Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Delivery of immunoglobulin G antibodies to the rat nervous system following intranasal administration: Distribution, dose-response, and mechanisms of delivery

doi: 10.1016/j.jconrel.2018.08.006

Figure Lengend Snippet: Tissue concentrations a (pM) of [ 125 I]-IgG following intranasal saline (control) vs MMP-9 (100 nM) pre-treatment.

Article Snippet: Matrix metalloproteinase-9 (MMP-9): activated rat MMP-9 (Sino Biological; catalog number: 80049-R08H) was diluted in saline and administered intranasally, as described previously [ 25 ].

Techniques:

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Delivery of immunoglobulin G antibodies to the rat nervous system following intranasal administration: Distribution, dose-response, and mechanisms of delivery

doi: 10.1016/j.jconrel.2018.08.006

Figure Lengend Snippet: Fold-change in [125I]-IgG concentrations reaching the brain following matrix metalloproteinase-9 (MMP-9) pre-administration (100 nM) compared to saline (control) pre-administration via the nasal route. Fold-change in [125I]-IgG brain concentrations derived from data presented as mean ± S.E.M. in Table 4. Grey line indicates fold-change with respect to the intranasal saline pre-administration control data. Abbreviations: IN – intranasal; pt. – point; ACA – anterior cerebral artery; MCA – middle cerebral artery; Bas. – Basilar artery; Vert. – Vertebral arteries; PVS – perivascular space; cerv. – cervical.

Article Snippet: Matrix metalloproteinase-9 (MMP-9): activated rat MMP-9 (Sino Biological; catalog number: 80049-R08H) was diluted in saline and administered intranasally, as described previously [ 25 ].

Techniques: Derivative Assay

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Delivery of immunoglobulin G antibodies to the rat nervous system following intranasal administration: Distribution, dose-response, and mechanisms of delivery

doi: 10.1016/j.jconrel.2018.08.006

Figure Lengend Snippet: Fluorescence imaging of AF488-IgG distribution in the brain following intranasal delivery. Intranasal delivery of AF488-IgG following 100 nM matrix metalloproteinase-9 (MMP-9) intranasal pre-treatment resulted in AF488-IgG signal in the olfactory bulb nerve layer (A-D) as well as along perivascular spaces of cerebral blood vessels (B-D). AF488-IgG signal in the perivascular compartment of the anterior cerebral circulation was observed at (E) the rhinal fissure and (F) at the longitudinal fissure between the two brain hemispheres. (G, H) Perivascular AF488-IgG signal in the rhinal fissure was localized outside the smooth muscle layer of vessels identified as putative arteries based on morphology. A-H: laser scanning confocal microscopy. Vascular endothelial cells were labeled with an anti-RECA-1 antibody; astrocytes were labeled with an anti-GFAP antibody; smooth muscle cells were labeled with an anti-α-SMA antibody; cell nuclei were labeled using DAPI. Representative images from n = 3 rats. Abbreviations: gl – glomerular layer of the olfactory bulb; onl – olfactory nerve layer of the olfactory bulb; pvs – perivascular space; RECA-1 – rat endothelial cell antigen-1; GFAP – glial fibrillary acidic protein; α-SMA – alpha smooth muscle actin; DAPI: 4,6-diamidino-2-phenylindole.

Article Snippet: Matrix metalloproteinase-9 (MMP-9): activated rat MMP-9 (Sino Biological; catalog number: 80049-R08H) was diluted in saline and administered intranasally, as described previously [ 25 ].

Techniques: Fluorescence, Imaging, Confocal Microscopy, Labeling

Journal: eLife

Article Title: Homeostatic regulation of perisynaptic matrix metalloproteinase 9 (MMP9) activity in the amblyopic visual cortex

doi: 10.7554/elife.52503

Figure Lengend Snippet: Figure 5. DE lowers the threshold for light-induced activation of MMP2/9. (A) Dark chamber with an imaging window allows maintenance of visual deprivation during two photon live imaging of MMP2/9 biomarker. Left drawing: Top view of a subject wearing a custom aluminum headpost (1 cm diameter) magnetically held to an o-ring in the blackout ceiling of the dark camber (inset; 3 mm diameter magnets APEX magnets; magnetic field generation around V1,<20 gauss). The headpost is secured to a stereotax. The cannula for biomarker delivery is adjacent to theimaging window margin. Right drawing: Side view of a subject in the dark chamber. The headpost is magnetically attached to the o-ring opening of the blackout ceiling (magnet locations, yellow arrows). (B) In vitro emission spectrum of MMP2/9 biomarker A580 (2 ng/ml) incubated with activated rat recombinant MMP9 (rrMMP9, 100 ng). (C) Inset: Experimental timeline. Adult (>P90) WT mice received AAV-CaMKII-GFP~2 weeks before 10 d of DE. Biomarker was delivered 24 hr before imaging. Subjects received 40 s of light stimulation (1 Hz flash of 470 nm LED at 0 or 300 cd/m2). Left: Representative images of GFP (green) and biomarker (MMP, magenta) signals in V1b 10 s prior or 40 s after light stimulation at 0 or 300 cd/m2 in a DE subject. Right: Time course of raw fluorescent intensities (pixel) and DF/F of MMP biomarker (top) and co-localized GFP (bottom) within the single ROI denoted by yellow circle, from 10 s before (10) to 40 s after (+40) light stimulation of 0 or 300 cd/ m2 in a DE subject. (D) Summary data: Time course of DF/F of MMP biomarker from 10 s to +40 s of light stimulation of 0 or 300 cd/m2 in DE subjects. DF/F of MMP biomarker was stable in absence of visual stimulation (0 cd/m2) and increased over time in response to 300 cd/m2 light stimulation (mean ± SEM; Repeated measure ANOVA, F(1, 22), *p<0.001; n = 12 puncta from three subjects each). (E) Biomarker DF/F +40 s relative to 0 s as a function of DE (0, hr, 18 hr or 10 d) and light intensity (0, 300, or 150,000 cd/m2). Moderate intensity light did not induce a change in biomarker fluorescence in the absence of DE (blue line, p=0.49, Student’s T-test; n = 11, 10 puncta for 0 and 300 cd/m2, respectively). Following 18 hr of dark adaptation, a significant increase in biomarker fluorescence was observed in response to high, but not moderate intensity light (black line, One-way ANOVA, F(2, Figure 5 continued on next page

Article Snippet: Fluorescence spectrum measurement of MMP2/9 biomarker The small peptide MMP2/9 biomarker (A580) was dissolved in PBS (200 ng/ml) and incubated with 100 ng of activated rat recombinant MMP9 (rrMMP9, R and D Systems, according to activation protocol provided by the supplier) at 37 ̊ overnight with light protection.

Techniques: Activation Assay, Imaging, Biomarker Discovery, In Vitro, Incubation, Recombinant, Fluorescence

Journal: Molecules

Article Title: Ginsenoside Rb1 from Panax notoginseng Suppressed TNF-α-Induced Matrix Metalloproteinase-9 via the Suppression of Double-Strand RNA-Dependent Protein Kinase (PKR)/NF-κB Pathway

doi: 10.3390/molecules27228050

Figure Lengend Snippet: The anti-MMP-9 activity of the active fraction of PNG-3 and Ginsenoside Rb1 (GsRb1) from Panax notoginseng and their mechanism of action.

Article Snippet: The recombinant rat TNF-α and rat total MMP-9 ELISA kit were bought from R&D systems, Minneapolis, MN, USA.

Techniques: Activity Assay

Journal: Molecules

Article Title: Ginsenoside Rb1 from Panax notoginseng Suppressed TNF-α-Induced Matrix Metalloproteinase-9 via the Suppression of Double-Strand RNA-Dependent Protein Kinase (PKR)/NF-κB Pathway

doi: 10.3390/molecules27228050

Figure Lengend Snippet: Isolation and identification of the bioactive compound Ginsenoside Rb1 (GsRb1). ( A ) The extraction scheme of GsRb1 from Panax notoginseng (PNG). PNG powder was first sonicated in 10% ethanol (EtOH) at room temperature. The dry extract was further successively dissolved in methanol (MeOH), and the residue was dissolved in water (H 2 O). The extracts were tested for their inhibitory effects on MMP-9 activity on TNF-α-induced H9c2 cells. The most potent extract, PNG-3, was further separated using column chromatography and reverse-phased HPLC until a single bioactive molecule was isolated. * Represents the fractions with inhibitory effect on TNF-α-induced MMP-9 activity. Other extraction methods (Method 1, 3 and 4) are also summarized. ( B ) Chemical structure of Ginsenoside Rb1.

Article Snippet: The recombinant rat TNF-α and rat total MMP-9 ELISA kit were bought from R&D systems, Minneapolis, MN, USA.

Techniques: Isolation, Sonication, Activity Assay, Column Chromatography

Journal: Molecules

Article Title: Ginsenoside Rb1 from Panax notoginseng Suppressed TNF-α-Induced Matrix Metalloproteinase-9 via the Suppression of Double-Strand RNA-Dependent Protein Kinase (PKR)/NF-κB Pathway

doi: 10.3390/molecules27228050

Figure Lengend Snippet: Effect of F5 subfractions (F5-1 to F5-6) and Compound P on TNF-α-induced MMP-9 activity using gelatin zymography. ( A ) H9c2 cells were seeded at 5 × 10 4 cells/mL. After being pretreated with the six subfractions of F5 (50 μg/mL) separately for 24 h, the cells were stimulated by recombinant rat TNF-α (10 ng/mL) for another 72 h. ( B ) H9c2 cells were pretreated with compound P (25 μg/mL, 50 μg/mL) and F5-4 (25 μg/mL, 50 μg/mL) separately and then stimulated by recombinant rat TNF-α (10 ng/mL) for another 72 h. Results are shown as mean ±SD, N = 4. The fold induction of MMP-9 activity in H9c2 cells was normalized with that of the TNF-α-only treated cells. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001; # p < 0.05 vs. TNF-α+Compound P (25 μg/mL); ^ p < 0.05 vs. TNF-α+Compound P (50 μg/mL).

Article Snippet: The recombinant rat TNF-α and rat total MMP-9 ELISA kit were bought from R&D systems, Minneapolis, MN, USA.

Techniques: Activity Assay, Zymography, Recombinant

Journal: Molecules

Article Title: Ginsenoside Rb1 from Panax notoginseng Suppressed TNF-α-Induced Matrix Metalloproteinase-9 via the Suppression of Double-Strand RNA-Dependent Protein Kinase (PKR)/NF-κB Pathway

doi: 10.3390/molecules27228050

Figure Lengend Snippet: Effects of Ginsenoside Rb1 (GsRb1) on MMP-9 production in H9c2 and HepG-2 cells. ( A ) Effect of GsRb1 on TNF-α-induced MMP-9 gene expression in H9c2 cells. Cells (5 × 10 4 ) were pretreated with different concentrations of GsRb1 (1, 10, 25, 50 μg/mL) for 24 h and were stimulated by recombinant rat TNF-α (10 ng/mL) for another 24 h. ( B , C ) Effect of GsRb1 on TNF-α-induced MMP-9 production and activity in H9c2 cells. ( D – F ) Effect of GsRb1 on TNF-α-induced MMP-9 gene expression, protein production, and activity in HepG-2 cells. Results are shown as mean ± SD, N = 4. The fold induction was normalized with that of the TNF-α-only treated cells. *** p < 0.001, **** p < 0.0001.

Article Snippet: The recombinant rat TNF-α and rat total MMP-9 ELISA kit were bought from R&D systems, Minneapolis, MN, USA.

Techniques: Expressing, Recombinant, Activity Assay

Journal: Molecules

Article Title: Ginsenoside Rb1 from Panax notoginseng Suppressed TNF-α-Induced Matrix Metalloproteinase-9 via the Suppression of Double-Strand RNA-Dependent Protein Kinase (PKR)/NF-κB Pathway

doi: 10.3390/molecules27228050

Figure Lengend Snippet: PNG-3 and Ginsenoside Rb1 (GsRb1) suppressed TNF-α-induced MMP-9 via PKR/NF-κB in H9c2 cells. ( A ) 2-AP suppressed TNF-α-induced MMP-9 gene expression in H9c2 cell. ( B ) 2-AP suppressed TNF-α-induced MMP-9 activity dose-dependently in H9c2 cell. ( C – E ) PNG-3 suppressed the phosphorylation of PKR and eIF-2α and preserved the expression of IκB. ( F ) GsRb1 suppressed the phosphorylation of PKR, eIF-2α and the nucleic expression of NF-κB p65, as well as preserved the expression of IκB. Results are shown as mean ± SD, N = 4 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. control or TNF-α only.

Article Snippet: The recombinant rat TNF-α and rat total MMP-9 ELISA kit were bought from R&D systems, Minneapolis, MN, USA.

Techniques: Expressing, Activity Assay

Journal: Molecules

Article Title: Ginsenoside Rb1 from Panax notoginseng Suppressed TNF-α-Induced Matrix Metalloproteinase-9 via the Suppression of Double-Strand RNA-Dependent Protein Kinase (PKR)/NF-κB Pathway

doi: 10.3390/molecules27228050

Figure Lengend Snippet: PNG-3 and Ginsenoside Rb1 (GsRb1) suppressed TNF-α/PMA-induced MMP-9 via PKR/NF-κB in HepG-2 cells. ( A ) 2-AP suppressed TNF-α-induced MMP-9 gene expression in HepG-2 cell. ( B ) 2-AP and PNG-3 suppressed PMA-induced MMP-9 activity in HepG-2 cell. ( C – E ) PNG-3 extract suppressed TNF-α-induced phosphorylation of PKR and eIF-2α and preserved the expression of IκB. ( F ) GsRb1 suppressed TNF-α-induced phosphorylation of PKR, eIF-2α and the nucleic expression NF-κB p65, as well as preserved the expression of IκB. Results are shown as mean ± SD, N = 4 in each group. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. control or TNF-α only.

Article Snippet: The recombinant rat TNF-α and rat total MMP-9 ELISA kit were bought from R&D systems, Minneapolis, MN, USA.

Techniques: Expressing, Activity Assay

Journal: Journal of Immunology Research

Article Title: Effect of SIS3 on Extracellular Matrix Remodeling and Repair in a Lipopolysaccharide-Induced ARDS Rat Model

doi: 10.1155/2020/6644687

Figure Lengend Snippet: The rat primer sequences used in this study.

Article Snippet: RAGE, TGF- β 1, MMP2, and MMP9 concentrations in the rat sera and BALF were determined using a rat RAGE ELISA kit (EK0971 from Boster Biological Technology, China), a rat TGF- β 1 ELISA kit (MB100B, R&D Systems), a rat MMP2 ELISA kit (MMP200, R&D Systems), and a rat MMP9 ELISA kit (CSB-E08008r Cusabio, Wuhan, China), respectively, following the manufacturer's protocols.

Techniques:

Journal: Journal of Immunology Research

Article Title: Effect of SIS3 on Extracellular Matrix Remodeling and Repair in a Lipopolysaccharide-Induced ARDS Rat Model

doi: 10.1155/2020/6644687

Figure Lengend Snippet: SIS3 inhibits mRNA expression of RAGE, TGF- β 1, MMP2, and MMP9 in lung homogenates of ARDS rats. The mRNA levels of RAGE, TGF- β 1, MMP2, and MMP9 were determined using real-time PCR and were standardized to β -actin. The results suggested that SIS3 inhibited the increase of (a) RAGE, (b) TGF- β 1, (c) MMP2, and (d) MMP9 in LPS-induced lung homogenates of ARDS rats, while no effect was seen upon pretreatment with PBS. The data are presented as the means ± SD ( n = 10). ★ P < 0.05 vs. CTL; # P < 0.05 vs. ARDS.

Article Snippet: RAGE, TGF- β 1, MMP2, and MMP9 concentrations in the rat sera and BALF were determined using a rat RAGE ELISA kit (EK0971 from Boster Biological Technology, China), a rat TGF- β 1 ELISA kit (MB100B, R&D Systems), a rat MMP2 ELISA kit (MMP200, R&D Systems), and a rat MMP9 ELISA kit (CSB-E08008r Cusabio, Wuhan, China), respectively, following the manufacturer's protocols.

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Journal of Immunology Research

Article Title: Effect of SIS3 on Extracellular Matrix Remodeling and Repair in a Lipopolysaccharide-Induced ARDS Rat Model

doi: 10.1155/2020/6644687

Figure Lengend Snippet: SIS3 pretreatment decreases the protein expression and localization of RAGE, TGF- β 1, MMP2, and MMP9 in ARDS rats. Immunohistochemical staining of lung tissue sections showed that RAGE, TGF- β 1, MMP2, and MMP9 were expressed in the bronchial smooth muscle, airways, and alveolar epithelial cells of rats. The expression levels of RAGE, TGF- β 1, MMP2, and MMP9 in the ECM of ARDS rats induced by LPS were significantly higher than those of the control group. RAGE, TGF- β 1, MMP2, and MMP9 were reduced by pretreatment with SIS3, and no effect was found upon pretreatment with PBS. The micrographs were magnified at 400x. The red triangles indicate infiltrated leukocytes.

Article Snippet: RAGE, TGF- β 1, MMP2, and MMP9 concentrations in the rat sera and BALF were determined using a rat RAGE ELISA kit (EK0971 from Boster Biological Technology, China), a rat TGF- β 1 ELISA kit (MB100B, R&D Systems), a rat MMP2 ELISA kit (MMP200, R&D Systems), and a rat MMP9 ELISA kit (CSB-E08008r Cusabio, Wuhan, China), respectively, following the manufacturer's protocols.

Techniques: Expressing, Immunohistochemical staining, Staining, Control

Journal: Journal of Immunology Research

Article Title: Effect of SIS3 on Extracellular Matrix Remodeling and Repair in a Lipopolysaccharide-Induced ARDS Rat Model

doi: 10.1155/2020/6644687

Figure Lengend Snippet: SIS3 inhibited the protein expression of RAGE, TGF- β 1, MMP2, and MMP9 proteins in lung homogenates acquired from ARDS rats. The expression levels of RAGE, TGF- β 1, MMP2, and MMP9 proteins in lung tissue homogenates, sera, and BALF of ARDS rats and were determined using western blotting analysis at 24 h after LPS intervention. The results of western blotting showed that SIS3 inhibited LPS-induced (a) RAGE, (b) TGF- β 1, (c) MMP2, and (d) and MMP9 protein expression in the lung homogenates. The data are presented as the means ± SD ( n = 10). ★ P < 0.05 vs. CTL; # P < 0.05 vs. ARDS.

Article Snippet: RAGE, TGF- β 1, MMP2, and MMP9 concentrations in the rat sera and BALF were determined using a rat RAGE ELISA kit (EK0971 from Boster Biological Technology, China), a rat TGF- β 1 ELISA kit (MB100B, R&D Systems), a rat MMP2 ELISA kit (MMP200, R&D Systems), and a rat MMP9 ELISA kit (CSB-E08008r Cusabio, Wuhan, China), respectively, following the manufacturer's protocols.

Techniques: Expressing, Western Blot

Journal: Journal of Immunology Research

Article Title: Effect of SIS3 on Extracellular Matrix Remodeling and Repair in a Lipopolysaccharide-Induced ARDS Rat Model

doi: 10.1155/2020/6644687

Figure Lengend Snippet: SIS3 pretreatment prevented the reduction of the expression of RAGE, TGF- β 1, MMP2, and MMP9 in LPS-induced ARDS. The effects of SIS3 on the protein expression levels of (a) RAGE, (b) TGF- β 1, (c) MMP2, and (d) MMP9 in the BALF and sera of the ARDS rats were determined using ELISA. The results demonstrated that the levels of RAGE, TGF- β 1, MMP2, and MMP9 proteins were significantly higher in the ARDS group than in the control group, while the levels of RAGE, TGF- β 1, MMP2, and MMP9 in the SIS3 group were lower than those in the ARDS group. The protein levels of RAGE, TGF- β 1, MMP2, and MMP9 in LPS-administered pretreatment with PBS were not different from those of the ARDS group. The data are presented as the means ± SD of three independent experiments in triplicate ( n = 10). ★ P < 0.05 vs. CTL; # P < 0.05 vs. ARDS.

Article Snippet: RAGE, TGF- β 1, MMP2, and MMP9 concentrations in the rat sera and BALF were determined using a rat RAGE ELISA kit (EK0971 from Boster Biological Technology, China), a rat TGF- β 1 ELISA kit (MB100B, R&D Systems), a rat MMP2 ELISA kit (MMP200, R&D Systems), and a rat MMP9 ELISA kit (CSB-E08008r Cusabio, Wuhan, China), respectively, following the manufacturer's protocols.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Control