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