Journal: Nutrients

Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model

doi: 10.3390/nu18010055

Figure Lengend Snippet: SDA and PI protect dopaminergic neurons, reduce α-synuclein accumulation, and improve motor function in MPTP-treated mice. ( A ) Representative Nissl-stained sections of the SNc showing neuronal density loss in MPTP-treated mice, with protective effects observed following SDA and PI treatment. Quantification of Nissl-positive cells is expressed as a fold change relative to the Sham group. ( B ) TH IHC in the SNc indicates dopaminergic neuron survival. Quantified TH-positive staining is shown as a fold change versus Sham. ( C ) Immunohistochemical detection of α-synuclein in the DG, with boxed regions magnified to highlight α-synuclein-positive cells (arrowheads) in the SGZ and GCL. ( D – F ) Behavioral tests assessing motor performance: ( D ) Forelimb grip strength (grams), ( E ) Rotarod latency at a constant 20 rpm (seconds), ( F ) Wire hang endurance (seconds). Each test was conducted in triplicate with rest intervals of 5–10 min, and mean values were used for analysis. Scale bars: 200 µm ( A – C upper panels); 100 µm ( C lower panels). Data represent mean ± SEM ( n = 4 for histology, n = 9 for behavioral tests). *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.0001. DG: dentate gyrus; GCL: granule cell layer; ML: molecular layer; PBP: parabrachial pigmented nucleus of the VTA; SGZ: subgranular zone; SNc: substantia nigra pars compacta; SNr: substantia nigra pars reticulata.

Article Snippet: Parkinsonism was induced in Groups 2–5 by daily intraperitoneal (i.p.) injection of MPTP hydrochloride (30 mg/kg/day; Selleckchem, TX, USA) for four weeks.

Techniques: Staining, Immunohistochemical staining

Journal: Nutrients

Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model

doi: 10.3390/nu18010055

Figure Lengend Snippet: SDA and PI enhance hippocampal neurogenesis by promoting cell proliferation and neuronal development. ( A ) IHC for Ki67 in the SGZ of the DG shows dividing cells (arrowheads). Counting Ki67 + cells reveals significant decreases in the MPTP group, which are reversed by treatment with SDA, PI, or RES. ( B ) DCX staining highlights immature neurons within the SGZ–GCL. Boxed areas are enlarged to display representative DCX + cells (arrowheads). ( C ) WB analysis of DCX protein levels in hippocampal extracts, normalized to GAPDH, supports histological evidence of restored neurogenesis. Scale bars: 200 µm (upper panels); 100 µm (lower panels). Data represent mean ± SEM ( n = 4, 5 per group). *, p < 0.05; **, p < 0.01; ***, p < 0.005. DG: dentate gyrus; GCL: granule cell layer; ML: molecular layer; SGZ: subgranular zone.

Article Snippet: Parkinsonism was induced in Groups 2–5 by daily intraperitoneal (i.p.) injection of MPTP hydrochloride (30 mg/kg/day; Selleckchem, TX, USA) for four weeks.

Techniques: Staining

Journal: Nutrients

Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model

doi: 10.3390/nu18010055

Figure Lengend Snippet: SDA and PI restore hippocampal synaptic plasticity through PSD95 and synaptophysin expression. ( A – D ) Representative double IF images showing PSD95 (green) and synaptophysin (red) expression in hippocampal subregions (CA1, CA2/3, and DG). RES, SDA, and PI treatments reversed MPTP-induced reductions in co-localization. Quantification of co-localized PSD95 and synaptophysin is presented as intensity per mm 2 . ( E ) WB analysis of hippocampal PSD95 and synaptophysin expression normalized to GAPDH. MPTP decreased both synaptic markers, which were significantly restored by all treatments. Scale bars: 500 µm (top), 100 µm (bottom). Data represent mean ± SEM ( n = 4–5 per group). *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.0001. CA1: Cornu Ammonis 1; CA2/3: Cornu Ammonis 2/3; DG: Dentate gyrus; GCL: granule cell layer; ML: molecular layer; SO: stratum oriens; SP: stratum pyramidale; SR: stratum radiatum.

Article Snippet: Parkinsonism was induced in Groups 2–5 by daily intraperitoneal (i.p.) injection of MPTP hydrochloride (30 mg/kg/day; Selleckchem, TX, USA) for four weeks.

Techniques: Expressing

Journal: Nutrients

Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model

doi: 10.3390/nu18010055

Figure Lengend Snippet: SDA and PI decrease NOX4, MPO, and OPN expression in the hippocampus. ( A – C ) Representative double IF images showing NOX4 ( A ), MPO ( B ), and OPN ( C ) (red) in the DG. Nuclei are counterstained with DAPI (blue). Treatment with SDA and PI significantly reduced the fluorescence intensity of all three markers compared with the MPTP group. ( D ) WB analysis of NOX4, MPO, and OPN protein levels in hippocampal lysates. Densitometric quantification, normalized to GAPDH, is expressed as fold change relative to the Sham group. Quantitative analysis of WB data showing significant upregulation of NOX4, MPO, and OPN in the MPTP group and their marked reduction following treatment with RES, SDA, or PI. Both SDA and PI exhibited efficacy comparable to that of RES in attenuating oxidative/inflammatory protein expression in the hippocampus. Scale bars: 200 µm (upper panels), 100 µm (magnified). Data represent mean ± SEM ( n = 4–5 per group). *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.0001. CA1: Cornu Ammonis 1; DG: Dentate gyrus; GCL: granule cell layer; ML: molecular layer; SGZ: subgranular zone.

Article Snippet: Parkinsonism was induced in Groups 2–5 by daily intraperitoneal (i.p.) injection of MPTP hydrochloride (30 mg/kg/day; Selleckchem, TX, USA) for four weeks.

Techniques: Expressing, Fluorescence

Journal: Nutrients

Article Title: Oral Administration of Astrocyte-Targeted Natural Antioxidants Suppress NOX4-Driven Neuroinflammation and Restore Hippocampal Neurogenesis in MPTP-Induced Parkinson’s Disease Mouse Model

doi: 10.3390/nu18010055

Figure Lengend Snippet: SDA and PI restore mitochondrial integrity and inhibit apoptosis in the hippocampus. ( A ) Representative WB showing hippocampal expression of mitochondrial import and translocase proteins TIM23 and TOM20. Both SDA and PI significantly restored these mitochondrial markers that were reduced in MPTP-treated mice. ( B , C ) WB analysis of OXPHOS complex subunits I (NDUFB8), II (SDH8), III (UQCRC2), IV (MTCO1), and V (ATP5A). Quantitative densitometry demonstrates that MPTP treatment markedly suppressed each complex, whereas SDA and PI restored their levels toward those of the Sham group. ( D ) WB and densitometric analysis of 4-HNE show increased lipid peroxidation after MPTP and its significant reduction by RES, SDA, or PI treatment. ( E ) WB analysis of apoptosis-related proteins Bcl-2, BAX, and CC3. MPTP exposure increased pro-apoptotic BAX and CC3 and decreased anti-apoptotic Bcl-2; these changes were effectively reversed by SDA and PI administration. All protein levels were normalized to GAPDH and expressed as fold change relative to the Sham group. Data are presented as mean ± SEM ( n = 5 per group). Statistical comparisons were performed using one-way ANOVA followed by Dunnett’s post hoc test. *, p < 0.05; **, p < 0.01; ***, p < 0.005; ****, p < 0.0001.

Article Snippet: Parkinsonism was induced in Groups 2–5 by daily intraperitoneal (i.p.) injection of MPTP hydrochloride (30 mg/kg/day; Selleckchem, TX, USA) for four weeks.

Techniques: Expressing

Journal: Acta pharmaceutica Sinica. B

Article Title: Protocatechuic aldehyde protects cardiomycoytes against ischemic injury via regulation of nuclear pyruvate kinase M2.

doi: 10.1016/j.apsb.2021.03.021

Figure Lengend Snippet: Figure 3 PCA PKM2-dependently protects cardiomyocytes. Primary neonatal rat ventricular myocytes (NRVMs) were cultured in glucose-free DMEM under 1% O2 (OGD) for 4 h in the presence of protocatechuic aldehyde (PCA). (A) Cell survival in NRVMs exposed to OGD for 6 h (n Z 6). (B) Intracellular ROS production (n Z 6). (C) Representative images of mitochondrial permeability transition pore (mPTP) and quantification of relative fluorescence intensity (n Z 6, one of three independent experiments). Scale bar: 10 mm. (D) Mitochondrial fission was detected by Mito-Tracker Red (one of 3 independent experiments) and quantification analysis. Scale bar: 10 mm. (E) Caspase 3 activity in NRVMs (n Z 6). (F) Representative Western blots of BAX, BCL-2, caspase 3, cleaved caspase 3 (c-caspase 3) and the ratio of BAX/BCL-2 (n Z 3). (G) Representative images of TUNEL staining and quantification analysis the percentage of apoptosis cells. Scale bars, 100 mm (n Z 3). (H) Data of Annexin V/PI double staining flow cytometry; the percentage for each panel indicates the percentage of apoptotic cells and quantification of the percentage of apoptotic cells (n Z 3). Results are shown as mean SD; *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: After treatment, mPTP was detected with Mitochondrial Permeability Transition Pore Assay Kit (C2009S, Beyotime).

Techniques: Cell Culture, Permeability, Activity Assay, Western Blot, TUNEL Assay, Staining, Double Staining, Cytometry

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Experimental design. Three days before 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) injections, mice were pre-trained at the pole and beam tests by completing three trials within a day. Two days before MPTP injections, mice received an oral treatment with either vehicle [10% dimethyl sulfoxide (DMSO) and 25% Tween 20 v/v dissolved in saline 0.9% sodium chloride], FTY720 (1 mg/kg), or SEW2871 (20 mg/kg) once a day for a total of 14 days. At day 0, saline or MPTP (30 mg/kg/day) treatments were administered i.p. during 5 days. Motor abilities at the pole and beam tests were evaluated 8 days after the first MPTP injections. On day 11, all mice were sacrificed. The experimental setting involved six groups: 1) vehicle with saline treatments, n = 3; 2) FTY720 with saline treatments, n = 3; 3) SEW2871 with saline treatments, n = 3; 4) vehicle with MPTP treatments, n = 5; 5) FTY720 with MPTP treatments, n = 5; (6) SEW2871 with MPTP treatments, n = 5.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Levels of inflammatory markers are affected in the striatum. Levels of GFAP (A) , TNF-α (B) , and BDNF (C) were assessed by Western blot in the striatum of mice. The data, expressed relative to GAPDH, represent the mean of relative optical density in triplicate experiments of GFAP, TNF-α, and BDNF (expressed as a percentage of control values) ± S.E.M., n = 3–5 mice/group. ** p < 0.01, *** p < 0.001 vs . vehicle + saline; # p < 0.05, ## p < 0.01 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Western Blot, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Signs of astrogliosis and microgliosis induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) are prevented by S1P1R modulators. (A) Representative examples of Iba-1 immunoreactive microglia (panel a1 to a4; red color), GFAP-positive astrocytes (panel a5 to a8; red color) and TH-positive neurons (panel a1 to a8; green color) using epifluorescence microscope (40X objective) in the mouse subtantia nigra pars reticulate (SNr; −3.16 mm from the Bregma). Bar equals 50 µm. Fluorescence intensity of Iba-1 and GFAP were measured in the SNc (B, D) as well as in the ventral tegmental area (VTA) (C, E) . The data represent the mean of Iba-1 and GFAP relative optical density (expressed as a percentage of control values) ± S.E.M., n = 3–5 mice/group. *** p < 0.001 vs . vehicle + saline; ### p < 0.001 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Microscopy, Fluorescence, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: S1PR modulators protect against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced subtantia nigra pars compacta (SNc) cellular loss. Representative examples of tyrosine hydroxylase (TH)-immunostaining (green) using the 40X objective of the epifluorescence microscope in coronal sections (−3.16 mm from the Bregma) of mouse subtantia nigra pars reticulate (SNr). Bar equals 50 µm (A) . Stereological counts of TH- and neuronal nuclear antigen (NeuN)-positive neurons in the SNc (B, D) and ventral tegmental area (VTA) (C, E) . The data represent the mean of TH- and NeuN-positive cell numbers/mm 3 (expressed as a percentage of control values) ± S.E.M., n = 3-5 mice/group. * p < 0.05, ** p < 0.01 vs . vehicle + saline; # p < 0.05, ## p < 0.01 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Immunostaining, Microscopy, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Dopamine terminals were spared in the striatum following S1P1Rs modulators treatments. Levels of tyrosine hydroxylase (TH) (A) and dopamine transporter (DAT) (B) were determined in the mouse striatum by Western blot experiments. The data, expressed relative to GAPDH, represent the mean of relative optical density in triplicate experiments of TH and DAT (expressed as a percentage of control values) ± S.E.M., n = 3–5 mice/group. ** p < 0.01, *** p < 0.001 vs . vehicle + saline; ## p < 0.01 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Western Blot, Control, Saline

Journal: Frontiers in Pharmacology

Article Title: Sphingosine-1-Phosphate Receptors Modulators Decrease Signs of Neuroinflammation and Prevent Parkinson’s Disease Symptoms in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Mouse Model

doi: 10.3389/fphar.2020.00077

Figure Lengend Snippet: Motor behavioral outcomes. Motor abilities on the pole and beam tests were evaluated. Data represent the mean time require to perform the pole (A) and beam (B) tests and the mean number of foot-faults occurring during the beam test (C) ± S.E.M., n = 3–5 mice/group. * p < 0.05, *** p < 0.001 vs . vehicle + saline; # p < 0.05, ### p < 0.001 vs . vehicle + MPTP.

Article Snippet: FTY720 and SEW2871 [5-(4-phenyl-5-trifluoromethylthiophen-2-yl)-3-(3-trifluoromethylphenyl)-1,2,4-oxadiazole] were purchased from Cayman Chemical (Ann Arbor, MI, USA) and MPTP hydrochloride from Toronto Research Chemicals (North York, ON, CAN).

Techniques: Saline