Journal: International journal of molecular sciences

Article Title: Anti-Inflammatory and Neuroprotective Mechanisms of GTS-21, an α7 Nicotinic Acetylcholine Receptor Agonist, in Neuroinflammation and Parkinson's Disease Mouse Models.

doi: 10.3390/ijms23084420

Figure Lengend Snippet: Figure 1. Effect of GTS-21 on inflammatory molecules in LPS-stimulated microglial cells. (A) After pretreatment with GTS-21 for 1 h, BV2 cells or primary cultured microglia were incubated with LPS (100 ng/mL for BV2, 10 ng/mL for primary microglia). The levels of nitrite, TNF-α, IL-1β, IL-6, and TGF-β were assessed in supernatants after a 16 h incubation period (n = 3–5 per group). (B,C) BV2 cells were treated with GTS-21 for 1 h before being incubated with LPS for 6 h. Western blot analysis (B) and RT-PCR (C) were performed to determine the expression level of inflammatory molecules (n = 3–4 per group). The left panel shows representative blots/gels, whereas the right panel shows quantitative data. The data are presented as the mean ± SEM. * p < 0.05, vs. control group; ** p < 0.01, vs. control group; # p < 0.05 vs. LPS-treated group; ## p < 0.01 vs. LPS-treated group.

Article Snippet: While antibodies against phospho-p47phox were provided by Assaybiotech (Sunnyvale, CA, USA), those against TNF-α, Nrf2, HO-1, catalase, lamin A, and COX-2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Cell Culture, Incubation, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Control

Journal: International journal of molecular sciences

Article Title: Anti-Inflammatory and Neuroprotective Mechanisms of GTS-21, an α7 Nicotinic Acetylcholine Receptor Agonist, in Neuroinflammation and Parkinson's Disease Mouse Models.

doi: 10.3390/ijms23084420

Figure Lengend Snippet: Figure 5. α7 nAChR antagonists reversed the anti-inflammatory effects of GTS-21 in LPS-stimulated microglia. (A) The effect of methyllycaconitine (MLA) and α-bungarotoxin (α-BTX) on the pro- duction of NO, TNF-α, IL-6, and ROS in LPS + GTS-21-treated BV2 cells and primary microglia (n = 3–4 per group). Cells were pre-treated with MLA (1 µM) or α-BTX (0.01 µM) for 1 h, and then GTS-21 (10, 20 µM) for 1 h, followed by the treatment with LPS (100 ng/mL for BV2, 10 ng/mL for primary microglia) for 16 h. The levels of NO, TNF-α, and IL-6 released into the medium, as well as intracellular ROS, were measured. (B) The effect of MLA on NF-κB, Nrf2, CREB, and PPARγ reporter gene activities (n = 3–4 per group). BV2 cells were transfected with the reporter plasmid and then treated with MLA (1 µM) for 1 h, followed by GTS-21 (20 µM) and LPS (100 ng/mL). Cells were harvested after 6 h of LPS treatment and the reporter gene assay was carried out. (C) BV2 cells were pre-treated with MLA (1 µM) for 1 h, and then GTS-21 (10, 20 µM) for 1 h, followed by the treatment with LPS for 30 min to determine p-Akt, p-AMPK, and p-CREB levels by western blot analyses (n = 3–4 per group). The data are presented as the mean ± SEM. * p < 0.05, vs. control group; ** p < 0.01, vs. control group; # p < 0.05 vs. LPS-treated group; ## p < 0.01 vs. LPS-treated group; & p < 0.05 vs. LPS + GTS-21-treated group; && p < 0.01 vs. LPS + GTS-21-treated group.

Article Snippet: While antibodies against phospho-p47phox were provided by Assaybiotech (Sunnyvale, CA, USA), those against TNF-α, Nrf2, HO-1, catalase, lamin A, and COX-2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Transfection, Plasmid Preparation, Reporter Gene Assay, Western Blot, Control

Journal: International journal of molecular sciences

Article Title: Anti-Inflammatory and Neuroprotective Mechanisms of GTS-21, an α7 Nicotinic Acetylcholine Receptor Agonist, in Neuroinflammation and Parkinson's Disease Mouse Models.

doi: 10.3390/ijms23084420

Figure Lengend Snippet: Figure 8. Effect of GTS-21 on microglial activation, inflammatory markers, and antioxidant enzyme expression in the brains of MPTP-injected mice. (A) Iba-1-positive microglial cells in the substantia nigra and striatum (representative images). (B) Quantitative analysis was performed by measuring the number of Iba-1-positive cells (n = 4 per group, 3 sections/brain). (C–F) Protein extracts from the substantia nigra of each group were analyzed using TNF-α, iNOS, and IL-1β antibodies or Nrf2, HO-1, and NQO1 antibodies (n = 4). The figures show representative blots (C,E) and quantification data (D,F). The data are presented as the mean ± SEM. * p < 0.05, vs. control group; ** p < 0.01, vs. control group; # p < 0.05 vs. MPTP-treated group; ## p < 0.01 vs. MPTP-treated group.

Article Snippet: While antibodies against phospho-p47phox were provided by Assaybiotech (Sunnyvale, CA, USA), those against TNF-α, Nrf2, HO-1, catalase, lamin A, and COX-2 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Activation Assay, Expressing, Injection, Control