: Recombinant Annexin A2 Administration Improves Neurological Outcomes After Traumatic Brain Injury in Mice

doi: 10.3389/fphar.2021.708469

Figure Lengend Snippet: rA2 administration reduces BBB leakage after TBI in mice. (A) Dose-range (0.75, 1.0, and 1.5 mg/kg) effects of rA2 administration treated at 2 h in BBB permeability examined at 24 h after TBI. (B) The therapeutic time window of rA2 administration in BBB permeability reduction examined at 24 h after TBI, treated at 2 h, or 4 h, or 6 h after TBI, respectively. Data are expressed as mean ± SEM, n = 8 mice per group, * p < 0.05 vs. BSA treated ipsilateral hemisphere. (C) Representative gel images of western blotting and quantitative analysis for ZO-1, VE-Cadherin, Occludin, Claudin-5 expression in the ipsilateral hemispheres collected at 24 h after TBI. β-actin was used as the equal loading control. Data are expressed as mean ± SEM, n = 4 mice per group, * p < 0.05 vs. Sham, # p < 0.05 vs. TBI+vehicle (1 mg/kg BSA).

Article Snippet: HBMEC were grown in endothelial complete medium (EndoGRO-MV-VEGF Complete Culture Media Kit, EMD Millipore, SCME003) to about 100% confluence and then insulted with hypoxia (with a modular chamber (Billups-Rothenberg)) perfused with 90% N 2 /5% CO 2 /5% H 2 for 30 min at 37°C) for 24 h. For rA2 or rA2 plus KG-501 (2-naphthol-AS-E-phosphate, Selleckchem, S8409) co-treatment, HBMEC were cultured in endothelial medium containing 1% FBS to 80% confluence and then treated with rA2 plus DMSO or KG501 for 24 h. Tube formation and scratch migration assays were used as indirect in vitro assays to assess angiogenesis as we previously described ( ).

Techniques: Permeability, Western Blot, Expressing, Control

Journal: Frontiers in Pharmacology

Article Title: Recombinant Annexin A2 Administration Improves Neurological Outcomes After Traumatic Brain Injury in Mice

doi: 10.3389/fphar.2021.708469

Figure Lengend Snippet: rA2 administration augments mRNA levels of pro-angiogenic factors in the isolated cerebral microvascular fragments at 7 days after TBI in mice. Gene expression levels of pro-angiogenic factors in the brain microvessels were measured at 7 days after TBI by RT-qPCR. Quantitative analysis of mRNA levels of VEGF (A) , Ang1 (B) , Tie2 (C) , eNOS (D) , IGF1 (E) , and BDNF (F) . Data are expressed as mean ± SEM; n = 4 mice per group, * p < 0.05 compared to Sham, # p < 0.05 compared to TBI+vehicle (1 mg/kg BSA).

Article Snippet: HBMEC were grown in endothelial complete medium (EndoGRO-MV-VEGF Complete Culture Media Kit, EMD Millipore, SCME003) to about 100% confluence and then insulted with hypoxia (with a modular chamber (Billups-Rothenberg)) perfused with 90% N 2 /5% CO 2 /5% H 2 for 30 min at 37°C) for 24 h. For rA2 or rA2 plus KG-501 (2-naphthol-AS-E-phosphate, Selleckchem, S8409) co-treatment, HBMEC were cultured in endothelial medium containing 1% FBS to 80% confluence and then treated with rA2 plus DMSO or KG501 for 24 h. Tube formation and scratch migration assays were used as indirect in vitro assays to assess angiogenesis as we previously described ( ).

Techniques: Isolation, Gene Expression, Quantitative RT-PCR

Journal: Frontiers in Pharmacology

Article Title: Recombinant Annexin A2 Administration Improves Neurological Outcomes After Traumatic Brain Injury in Mice

doi: 10.3389/fphar.2021.708469

Figure Lengend Snippet: rA2 induces CREB phosphorylation, VEGF, and BDNF in HBEMC. (A) Representative MAPKs profile array and quantification in HBMEC treated with PBS or rA2 (2 μg/ml) for 24 h, n = 3 experiments. (B) Representative western blotting images of p-CREB, CREB, VEGF, BDNF, and β-actin and quantification for p-CREB, VEGF, BDNF in HBMEC treated with dose-dependent rA2 for 24 h n = 3 experiments. (C) Representative gel images and quantification of western blotting for VEGF, BDNF in HBMEC treated with rA2 (2 μg/ml) with or without KG-501 (10 μM) for 24 h. (D) Viability of cultured HBMEC after rA2 with/without KG501 treatment. n = 3 experiments. Data are expressed as mean ± SEM, * p < 0.05 compared to PBS, # p < 0.05 compared to rA2.

Article Snippet: HBMEC were grown in endothelial complete medium (EndoGRO-MV-VEGF Complete Culture Media Kit, EMD Millipore, SCME003) to about 100% confluence and then insulted with hypoxia (with a modular chamber (Billups-Rothenberg)) perfused with 90% N 2 /5% CO 2 /5% H 2 for 30 min at 37°C) for 24 h. For rA2 or rA2 plus KG-501 (2-naphthol-AS-E-phosphate, Selleckchem, S8409) co-treatment, HBMEC were cultured in endothelial medium containing 1% FBS to 80% confluence and then treated with rA2 plus DMSO or KG501 for 24 h. Tube formation and scratch migration assays were used as indirect in vitro assays to assess angiogenesis as we previously described ( ).

Techniques: Phospho-proteomics, Western Blot, Cell Culture

Journal: Frontiers in Pharmacology

Article Title: Recombinant Annexin A2 Administration Improves Neurological Outcomes After Traumatic Brain Injury in Mice

doi: 10.3389/fphar.2021.708469

Figure Lengend Snippet: rA2 administration enhances cerebral angiogenesis and function vessel density after TBI in mice. Cerebral angiogenesis was determined at 14 days after TBI by bromodeoxyuridine (BrdU) and CD31 double immunostaining method, and functional vessel density was examined at 28 days by lectin perfused brain vessels analysis. (A) . Representative images and quantification of CD31 + /BrdU + cells (indicated by white arrows, cell number per filed) at hippocampus (Hippo) and peri-lesion cortex (Peri) areas of TBI mouse brains. (B) . Representative images of lectin and quantification of ipsilateral/contralateral vessel density (%) in the hippocampus (Hippo), SVZ, and peri-lesion cortex (Peri) areas of TBI mouse brains. Scale bar = 50 μm. Data are expressed as mean ± SEM, n = 6 mice per group. * p < 0.05 vs. TBI+vehicle (1 mg/kg BSA).

Article Snippet: HBMEC were grown in endothelial complete medium (EndoGRO-MV-VEGF Complete Culture Media Kit, EMD Millipore, SCME003) to about 100% confluence and then insulted with hypoxia (with a modular chamber (Billups-Rothenberg)) perfused with 90% N 2 /5% CO 2 /5% H 2 for 30 min at 37°C) for 24 h. For rA2 or rA2 plus KG-501 (2-naphthol-AS-E-phosphate, Selleckchem, S8409) co-treatment, HBMEC were cultured in endothelial medium containing 1% FBS to 80% confluence and then treated with rA2 plus DMSO or KG501 for 24 h. Tube formation and scratch migration assays were used as indirect in vitro assays to assess angiogenesis as we previously described ( ).

Techniques: Double Immunostaining, Functional Assay

Journal: Frontiers in Pharmacology

Article Title: Recombinant Annexin A2 Administration Improves Neurological Outcomes After Traumatic Brain Injury in Mice

doi: 10.3389/fphar.2021.708469

Figure Lengend Snippet: rA2 administration improves neurological outcomes and reduces brain tissue loss after TBI in mice. (A) . Modified neurological severity score (NSS). (B) Rotor rod test. * p < 0.05 compared to TBI+vehicle (1 mg/kg BSA) at the same time point. (C) Y-maze test was performed at 21 days after TBI. n = 14 mice per group. * p < 0.05 compared to Sham, # p < 0.05 compared to TBI+vehicle (1 mg/kg BSA). (D) Representative MAP2 staining and quantification of lesion volume. The dashed line indicates the border of the brain tissue loss. Scar bar, 1 mm * p < 0.05 compared to TBI+vehicle (1 mg/kg BSA). Data are expressed as mean ± SEM; n = 14 mice per group.

Article Snippet: HBMEC were grown in endothelial complete medium (EndoGRO-MV-VEGF Complete Culture Media Kit, EMD Millipore, SCME003) to about 100% confluence and then insulted with hypoxia (with a modular chamber (Billups-Rothenberg)) perfused with 90% N 2 /5% CO 2 /5% H 2 for 30 min at 37°C) for 24 h. For rA2 or rA2 plus KG-501 (2-naphthol-AS-E-phosphate, Selleckchem, S8409) co-treatment, HBMEC were cultured in endothelial medium containing 1% FBS to 80% confluence and then treated with rA2 plus DMSO or KG501 for 24 h. Tube formation and scratch migration assays were used as indirect in vitro assays to assess angiogenesis as we previously described ( ).

Techniques: Modification, Staining

Journal: International journal of molecular medicine

Article Title: Resveratrol attenuates pulmonary embolism associated cardiac injury by suppressing activation of the inflammasome via the MALAT1‑miR‑22‑3p signaling pathway.

doi: 10.3892/ijmm.2019.4358

Figure Lengend Snippet: Figure 1. RS increased promoter efficiency of MALAT1. (A) Fragment of MALAT1 promoter was inserted into immediately upstream of luciferase gene. (B) Luciferase activity in HL‑1 cells transfected with 1 and 5 µm RS compared with the negative controls. *P<0.05 vs. the negative controls. (C) Luciferase activity in AC16 cells transfected with 1 and 5 µm RS compared with the negative controls. *P<0.05 vs. the negative controls. RS, resveratrol; MALAT1, metastasis associated lung adenocarcinoma tran- script 1; NC, negative control.

Article Snippet: There are three groups of rats (n=8 per group): Sham groups as the control, animal model of PE group and PE group treated with resveratrol (RS; 30 mg/kg; Santa Cruz Biotechnology, Inc.) solution.

Techniques: Luciferase, Activity Assay, Transfection, Negative Control

Journal: International journal of molecular medicine

Article Title: Resveratrol attenuates pulmonary embolism associated cardiac injury by suppressing activation of the inflammasome via the MALAT1‑miR‑22‑3p signaling pathway.

doi: 10.3892/ijmm.2019.4358

Figure Lengend Snippet: Figure 5. MALAT1, miR‑22‑3p and NLRP3 are differently expressed in different groups. (A) MALAT1 level in PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. P<0.05 vs. the sham group. (B) miR‑22‑3p level in PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. (C) NLRP3 level in PE associated cardiac injury group, PE asso- ciated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. miR, microRNA; PE, pulmonary embolism; RS, resveratrol.

Article Snippet: There are three groups of rats (n=8 per group): Sham groups as the control, animal model of PE group and PE group treated with resveratrol (RS; 30 mg/kg; Santa Cruz Biotechnology, Inc.) solution.

Techniques:

Journal: International journal of molecular medicine

Article Title: Resveratrol attenuates pulmonary embolism associated cardiac injury by suppressing activation of the inflammasome via the MALAT1‑miR‑22‑3p signaling pathway.

doi: 10.3892/ijmm.2019.4358

Figure Lengend Snippet: Figure 6. NLRP3, ASC and Caspase‑1 are differently expressed in different groups. (A) NLRP3, ASC and Caspase‑1 levels in PE associated cardiac injury group, PE associated cardiac injury plus RS group, and sham group. *P<0.05 vs. the sham group. (B) NLRP3 level in PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. (C) ASC level in the PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. (D) Caspase‑1 level in the PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. PE, pulmonary embolism; RS, resveratrol.

Article Snippet: There are three groups of rats (n=8 per group): Sham groups as the control, animal model of PE group and PE group treated with resveratrol (RS; 30 mg/kg; Santa Cruz Biotechnology, Inc.) solution.

Techniques:

Journal: International journal of molecular medicine

Article Title: Resveratrol attenuates pulmonary embolism associated cardiac injury by suppressing activation of the inflammasome via the MALAT1‑miR‑22‑3p signaling pathway.

doi: 10.3892/ijmm.2019.4358

Figure Lengend Snippet: Figure 7. Differential levels of inflammatory factors including IL‑1β and IL‑18 in various groups. (A) IL‑1β level in the PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. (B) IL‑18 level in PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group. *P<0.05 vs. the sham group. IL, interleukin; PE, pulmonary embolism; RS, resveratrol.

Article Snippet: There are three groups of rats (n=8 per group): Sham groups as the control, animal model of PE group and PE group treated with resveratrol (RS; 30 mg/kg; Santa Cruz Biotechnology, Inc.) solution.

Techniques:

Journal: International journal of molecular medicine

Article Title: Resveratrol attenuates pulmonary embolism associated cardiac injury by suppressing activation of the inflammasome via the MALAT1‑miR‑22‑3p signaling pathway.

doi: 10.3892/ijmm.2019.4358

Figure Lengend Snippet: Figure 8. NLRP3 protein in PE associated cardiac injury group, PE associated cardiac injury plus RS group and sham group (magnification, x40). PE, pulmonary embolism; RS, resveratrol.

Article Snippet: There are three groups of rats (n=8 per group): Sham groups as the control, animal model of PE group and PE group treated with resveratrol (RS; 30 mg/kg; Santa Cruz Biotechnology, Inc.) solution.

Techniques: