Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 3 | Am80 attenuated neurological deficits, neuronal apoptosis, and BBB permeability at 24 h after SAH. Moreover, Am80 reduced hippocampus injury 14 days after SAH. (A, B) The modified Garcia and beam balance scores of each group. n = 6 per group. (C) Quantification of Evans blue extravasation at 24 h after SAH. (D) Representative images of TUNEL staining (red) in neurons (NeuN, green). (E) Quantitative analyses of TUNEL-positive cells. Scale bar = 100 mm. n=3 per group. (F) Representative images of Nissl staining in the CA1, CA3, and DG regions. (G) Upper middle indicates the hippocampus division and location of staining. Quantitative analyses of Nissl staining. n=3 per group. Scale bar = 100 mm. *P < 0.05 vs sham group; **P < 0.01 vs sham group; #P < 0.05 vs SAH+vehicle group; ##P < 0.05 vs SAH+vehicle group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: Permeability, TUNEL Assay, Staining

Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 4 | Am80 attenuated neuronal apoptosis in vitro SAH model. (A) Illustration of the co-culture system. (B) Quantitative analyses of cell viability as detected by CCK8 assays. n=3 per group. (C–G) Western blotting and quantitative analyses of the expression of Bcl-xl, Bcl-2, Bax, and cleaved-caspase 3 of SH-SY5Y after co-cultivation. n=3 per group. (H) Immunofluorescence staining of TUNEL (red) and SH-SY5Y (green) in an in vitro co-culture system. Scale bar=50 mm. n=3 per group. (I) Representative flow cytometry images of SH-SY5Y cells, the necroptosis is represented by the ratio of PI +/annexin V+. n=3 per group. *P < 0.05 vs control group; **P < 0.01 vs control group; #P < 0.05 vs Hb+vehicle group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: In Vitro, Co-Culture Assay, Western Blot, Expressing, Staining, TUNEL Assay, Cytometry, Control

Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 5 | Am80 attenuated the excessive activation of microglia after SAH. (A, B) Immunofluorescence staining and quantitative analysis of activated microglia (Iba-1). Scale bar = 100 mm. n = 3 per group. (C, D) Western blotting and quantitative analyses of Iba-1 expression. n=6 per group. (E–I) Representative images of activated-microglia (green) used for the measurement of morphological parameters and quantification analysis, including area, lacunarity, fractal dimension and perimeter in the basal cortex. Scale bar = 50 mm. n=6 per group. *P < 0.05 vs sham group; **P < 0.01 vs sham group; #P < 0.05 vs SAH+vehicle group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: Activation Assay, Staining, Western Blot, Expressing

Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 6 | Am80 promoted M1-to-M2 phenotypic polarization of microglia in an in vitro model of SAH. (A–C) Western blotting and quantitative analyses Arg-1 and iNOS expression of microglia in an in vitro co-culture system. n=3 per group. (D–F) Immunofluorescence staining of Arg-1/iNOS and microglia in an in vitro co-culture system and quantitative analyses of Arg-1/iNOS positive cells. Scale bar=50 mm. n = 3 per group. *P < 0.05 vs control group; **P < 0.01 vs control group; #P < 0.05 vs Hb+vehicle group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: In Vitro, Western Blot, Expressing, Co-Culture Assay, Staining, Control

Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 7 | AGN196996 abolished the protective effects of Am80. (A) Schematic diagram of experimental design. (B, C) The neurological score were evaluated at 24 h after SAH. n = 6 per group. (D–L) Western blotting and quantitative analyses of the expression of RARa, Mafb, Msr1, p-Akt, p-NF-kB, IL-6, IL-1b and TNF-a. n = 6 per group. (M) Immunofluorescence staining and quantitative analyses of TNF-a-positive cells field ratio. Scale bar=50 mm. n = 3 per group. *P < 0.05 vs sham group; #P < 0.05 vs SAH+vehicle group; @P < 0.05 vs SAH+Am80 group; &P < 0.05 vs SAH+AGN196996+Am80 group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: Western Blot, Expressing, Staining

Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 8 | Msr1 siRNA and LY294002 abolished the anti-neuroinflammation effects of Am80. (A, B) The neurological score were evaluated at 24 h after SAH. n = 6 per group. (C–J) Western blotting and quantitative analyses of the expression of Mafb, Msr1, p-Akt, p-NF-kB, IL-6, IL-1b and TNF-a. n = 6 per group. (K) Immunofluorescence staining and quantitative analyses of TNF-a-positive cells field ratio. Scale bar = 50 mm. n=3 per group. *P < 0.05 vs SAH group; #P < 0.05 vs SAH+Scr siRNA+Am 80 group; &P < 0.05 vs SAH+DMSO+Am 80 group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: Western Blot, Expressing, Staining

Journal: Frontiers in immunology

Article Title: Activation of RARα Receptor Attenuates Neuroinflammation After SAH via Promoting M1-to-M2 Phenotypic Polarization of Microglia and Regulating Mafb/Msr1/PI3K-Akt/NF-κB Pathway.

doi: 10.3389/fimmu.2022.839796

Figure Lengend Snippet: FIGURE 9 | Am80 administration suppressed the nuclear transcription of NF-kB p65 in vitro model of SAH. (A–D) Western blotting and quantitative analyses of the expression of p-IKB-a and NF-kB p65 in microglia after co-cultivation. (E–H) Western blotting and quantitative analysis of the expression of IL-6, IL-1b and TNF-a in microglia after co-cultivation. n = 3 per group. *P < 0.05 vs control group; **P < 0.01 vs control group; #P < 0.05 vs Hb+vehicle group.

Article Snippet: Am80 (Tocris, Ellisville, USA) was dissolved in carboxymethyl cellulose solution and administered by intraperitoneal (i.p.) injection 1 h after SAH.

Techniques: In Vitro, Western Blot, Expressing, Control