Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: THSG alleviates cerebral ischemia/reperfusion injury via the GluN2B-CaMKII-ERK1/2 pathway.

doi: 10.1016/j.phymed.2024.155595

Figure Lengend Snippet: Fig. 5. The pro-survival effect of GluN2B and its transcriptional upregulation by THSG following OGD/R. (A) The relative mRNA levels of GluN2A and GluN2B in neurons after OGD/R by real-time PCR (n = 3, from independent experiments). (B) Cell viability in different concentrations of PEAQX and Ro 25–6981 by CCK-8 (n = 3, from independent experiments). (C) Effect of PEAQX (0.27 μM) and Ro 25–6981(0.9 nM) on autophagic flux and the number of autophagosomes (yellow column) and autolysosomes in neurons (red column), Bar: 40 μm (n = 3). (D) The relative mRNA levels of GluN2B after THSG treatment. The cells were treated with 1 μM THSG. Statistical comparisons were performed with one-way ANOVA. ###p < 0.001 vs. control group. *p < 0.05, ***p < 0.001 vs. model group. Data are shown as the mean ± SEM.

Article Snippet: The neurons were treated with THSG (0.1, 0.3, 1, 3, and 10 μM), the GluN2A antagonist PEAQX (0.027, 0.27, and 2.7 μM; T4171, Topscience), Ro 25–6981 (0.09, 0.9, and 9 nM), or the mitophagy inhibitor 3-methyladenine (3-MA, 5 mM; T1879, Topscience).

Techniques: Real-time Polymerase Chain Reaction, CCK-8 Assay, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: THSG alleviates cerebral ischemia/reperfusion injury via the GluN2B-CaMKII-ERK1/2 pathway.

doi: 10.1016/j.phymed.2024.155595

Figure Lengend Snippet: Fig. 6. Upregulation of GluN2B protein by THSG and the reversion of Ro 25–6981. (A-B) Expression of GluN2B in penumbra or in primary neurons after CI/R by WB and the Semi-quantitation (n = 3). (C) Representative TTC-stained brain slices and brain infarct volumes of each group shown in bar graph (n = 6). (D) neurological deficit scores of each group shown in bar graph (n = 6). (E) HE and Nissl staining showing neuron injury in rats after CI/R (blue arrows indicate nuclear pyknosis, green arrows indicate neuronal edema and black arrows indicate Nissl bodies). Bar: 2000 μm (upper panels), 80 μm, (lower panels). (F) Expression of Bcl-2, Bax, Caspase-3 in penumbra by WB and the semi-quantitation (n = 3). (G) Cell viability by CCK8 (n = 3). (H) Expression of Bcl-2, Bax, Caspase-3 in neurons after OGD/R by WB and the semi-quantitation (n = 3). The rats were treated with 10 mg/kg Ro 25–6981 in C-F and the cells were administrated with 1 μM THSG and 0.9 nM Ro 25–6981 in B, G-H. Statistical comparisons were performed with one-way ANOVA. ##p < 0.01, ###p < 0.001 vs. control group. *p < 0.05, **p < 0.01, ***p < 0.001 vs. model group. &p < 0.05, &&p < 0.01, &&&p < 0.001 vs. THSG group. Data are shown as the mean ± SEM.

Article Snippet: The neurons were treated with THSG (0.1, 0.3, 1, 3, and 10 μM), the GluN2A antagonist PEAQX (0.027, 0.27, and 2.7 μM; T4171, Topscience), Ro 25–6981 (0.09, 0.9, and 9 nM), or the mitophagy inhibitor 3-methyladenine (3-MA, 5 mM; T1879, Topscience).

Techniques: Expressing, Quantitation Assay, Staining, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: THSG alleviates cerebral ischemia/reperfusion injury via the GluN2B-CaMKII-ERK1/2 pathway.

doi: 10.1016/j.phymed.2024.155595

Figure Lengend Snippet: Fig. 7. The activation of PINK1/PARK2 pathway by THSG treatment and the reversion of Ro 25–6981. (A) PARK2 level in neurons of penumbra by IF. Bar: 100 μm. (B) Expression of PINK1, PARK2 in penumbra by WB and the semi-quantitation (n = 3). (C) The colocalization of PARK2 with mitochondria in neurons after OGD/R. Bar: 10 μm. (D) Expression of PINK1, PARK2 in neurons after OGD/R by WB and the semi-quantitation (n = 3). The rats were treated with 10 mg/kg Ro 25–6981 in A-B and the cells were administrated with 1 μM THSG and 0.9 nM Ro 25–6981 in C-D. Statistical comparisons were performed with one-way ANOVA. #p < 0.05, ###p < 0.001 vs. control group. *p < 0.05, **p < 0.01, ***p < 0.001 vs. model group. &&p < 0.01, &&&p < 0.001 vs. THSG group. Data are shown as the mean ± SEM.

Article Snippet: The neurons were treated with THSG (0.1, 0.3, 1, 3, and 10 μM), the GluN2A antagonist PEAQX (0.027, 0.27, and 2.7 μM; T4171, Topscience), Ro 25–6981 (0.09, 0.9, and 9 nM), or the mitophagy inhibitor 3-methyladenine (3-MA, 5 mM; T1879, Topscience).

Techniques: Activation Assay, Expressing, Quantitation Assay, Control

Journal: Phytomedicine : international journal of phytotherapy and phytopharmacology

Article Title: THSG alleviates cerebral ischemia/reperfusion injury via the GluN2B-CaMKII-ERK1/2 pathway.

doi: 10.1016/j.phymed.2024.155595

Figure Lengend Snippet: Fig. 9. The upregulation of CaMKII and ERK1/2 phosphorylation by THSG and the reversion of Ro 25–6981. (A) Expression of ERK1/2, P-ERK1/2, CaMKII and P- CaMKII in penumbra by WB and the semi-quantitation (n = 3). (B) P-ERK1/2 level in neurons after OGD/R by IF, Bar: 20 μm. (C) Expression of ERK1/2, P-ERK1/2, CaMKII and P-CaMKII in neurons after OGD/R by WB and the semi-quantitation (n = 3). The rats were treated with 10 mg/kg Ro 25–6981 in A, and the cells were administrated with 1 μM THSG and 0.9 nM Ro 25–6981 in B-C. Statistical comparisons were performed with one-way ANOVA. ###p < 0.001 vs. control group. *p < 0.05, ***p < 0.001 vs. model group. &p < 0.05, &&p < 0.01, &&&p < 0.001 vs. THSG group. Data are shown as the mean ± SEM.

Article Snippet: The neurons were treated with THSG (0.1, 0.3, 1, 3, and 10 μM), the GluN2A antagonist PEAQX (0.027, 0.27, and 2.7 μM; T4171, Topscience), Ro 25–6981 (0.09, 0.9, and 9 nM), or the mitophagy inhibitor 3-methyladenine (3-MA, 5 mM; T1879, Topscience).

Techniques: Phospho-proteomics, Expressing, Quantitation Assay, Control

Journal: Molecular Medicine Reports

Article Title: NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury

doi: 10.3892/mmr.2015.4501

Figure Lengend Snippet: Immunohistochemical staining for NR2B in the subventricular zone at different time points (2, 6, 24 and 48 h) after hypoxic-ischemic injury in the control, sham and hypoxic-ischemia groups.

Article Snippet: Selective non-competitive NMDAR antagonist MK-801 (M107), NR2A antagonist NVP-AAM077 (P1999), and NR2B antagonist Ro25-6981 (R7150) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Medicine Reports

Article Title: NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury

doi: 10.3892/mmr.2015.4501

Figure Lengend Snippet: Comparison of the number of NR2B-positive cells in the sub-ventricular zone (SVZ) at different time points (2, 6, 24 and 48 h) after hypoxic-ischemic injury in the control (labeled 'normal') group, sham group and hypoxic-ischemia (labeled 'model') group. Data are presented as mean ± standard deviation. Positive cell counts in five 100×100 μ m 2 areas in the SVZ of three serially sectioned brains at each tested time point. ** P<0.01.

Article Snippet: Selective non-competitive NMDAR antagonist MK-801 (M107), NR2A antagonist NVP-AAM077 (P1999), and NR2B antagonist Ro25-6981 (R7150) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Labeling, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury

doi: 10.3892/mmr.2015.4501

Figure Lengend Snippet: Immunohistochemical staining for Nestin in the subventricular zone at different time points (2, 6, 24 and 48 h) after hypoxic-ischemic injury in the control, sham and hypoxic-ischemia groups. MK, MK-801; Ro, Ro25-6981; NVP, NVP-AAM077.

Article Snippet: Selective non-competitive NMDAR antagonist MK-801 (M107), NR2A antagonist NVP-AAM077 (P1999), and NR2B antagonist Ro25-6981 (R7150) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Medicine Reports

Article Title: NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury

doi: 10.3892/mmr.2015.4501

Figure Lengend Snippet: Comparison of the number of integrated optical density (IOD) value of Nestin-positive cells in the subventricular zone (SVZ) at different time points (2, 6, 24 and 48 h) after hypoxic-ischemic injury in the sham group, hypoxic-ischemia (labeled 'model') group, hypoxic-ischemia+treatment with MK-801 group, hypoxic-ischemia+treatment with Ro25-6981 group, and hypoxic-ischemia+treatment with NVP-AAM077 group. Data are presented as mean ± standard deviation. Positive cell counts in five 100×100 μ m 2 areas in the SVZ of three serially sectioned brains at each tested time point. * P<0.05 and ** P<0.01 vs. hypoxic-ischemia group. # P<0.05 and ## P<0.01 vs. hypoxic-ischemia+treatment with MK group.

Article Snippet: Selective non-competitive NMDAR antagonist MK-801 (M107), NR2A antagonist NVP-AAM077 (P1999), and NR2B antagonist Ro25-6981 (R7150) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Labeling, Standard Deviation

Journal: Molecular Medicine Reports

Article Title: NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury

doi: 10.3892/mmr.2015.4501

Figure Lengend Snippet: Immunohistochemical staining for DCX in the subventricular zone at different time points (2, 6, 24 and 48 h) after hypoxic-ischemic injury in the control, sham and hypoxic-ischemia groups. DCX, Nestin and doublecortin; MK, MK-801; Ro, Ro25-6981; NVP, NVP-AAM077.

Article Snippet: Selective non-competitive NMDAR antagonist MK-801 (M107), NR2A antagonist NVP-AAM077 (P1999), and NR2B antagonist Ro25-6981 (R7150) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Immunohistochemical staining, Staining

Journal: Molecular Medicine Reports

Article Title: NMDA receptors promote neurogenesis in the neonatal rat subventricular zone following hypoxic-ischemic injury

doi: 10.3892/mmr.2015.4501

Figure Lengend Snippet: Comparison of the number of DCX-positive cells in the subventricular zone (SVZ) at different time points (2, 6, 24 and 48 h) after hypoxic-ischemic in the sham group, hypoxic-ischemia (labeled 'model') group, hypoxic-ischemia+treatment with MK-801 group, hypoxic-ischemia+treatment with Ro25-6981 group, and hypoxic-ischemia+treatment with NVP-AAM077 group. Data are presented as mean ± standard deviation. Positive cell counts in five 100×100 μ m 2 areas in the SVZ of three serially sectioned brains at each tested time point. * P<0.05, ** P<0.01, and △ P<0.001 vs. hypoxic-ischemia group. #P<0.05, ## P<0.01, and ★ P<0.001 vs. hypoxic-ischemia+treatment with MK group.

Article Snippet: Selective non-competitive NMDAR antagonist MK-801 (M107), NR2A antagonist NVP-AAM077 (P1999), and NR2B antagonist Ro25-6981 (R7150) were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Techniques: Labeling, Standard Deviation

Journal: Molecular Psychiatry

Article Title: Monoacylglycerol lipase inhibitors produce pro- or antidepressant responses via hippocampal CA1 GABAergic synapses

doi: 10.1038/mp.2016.22

Figure Lengend Snippet: JZL184 induces in vivo long-term depression (LTD) at CA3-CA1 synapses. ( a–g ) Plots of normalized field excitatory postsynaptic potential (fEPSP) slopes in anesthetized mice ( a, d–g ) or rats ( b, c ) show that an administration of JZL184 ( a, d–g ) or 2-AG ( b, c ) at 0 min elicits LTD lasting for >2 h in wild-type ( a, d–g ), Glu- CB 1 R -KO and GABA- CB 1 R -KO mice ( e ), but not in GFAP- CB 1 R -KO mice ( d ), which is blocked by Ro25,6981 ( f ) and Tat-GluR2 ( g ), as actinomycin-D blocks the late-phase expression of 2-AG-elicited LTD ( c ). Representative fEPSP traces before (1) and after (2) vehicle, JZL184, or 2-AG administration are shown below each plot. ( h ) Histogram summarizes the average percent change of fEPSP slope before (1) and after (2) vehicle, JZL184 or 2-AG administration as depicted in a–g . All summary graphs show means±s.e.m.s; n =numbers of animals recorded in each group ( a–g ). * P <0.05 and ** P <0.01 vs control, t test ( b–d, f, g ) or LSD post-hoc test after one-way ANOVA ( a: F 2,9 =31.904, P <0.01; e: F 2,6 =27.102, P =0.892).

Article Snippet: After recording of baseline field excitatory postsynaptic potentials (fEPSPs) for 20 min, animals received different treatment before fEPSP recording for 120 min: (I) JZL184 (5 or 20 mg kg −1 , i.p.), fluoxetine (10mg kg −1 , i.p.) or vehicle; (II) Ro25,6981 (Sigma; 6 mg kg −1 , i.p.; dissolved in physiological saline) or vehicle 10 min before JZL184; (III) Tat-GluR2 or Tat-GluR2s (1.5 μmol kg −1 , i.p.) 2 h before JZL184; (IV) intra-CA1 iontophoretic ejection (−40 nA for 30 sec) of 2-AG (50 μg ml −1 ) or vehicle; (V) actinomycin-D (Sigma; 72 μg/12 μl, intracerebroventricular injection; dissolved in 10% Tween-80, 20% dimethyl sulfoxide and 70% physiological saline) or vehicle 2 h before a 2-AG ejection.

Techniques: In Vivo, Expressing