Journal: Acta pharmaceutica Sinica. B

Article Title: Honokiol alleviated neurodegeneration by reducing oxidative stress and improving mitochondrial function in mutant SOD1 cellular and mouse models of amyotrophic lateral sclerosis.

doi: 10.1016/j.apsb.2022.07.019

Figure Lengend Snippet: Figure 5 Honokiol activated the NRF2 anti-oxidant pathway in the SOD1-G93A cells. (A) Luminescence intensity of HEK293-ARE reporter cells after treatment with indicated concentrations of honokiol for 6, 12 and 24 h (n Z 3); (B) Representative images of NRF2 nuclear trans- localization after 24 h honokiol treatment. Scale bars, 8 mm; (C) Western blot analysis and quantification of NRF2 in nuclear and cytoplasmic cell lysates after 24 h honokiol treatment (n Z 4); (D) RT-qPCR analysis of transcription levels of Hmox1, Nqo1, Gclm, Gclc, Gss and Gsr after 12 h honokiol treatment (n Z 3e7); (E) Western blot analysis and quantification of HO-1, NQO1, GCLC, GCLM, GSS and GSR in total cell lysates after 24 h honokiol treatment (n Z 3e6). One-way ANOVA followed by post hoc LSD test was used for the comparison among three or more groups (SPSS 16.0). Data are presented as the mean SEM. #P < 0.05, ###P < 0.001 versus pEGFP control group; *P < 0.05, **P < 0.01, ***P < 0.001 versus SOD1-G93A model group.

Article Snippet: Membranes were blocked by the 5% skimmed milk solution for 2 h at room temperature and then incubated at 4 C overnight with antibodies against BCL2 (26539- 1-AP, 1:1000), BAX (50599-2-Ig, 1:2000), cytochrome c (10993- 1-AP, 1:2000), ATM (27156-1-AP, 1:1000), NRF2 (16396-1-AP, 1:1000), GCLC (12601-1-AP, 1:2000), GCLM (14241-1-AP, 1:2000), OPA1 (27733-1-AP, 1:1000), MFN2 (12186-1-AP, 1:2000), FIS1 (10956-1-AP, 1:1000), p62 (18420-1-AP, 1:1000), GFAP (26825-1-AP, 1:2000), HSP90 (13171-1-AP, 1:2000), GAPDH (60004-1-Ig, 1:5000) from Proteintech (Rosemont, IL, USA); GSR (sc-133245, 1:500), GSS (sc-166882, 1:500), VDAC1 (sc-390996, 1:500), PGC-1a (sc-518025, 1:500), NRF1 (sc515360, 1:500), TFAM (sc-166965, 1:500) from Santa Cruz Biotechnology (Dallas, TX, USA); PINK1 (BC100-494S, 1:500) from Novus biological (Littleton, CO, USA); 4-HNE (ab46545, 1:1000), HO-1 (ab68477, 1:2000), NQO1 (ab80588, 1:1000) from Abcam (Cambridge, UK).

Techniques: Western Blot, Quantitative RT-PCR, Comparison, Control

Journal: Acta pharmaceutica Sinica. B

Article Title: Honokiol alleviated neurodegeneration by reducing oxidative stress and improving mitochondrial function in mutant SOD1 cellular and mouse models of amyotrophic lateral sclerosis.

doi: 10.1016/j.apsb.2022.07.019

Figure Lengend Snippet: Figure 9 Honokiol activated the NRF2eGSH pathway and improved mitochondrial function in the spinal cord and gastrocnemius muscle of SOD1-G93A mice. (A) and (D) Western blot analysis and quantification of HO-1, NQO1, GCLC, GCLM, GSS and GSR in the spinal cord (n Z 3e4); (B) and (E) Western blot analysis and quantification of PGC-1a, NRF1 and TFAM in the spinal cord (n Z 4); (C) and (F) Western blot analysis and quantification of OPA1, MFN2 and FIS1 in the spinal cord (n Z 4); (G) and (J) Western blot analysis and quantification analysis of HO-1, NQO1, GCLC, GCLM, GSS and GSR in the gastrocnemius muscle (n Z 4); (H) and (K) Western blot analysis and quantification of PGC-1a, NRF1 and TFAM in the gastrocnemius muscle (n Z 4e6); (I) and (L) Western blot analysis and quantification of OPA1, MFN2 and FIS1 in the gastrocnemius muscle (n Z 4). One-way ANOVA followed by post hoc LSD test was used for the comparisons among three or more groups (SPSS 16.0). Data are presented as the mean SEM (n Z 3e6, repeated independent experiments). #P < 0.05, ##P < 0.01 versus WT control group; *P < 0.05, **P < 0.01 versus SOD1-G93A model group.

Article Snippet: Membranes were blocked by the 5% skimmed milk solution for 2 h at room temperature and then incubated at 4 C overnight with antibodies against BCL2 (26539- 1-AP, 1:1000), BAX (50599-2-Ig, 1:2000), cytochrome c (10993- 1-AP, 1:2000), ATM (27156-1-AP, 1:1000), NRF2 (16396-1-AP, 1:1000), GCLC (12601-1-AP, 1:2000), GCLM (14241-1-AP, 1:2000), OPA1 (27733-1-AP, 1:1000), MFN2 (12186-1-AP, 1:2000), FIS1 (10956-1-AP, 1:1000), p62 (18420-1-AP, 1:1000), GFAP (26825-1-AP, 1:2000), HSP90 (13171-1-AP, 1:2000), GAPDH (60004-1-Ig, 1:5000) from Proteintech (Rosemont, IL, USA); GSR (sc-133245, 1:500), GSS (sc-166882, 1:500), VDAC1 (sc-390996, 1:500), PGC-1a (sc-518025, 1:500), NRF1 (sc515360, 1:500), TFAM (sc-166965, 1:500) from Santa Cruz Biotechnology (Dallas, TX, USA); PINK1 (BC100-494S, 1:500) from Novus biological (Littleton, CO, USA); 4-HNE (ab46545, 1:1000), HO-1 (ab68477, 1:2000), NQO1 (ab80588, 1:1000) from Abcam (Cambridge, UK).

Techniques: Western Blot, Control

Journal: Journal of Neurotrauma

Article Title: Advanced Age and Neurotrauma Diminish Glutathione and Impair Antioxidant Defense after Spinal Cord Injury

doi: 10.1089/neu.2022.0010

Figure Lengend Snippet: Glutathione synthesis and recycling. Glutathione is a recyclable tripeptide synthesized in two reactions. First, Glutamate-Cysteine Ligase ligates glutamate and cysteine, followed by glutathione synthetase, which adds glycine to finish the tripeptide structure. Two reduced glutathiones (GSH) are used as substrates for glutathione peroxidase (GPx) during the sequestration of reactive oxygen species (ROS). This oxidized glutathione (GSSG) is a dimer formed as a disulfide bridge that can be reduced by glutathione reductase (GR) back into two independent glutathione compounds using nicotinamide adenine dinucleotide phosphate (NADPH) as a substrate. GCLC, glutamate cysteine ligase catalytic domain.

Article Snippet: Blots were imaged using fluorescent development (Odyssey CLx; LI-COR Biosciences) and probed sequentially against GR (56 kD; 1:2,000; ab124995; Abcam Co.) and GCLC (70 kD; 1:1,000; H00002729-M01; Novus Biologicals), followed by GPx-1 (22 kD; 1:1,000; VPA00488; Bio-Rad) and GSH Synthetase (56 kD; 1:2,000; VMA00364; Bio-Rad), and finally against GAPDH (36 kD; 1:5,000; ab9484; Abcam Co.).

Techniques: Synthesized